AU1349801A - Regulation of gene expression by neuroleptic agents - Google Patents

Description

WO 01/30972 PCT/USOO/29690 REGULATION OF GENE EXPRESSION BY NEUROLEPTIC AGENTS 5 (MBHB Case No. 99,022-B) This application claims priority of U.S. Provisional Application No. 60/161,379, filed October 26, 1999 and U.S. Provisional Application No. 60/186,918, filed on March 3, 2000. Both applications are hereby incorporated by reference. 10 BACKGROUND OF THE INVENTION Schizophrenia and dopamine receptors. Midbrain dopamine neurons have been shown to play an important role in normal and diseased brain functions. For example, 15 many psychiatric disorders are associated with overactive dopaminergic activity in the meso-striatal dopamine system which refers to both the nigro-striatal dopamine pathway (neurons linking the substantia nigra to the striatum), and the meso-limbic dopamine pathway (neurons linking the ventral tegmental area to limbic regions, such as amygdala, olfactory tubercle and the nucleus accumbens, which is often considered a ventral 20 extension of the striatum). Additionally, it is known that Parkinson's Disease is caused by the degeneration of dopamine neurons of the nigro-striatal pathway. Neuroleptic (antipsychotic) drugs. Neuroleptic drugs, such as haloperidol and clozapine, are widely used in the long-term treatment of various psychiatric disorders, 25 including schizophrenia. The antipsychotic effects of neuroleptic drugs are generally attributed to blockade of D 2 receptors in the meso-limbic dopamine system (Metzler et al., Schizophrenia Bull., 2, 19-76 (1976)). The best evidence for this comes from the excellent correlation observed between the therapeutic potency of neuroleptics and their affinity for binding to the D 2 receptor (Seeman et al., Curr. Opn. Neurol. And 30 Neurosurg., 6, 602-608 (1993); Creese et al., Science, 192, 481-483 (1976); Peroutka et al., Am. J. Psych., 137, 1518-1522 (1980); Deutch, et al., Schizophren. Res., 4, 121-156 (1991); Seeman, P., Synapse 1, 133-152 (1987)). Although neuroleptic drugs have affinity for other neurotransmitter receptors in the brain, such as muscarinic WO 01/30972 PCT/USOO/29690 acetylcholine, 5-HT, alpha-adrenergic and histamine receptors, no correlation to clinical efficacy has been observed with these receptors (Peroutka et al., Am. J. Psych. (1980); Richelson et al., Eur. J. Pharm., 103, 197-204 (1984)). 5 Studies demonstrate that dopamine receptors become blocked to a level of 70% after only a few hours of neuroleptic treatment (Sedval et al., Arch. Gen. Psych., 43, 995 1006 (1986)). This blockade has been shown to lead to a compensatory increase in dopamine receptor number and supersensitivity of the unblocked receptors (Clow et al., Psychopharm., 69, 227-233 (1980); Rupniak et al., Life Sci., 32, 2289-2311 (1983); 10 Rogue et al., Eur. J. Pharm., 207, 165-169 (1991)). Furthermore, the short-term effects of dopamine antagonists on the brain are well known and include such effects as an increase in dopamine synthesis and catabolism, an increase in the firing rate of dopamine neurons resulting from the inhibition of pre-synaptic dopamine autoreceptors (Grace et al., J. Pharm. Exp. Ther., 238, 1092-1100 (1986), and a potentiation of cyclic AMP 15 formation resulting from the blockade of post-synaptic dopamine receptors (Rupniak et al., Psychopharm., 84, 519-521 (1984)). Side effects of neuroleptic drugs. In addition to their antipsychotic actions, neuroleptics can cause a series of mild to severe side effects. Some of these side-effects 20 result from the dirty nature of neuroleptic drugs, including hypotension and tachycardia, which results from alpha-adrenergic receptor blockade, and dry mouth and blurred vision, which results from the blockade of muscarinic acetylcholine receptors. The predominant and most undesirable effects that accompany neuroleptic treatment are the long-lasting motor deficits referred to as extrapyramidal side effects (Marsden et al., Psychol. Med., 25 10, 55-72 (1980)). Extrapyramidal side effects are associated with the blockade of dopamine receptors in the dorsal striatum (Moore et al., Clin. Neuropharmacol., 12, 167 184 (1989) and include such motor deficits as dystonias (muscle spasms), akathisias (motor restlessness), Parkinson's-like symptoms and Tardive Dyskinesia. Roughly 20% of patients taking antipsychotics demonstrate Parkinson's-like symptoms, the blockade of 30 dopamine D 2 receptors in the striatum being functionally equivalent to the degeneration of nigro-striatal dopamine neurons seen in Parkinson's Disease. Tardive Dyskinesia is a 2 WO 01/30972 PCT/USOO/29690 syndrome of abnormal involuntary movements that afflicts roughly 25% of patients on neuroleptic treatment (Jeste et al., Psychopharmacol., 106, 154-160 (1992)). The danger of this side effect is that it can be potentially irreversible, that is, patients can still have symptoms of Tardive Dyskinesia long after the antipsychotic has been discontinued. 5 This implicates an epigenetic component to the effects of chronic neuroleptic treatment. Interestingly, "typical" neuroleptics, such as haloperidol and fluphenazine, have a much higher propensity for causing extrapyramidal side effects than "atypical" neuroleptic drugs, such as clozapine, which rarely causes these types of effects. 10 Although clozapine differs from haloperidol in its pharmalogical profile, the specific mechanism leading to the lack of motor side effects is unclear. Since clozapine has high affinity for other neurotransmitter receptors, such as muscarinic, adrenergic and serotonin receptors, it is possible that the antipsychotic actions of clozapine are partly due to blockade of these other receptors, which may restore proper balance of the dopamine 15 input and output pathways of the basal ganglia. Genetics and genes involved in neuropsychiatric disorders. In the general population, the risk for developing a psychiatric disorder is approximately 1-2% (Maier, W., and Schwab, S., Molecular genetics of schizophrenia. Current Opinion in Psychiatry 20 11:19-25 (1998); Kendler, K.S., Twin studies of psychiatric illness: current status and future directions. Arch Gen Psychiatry 50:9095-915 (1993)). However, this risk increases to 10% or 40% if one or both parents, respectively, have the disease. Concordance in monozygotic and dizygotic twins remains only as high 40-50% (Maier and Schwab (1998)). While there is undoubtedly a genetic component to the 25 transmission of psychiatric disorders, the lack of full concordance in dizygotic twins indicates that there are other environmental factors that contribute (Maier and Schwab (1998); Kendler (1993)). A current challenge in genetic research on mental illnesses is the identification of mutations conferring susceptibility to, or genes associated with therapeutics for, such disorders. One approach addressing the latter is to identify genes 30 whose expression is altered during the process of drug treatment. 3 WO 01/30972 PCT/USOO/29690 Expression of immediate early genes resulting from acute neuroleptic treatment. Despite the immediate occupancy of dopamine receptors, neuroleptic drugs have a delayed onset of clinical action, which often can be up to several weeks. Further, as discussed above, neuroleptic drugs are characterized by their ability to cause late and 5 long-lasting motor deficits. The distinct temporal discrepancy which exists between dopamine receptor occupancy and the onset of therapeutic and extrapyramidal side effects, suggests that additional molecular changes in the brain occur downstream from dopamine receptor blockade. In an attempt to identify the downstream molecular mechanisms, studies have focused on dopamine-receptor regulation of individual target 10 genes in the striatum and nucleus accumbens. For example, several studies have demonstrated that acute treatment with antipsychotic drugs causes induction of several immediate-early genes (Nguyen et al, Proc. Natl. Acad. Sci., 89, 4270-4274 (1992); MacGibbon et al., Mol. Brain. Res. 23, 21 15 32 (1994); Robertson et al., Neuro. Sci., 46, 315-328 (1992); Dragunow et al., Neuro. Sci., 37, 287-294 (1990); MillerJ. Neurochem., 54, 1453-1455 (1990)). Some immediate early gene proteins (IEGPs) act as transcription factors by binding to specific DNA sequences and regulating gene transcription. Thus, IEGPs can link receptor-mediated signalling effects to long-term genomic activity. Recent studies have shown that 20 haloperidol, a typical neuroleptic, induces the expression c-Fos in the rat striatum and nucleus accumbens, whereas, clozapine, an atypical neuroleptic, induces c-Fos in the nucleus accumbens only (Nguyen et al., Proc. NatL. Acad. Sci. (1992); MacGibbon et al., Mol. Brain Res. (1994); Robertson et al., Neurosci. (1992)). Haloperidol has also been shown to induce expression of other IEGPs, such as FosB, JunB, JunD and Krox24, in 25 the striatum and nucleus accumbens (Rogue et al., Brain Res. Bull. 29, 469-472 (1992); Marsden et al., Psych. Med. (1980); Moore et al., Clin. Neuropharmacol. (1989)). In contrast, clozapine has been shown to induce Krox24 and JunB in the nucleus accumbens only (Nguyen et al. (1992); MacGibbon et al. (1994)). These results suggest that clozapine's lower tendency to cause extrapyramidal side effects, compared to "typical" 30 neuroleptics, may be associated with its failure to induce IEGPs in the striatum. 4 WO 01/30972 PCT/USOO/29690 The appearance of immediate early genes after acute treatment with neuroleptics likely precedes a number of other molecular changes responsible for the delayed adaptive changes that occur with drug treatment in the striatum. 5 Changes induced by chronic neuroleptic treatment. Chronic treatment with neuroleptic drugs has also been shown to cause changes in the expression of certain neuropeptides and neurotransmitter receptors. In distinct regions of the striatum, both neurotensin and enkephalin are upregulated after chronic (7 - 28 days) treatment with haloperidol, while levels of protachykinin mRNA are decreased (Merchant et al., J. 10 Pharm. Exp. Ther., 271, 460-471 (1994); Delfs et al., J. Neurochem., 63, 777-780 (1994); Angulo et al., Neurosci. Lett. 113, 217-221 (1990)). In contrast, chronic clozapine treatment results in a decrease in enkephalin mRNA levels and only small changes in the expression of neurotensin and tachykinin (Merchant et al. (1994); Mercugliano et al., Neurosci. Lett., 136, 10-15 (1992); Angulo et al. (1990)). These differences suggest that 15 neuropeptides may play a role in the motor deficits that result from treatment with typical neuroleptics. Researchers have also demonstrated the regulation of genes associated with glutaminergic neurotransmission. For example, a decrease in mRNA expression of the 20 glutamate transporter, GLT-1, was observed in the striatum after 30 days of haloperidol treatment, but not after clozapine exposure (Schneider et al., Neuroreport., 9, 133-136 (1998)). Similar treatment with haloperidol also resulted in an increase in the N-methyl D-aspartate (NMDA) receptor subunits, NR1 and NR2, whereas clozapine treatment resulted in a lesser induction (Riva et al., Mol. Brain. Res. 50, 136-142 (1997)). 25 In addition, pathological and structural changes in the striatum have been observed after chronic drug treatment. Studies using experimental animals have detected a reduction in the size and number of striatal neurons and neuronal processes, as well as decreases in striatal neuronal density following chronic treatment with haloperidol 30 (Christensen et al., Acta. Psych. Scand., 46, 14-23 (1970), Jeste et al., Psychopharm., 106, 154-160 (1992); Mahadik et al., Bio. Psych., 24, 199-217 (1988); Nielson et. al., 5 WO 01/30972 PCT/USOO/29690 Psychopharm., 59-85-89 (1978). These studies imply that neuroleptics may have a neurotoxic effect on the striatum which could account for the ensuing neuroleptic induced side effects. 5 Although the above studies have examined the expression of a few individual target genes, there has been no comprehensive study of the effects of neuroleptics on gene expression over time in the striatum and nucleus accumbens, brain regions considered to be critically involved in the actions of neuroleptic drugs. Thus, the number and identity of the genes which are differentially expressed following acute and chronic 10 treatment with neuroleptics in these tissues remains unknown. Further, there has been no comprehensive examination of the differences between the striatal mRNA expression induced by typical neuroleptics and the expression induced by atypical neuroleptics. Such a comparative study would identify the genes that regulate the antipsychotic actions of neuroleptics versus those responsible for the unwanted side effects associated with 15 these drugs. This information would advance the development of an antipsychotic therapy that would target specific actions of neuroleptic drugs or, alternatively, would selectively block proteins causing the motor side effects. In addition, a systematic characterization would allow the identification of genes 20 that contribute to neuropathologies associated with neuropsychiatric disorders, such as psychoses, bipolar disorder, and addiction-related behavior. This information can reveal pathways for the mechanism of actions of antipsychotic drugs, as well as provide insight regarding the underlying basis of psychiatric dysfunction. Specifically, the identification of potentially harmful gene products is important to identify molecules that could be 25 useful as diagnostic markers indicating neuropathology. Additionally, the identification of potentially harmful gene products is important to identify molecules that could be amenable to pharmaceutical intervention. A systematic characterization would also allow the identification of beneficial molecules that contribute to conditions of neuroprotection. Such identification of beneficial products could lead to the development of 30 pharmaceutical agents useful in the treatment of neuropsychiatric disorders. Furthermore, the identification of harmful and beneficial products may lead to new lines 6 WO 01/30972 PCT/USOO/29690 of study towards the amelioration of symptoms associated with neuropsychiatric disorders. Studies have been performed using the PCR-based Total Gene Expression 5 Analysis (TOGA) method to analyze the expression patterns of thousands of genes and compare expression patterns among time courses following clozapine drug treatment. Genes regulated by clozapine treatment were examined in haloperidol-treated animals for a comparative analysis. 10 SUMMARY OF THE INVENTION Studies have been performed using the PCR-based Total Gene Expression Analysis (TOGA) method to analyze the expression patterns of thousands of genes and compare expression patterns among time courses following clozapine drug treatment. 15 Genes regulated by clozapine treatment were examined in haloperidol-treated animals for a comparative analysis. TOGA analysis has identified several genes that are altered in their expression in response to clozapine and/or haloperidol administration in mouse brain. In particular, the TOGA system has been used to examine how gene expression in the striatum and nucleus accumbens is regulated by an atypical neuroleptic agent, such as 20 clozapine. These studies have identified proteins and genes which are regulated by the treatment of atypical drugs. Further, these studies have identified at least one gene which is differentially regulated by typical and atypical drugs. The studies have also examined the pattern of expression of neuroleptic-regulated 25 genes in various regions of the brain. Among other things, these studies are useful to determine the genes specifically associated with anti-psychotic activity versus those associated with extrapyramidal side effects, which information advances the development of improved antipsychotic therapies. The identified neuroleptic-regulated molecules are useful in therapeutic and diagnostic applications in the treatment of various 30 neuropsychiatric disorders, such as psychoses, bipolar disorder, and addiction-related behavior. Such molecules are also useful as probes as described by their size, partial 7 WO 01/30972 PCT/USOO/29690 nucleotide sequence and characteristic regulation pattern associated with neuroleptic administration. The present invention provides novel polynucleotides and the encoded 5 polypeptides. Moreover, the present invention relates to vectors, host cells, antibodies, and recombinant methods for producing the polynucleotides and the polypeptides. One embodiment of the invention provides an isolated nucleic acid molecule comprising a polynucleotide chosen from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, 10 SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO: 11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO: 49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO: 57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ 15 ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72 and SEQ ID NO:107. Also provided is an isolated nucleic acid molecule comprising a polynucleotide at least 95% identical to any one of these isolated nucleic acid molecules and an isolated nucleic acid molecule at least ten bases in length that is hybridizable to any one of these isolated nucleic acid molecules under stringent conditions. Any one of these isolated 20 nucleic acid molecules can comprise sequential nucleotide deletions from either the 5' terminus or the 3'-terminus. Further provided is a recombinant vector comprising any one of these isolated nucleic acid molecules and a recombinant host cell comprising any one of these isolated nucleic acid molecules. Also provided is the gene corresponding to the cDNA sequence of any one of these isolated nucleic acids. 25 Another embodiment of the invention provides an isolated polypeptide encoded by a polynucleotide chosen from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:1 1, SEQ ID NO:12, SEQ ID NO:13, 30 SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18. SEQ ID NO:19, SEQ ID NO: 49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID 8 WO 01/30972 PCT/USOO/29690 NO: 57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72 and SEQ ID NO:107. Also provided is an isolated nucleic acid molecule 5 encoding any of these polypeptides, an isolated nucleic acid molecule encoding a fragment of any of these polypeptides, an isolated nucleic acid molecule encoding a polypeptide epitope of any of these polypeptides, and an isolated nucleic acid encoding a species homologue of any of these polypeptides. Another embodiment of the invention provides an isolated polypeptide of SEQ ID NO: 109. Another embodiment of the 10 invention provides an isolated polypeptide of SEQ ID NO: 110. Preferably, any one of these polypeptides has biological activity. Optionally, any one of the isolated polypeptides comprises sequential amino acid deletions from either the C-terminus or the N-terminus. Further provided is a recombinant host cell that expresses any one of these isolated polypeptides. 15 Yet another embodiment of the invention comprises an isolated antibody that binds specifically to an isolated polypeptide encoded by a polynucleotide chosen from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, 20 SEQ ID NO:l l, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO: 49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO: 57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID 25 NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72 and SEQ ID NO:107. Yet another embodiment of the invention comprises an isolated antibody that binds specifically to an isolated polypeptide of SEQ ID NO: 109. Yet another embodiment of the invention comprises an isolated antibody that binds specifically to an isolated polypeptide of SEQ ID NO: 110. The isolated antibody can be a monoclonal antibody or 30 a polyclonal antibody. 9 WO 01/30972 PCT/USOO/29690 Another embodiment of the invention provides a method for preventing, treating, modulating, or ameliorating a medical condition, such as a neuropsychiatric disorder, comprising administering to a mammalian subject a therapeutically effective amount of a polypeptide of the invention or a polynucleotide of the invention. In one preferred 5 embodiment, a method for preventing, treating, modulating or ameliorating schizophrenia is provided. In another preferred embodiment, a method for preventing, treating, modulating or ameliorating bipolar disorder is provided. In yet another embodiment, a method for preventing, treating, modulating or ameliorating addiction-related behavior is provided. 10 A further embodiment of the invention provides an isolated antibody that binds specifically to the isolated polypeptide of the invention. A preferred embodiment of the invention provides a method for preventing, treating, modulating, or ameliorating a medical condition, such as a neuropsychiatric disorder, comprising administering to a 15 mammalian subject a therapeutically effective amount of the antibody. In one preferred embodiment, a method for preventing, treating, modulating or ameliorating schizophrenia is provided. In another preferred embodiment, a method for preventing, treating, modulating or ameliorating bipolar disorders is provided. In yet another embodiment, a method for preventing, treating, modulating or ameliorating addiction-related behavior is 20 provided. An additional embodiment of the invention provides a method of diagnosing a pathological condition or a susceptibility to a pathological condition in a subject. The method comprises determining the presence or absence of a mutation in a polynucleotide 25 of the invention. A pathological condition or a susceptibility to a pathological condition, such as a neuropsychiatric disorder, is diagnosed based on the presence or absence of the mutation. In one preferred embodiment, a method for diagnosing schizophrenia is provided. In another preferred embodiment, a method for diagnosing bipolar disorders is provided. In yet another embodiment, a method for preventing, treating, modulating or 30 ameliorating addiction-related behavior is provided. 10 WO 01/30972 PCT/USOO/29690 Even another embodiment of the invention provides a method of diagnosing a pathological condition or a susceptibility to a pathological condition, such as a neuropsychiatric disorder, in a subject. Especially preferred embodiments include methods of diagnosing schizophrenia and bipolar disorders. The method comprises 5 detecting an alteration in expression of a polypeptide encoded by the polynucleotide of the invention, wherein the presence of an alteration in expression of the polypeptide is indicative of the pathological condition or susceptibility to the pathological condition. The alteration in expression can be an increase in the amount of expression or a decrease in the amount of expression. In a preferred embodiment a first biological sample is 10 obtained from a patient suspected of having a neuropsychiatric disorder, for example, schizophrenia, bipolar disorder, or addiction-related behavior, and a second sample from a suitable comparable control source is obtained. The amount of at least one polypeptide encoded by a polynucleotide of the invention is determined in the first and second sample. The amount of the polypeptide in the first and second samples is determined. A 15 patient is diagnosed as having a neuropsychiatric disorder if the amount of the polypeptide in the first sample is greater than or less than the amount of the polypeptide in the second sample. Another embodiment of the invention provides a method for identifying a binding 20 partner to a polypeptide of the invention. A polypeptide of the invention is contacted with a binding partner and it is determined whether the binding partner effects an activity of the polypeptide. Yet another embodiment of the invention is a method of identifying an activity of 25 an expressed polypeptide in a biological assay. A polypeptide of the invention is expressed in a cell and isolated. The expressed polypeptide is tested for an activity in a biological assay and the activity of the expressed polypeptide is identified based on the test results. 30 Still another embodiment of the invention provides a substantially pure isolated DNA molecule suitable for use as a probe for genes regulated in neuropsychiatric 11 WO 01/30972 PCT/USOO/29690 disorders, chosen from the group consisting of the DNA molecules shown in I of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO: 11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID 5 NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO: 49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO: 57, SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72 and SEQ ID NO:107. 10 Even another embodiment of the invention provides a kit for detecting the presence of a polypeptide of the invention in a mammalian tissue sample. The kit comprises a first antibody which immunoreacts with a mammalian protein encoded by a gene corresponding to the polynucleotide of the invention or with a polypeptide encoded 15 by the polynucleotide in an amount sufficient for at least one assay and suitable packaging material. The kit can further comprise a second antibody that binds to the first antibody. The second antibody can be labeled with enzymes, radioisotopes, fluorescent compounds, colloidal metals, chemiluminescent compounds, phosphorescent compounds, or bioluminescent compounds. 20 Another embodiment of the invention provides a kit for detecting the presence of genes encoding a protein comprising a polynucleotide of the invention, or fragment thereof having at least 10 contiguous bases, in an amount sufficient for at least one assay, and suitable packaging material. 25 Yet another embodiment of the invention provides a method for detecting the presence of a nucleic acid encoding a protein in a mammalian tissue sample. A polynucleotide of the invention or fragment thereof having at least 10 contiguous bases is hybridized with the nucleic acid of the sample. The presence of the hybridization product 30 is detected. 12 WO 01/30972 PCT/USOO/29690 BRIEF DESCRIPTION OF THE DRAWINGS These and other features, aspects, and advantages of the present invention will become better understood with reference to the following description, appended claims, 5 and accompanying drawings where: Figure 1 is a graphical representation of the results of TOGA analysis using a 5' PCR primer with parsing bases AGTA, showing PCR products produced from mRNA extracted from the striatum/nucleus accumbens of mice treated with 7.5 mg/kg of 10 clozapine for the following durations: control (no clozapine), 45 minutes, 7 hours, 5 days, 12 days, and 14 days, where the vertical index line indicates a PCR product of about 106 b.p. that is present in the control sample and enriched in the clozapine-treated samples; 15 Figure 2A-C is a graphical representation of a more detailed analysis of the 106 bp PCR product indicated in Figure 1. The upper panel (Figure 2A) shows the PCR product generated with the clone specific primer (SEQ ID NO: 28) and the fluoresecnt labeled universal 3' PCR primer (SEQ ID NO: 23). Figure 2B shows the PCR products produced in the original TOGA reaction using a 5' PCR primer, C-G-A-C-G-G-T-A-T-C 20 G-G-A-G-T-A (SEQ ID NO: 94), and the fluoresecnt labeled universal 3' PCR primer (SEQ ID NO: 23). In the bottom panel (Figure 2C), the traces from the top panel and middle panels are overlaid, demonstrating that the PCR product produced using an extended primer based on the cloned sequence is the same length as the original PCR product; 25 Figure 3 is a graphical representation of the results of TOGA analysis using a 5' PCR primer with parsing bases CACC, showing PCR products produced from mRNA extracted from the striatum/nucleus accumbens of mice treated with 7.5 mg/kg of clozapine for the following durations: control (no clozapine), 45 minutes, 7 hours, 5 30 days, 12 days, and 14 days, where the vertical index line indicates a PCR product of 13 WO 01/30972 PCT/USOO/29690 about 201 b.p. that is present in the control sample and increasingly enriched over time in the clozapine-treated samples; Figure 4 shows a Northern Blot analysis of clone CLZ_5 (CACC 201), where an 5 agarose gel containing poly A enriched mRNA from the striatum/nucleus accumbens of mice treated with clozapine as well as size standards was blotted after electrophoresis and probed with radiolabelled CLZ_5. Mice were treated with clozapine (7.5 mg/kg) for the following time durations before mRNA extraction: control (no clozapine), 45 minutes, 7 hours, 5 days, 12 days, and 14 days; 10 Figure 5 shows a Northern Blot analysis of clone CLZ_5 (CACC 201), where an agarose gel containing poly A enriched mRNA from the striatum/nucleus accumbens of mice treated with haloperidol as well as size standards was blotted after electrophoresis and probed with radiolabelled CLZ_5. Mice were treated with haloperidol (4 mg/kg) for 15 the following time durations before mRNA extraction: control (no haloperidol), 45 minutes, 7 hours, 10 days, and 14 days; Figure 6 is a graphical representation comparing the results of the TOGA analysis of clone CLZ_5 shown in Fig. 3 and the Northern Blot analysis of clone CLZ_5 shown in 20 Figure 4; Figure 7A-C is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_5, showing the pattern of CLZ_5 mRNA expression in mouse anterior brain (7A), midbrain (7B), and posterior brain (7C), where CLZ_5 is 25 expressed in scattered glial cells and white matter tracts; Figure 8A-I is an in situ hybridization analyses, using an antisense cRNA probe directed against the 3' end of CLZ_5, showing CLZ_5 mRNA expression in mouse anterior brain (8A-C), midbrain (8D-F), and posterior brain (8G-I) in saline-treated mice 30 (top row), mice treated with clozapine for 5 days (middle row), and mice treated with 14 WO 01/30972 PCT/USOO/29690 clozapine for 14 days (bottom row), where the clozapine treatment induces expression in the glial cells; Figure 9A-H shows a darkfield photomicrograph of various brain regions, 5 including the corpus callosum (cc, Fig. 9A, E); caudate putamen (CPu, Fig. 9B, F); anterior commissure (aca, Fig. 9C, G); and globus pallidus (GP, Fig. 9D, H) in control (9A-D) and clozapine-treated (9E-H) animals; Figure 1 OA-D shows a darkfield photomicrograph in the internal capsule (ic) 10 (10A, B) and a brightfield view of the optic tract (opt) (10C, D) from control (1OA, C) and clozapine-treated (1 OB, D) animals; Figure 11A-H shows GFAP and apoD co-localization in the striatum (11 A, B, D, E) and optic tract (1 C, F) of control saline (1 IA, B, C) and clozapine-treated animals 15 (11D, E, F), with thick arrows designating the co-localization of GFAP and apoD mRNA and thin arrows designating the expression of apoD only; 1IG-H shows apoD immunohistochemistry with an anti-human apoD primary antibody (Novocastra, Newcastle, UK) in the optic tract of control saline (11 G) and clozapine-treated animals (11H). 20 Figure 12 shows a Northern Blot analysis of clone CLZ_5, where an agarose gel containing poly A enriched mRNA from cultured glial cells treated with clozapine as well as size standards was blotted after electrophoresis and probed with radiolabelled CLZ_5. Cultured glial cells were treated with different concentrations of clozapine for 25 different lengths of time before mRNA extraction as follows: A= control (no clozapine), B= 100 nM clozapine, 1 day, C= I1pM clozapine, 1 day, D= 100 nM clozapine, 1 week, E= 1pM clozapine, 1 week; Figure 13 is a graphical representation of the results of TOGA analysis using a 5' 30 PCR primer with parsing bases TTGT, showing PCR products produced from mRNA extracted from the striatum/nucleus accumbens of mice treated with 7.5 mg/kg clozapine 15 WO 01/30972 PCT/USOO/29690 as follows: control (no clozapine), 45 minutes, 7 hours, 5 days, 12 days, and 14 days, where the vertical index line indicates a PCR product of about 266 b.p. that is present in the control sample, is down-regulated within 45 minutes in the clozapine-treated sample, and remains down-regulated for 14 days in the presence of clozapine; 5 Figure 14 is a graphical representation of the results of TOGA analysis using a 5' PCR primer with parsing bases TTGT, showing PCR products produced from mRNA extracted from the brain of morphine-treated mice as follows: control striatum (PS), acutely treated striatum (AS), withdrawal striatum (WS), control amygdala (PA), acutely 10 treated amygdala (AA), chronically treated amygdala (TA), and withdrawal amygdala (WA), where the vertical index line indicates a PCR product of about 266 b.p. that is more abundant in control striatum than control amygdala and is differentially regulated by morphine in striatum versus amygdala; 15 Figure 15 shows a Northern Blot analysis of clone CLZ_40 (TTGT 266), where an agarose gel containing poly A enriched mRNA from the striatum/nucleus accumbens of clozapine-treated mice as well as size standards was blotted after electrophoresis and probed with radiolabelled CLZ_40. Mice were treated with clozapine (7.5 mg/kg) for the following time durations before mRNA extraction: control (no clozapine), 45 minutes, 7 20 hours, 5 days, 12 days, and 14 days; Figure 16 is a graphical representation comparing the results of the TOGA analysis of clone CLZ_40 shown in Fig. 13 and the Northern Blot analysis of clone CLZ_40 shown in Figure 15; 25 Figure 17A-B is an in situ hybridization analysis, showing clone CLZ_40 mRNA expression in mouse brain using an antisense cRNA probe directed against the 3' end of CLZ_40, where 17A shows expression in the nucleus accumbens (Acb) and pyriform cortex (Pir) and 17B shows expression in the dentate gyrus (DG); 30 16 WO 01/30972 PCT/USOO/29690 Figure 18 is a graphical representation of the results of TOGA analysis using a 5' PCR primer with parsing bases TATT, showing PCR products produced from mRNA extracted from the striatum/nucleus accumbens of mice treated with 7.5 mg/kg clozapine as follows: control (no clozapine), 45 minutes, 7 hours, 5 days, 12 days, and 14 days, 5 where the vertical index line indicates a PCR product of about 89 b.p. that is present in the control sample and is differentially regulated by clozapine treatment over time. Figure 19 shows the consensus sequence from the cluster of the following 4 sequences: A1415388: Soares mouse p3NMF19.5 Mus musculus cDNA clone 10 IMAGE:350746 3', mRNA sequence; A1841003: UI-M-AMO-ado-e-04-0-UI.s1 NIHBMAPMAM Mus musculus cDNA clone UI-M-AM0-ado-e-04-0-UI 3', mRNA sequence; A1413353: Soares mouse embryo NbME13.5 14.5 Mus musculus cDNA IMAGE:356159 3', mRNA sequence; A1425991: Soares mouse embryo NbME13.5 14.5 Mus musculus cDNA IMAGE:426077 3', mRNA sequence. 15 Figure 20 shows the sequence of the EST AF006196: Mus musculus metalloprotease-disintegrin MDC 15 mRNA, complete cds. Figure 21 shows the the consensus sequence from the cluster of the following 3 20 sequences: C86593: Mus musculus fertilized egg cDNA 3'-end sequence, clone J0229E09 3', mRNA sequence; A1428410: Life Tech mouse embryo 13 5dpc 10666014 Mus musculus cDNA clone IMAGE:553802 3', mRNA sequence; A1561814: Stratagene mouse skin (#937313) Mus musculus cDNA clone IMAGE:1227449 3', mRNA sequence. 25 Figure 22 is a graphical representation of a Northern Blot analysis of clone CLZ_44 (ACGG 352), where an agarose gel containing poly A enriched mRNA from the striatum/nucleus accumbens of clozapine-treated mice as well as size standards was blotted after electrophoresis and probed with radiolabelled CLZ_44. Mice were treated 30 with clozapine (7.5 mg/kg), haloperidol (4 mg/kg), or ketanserin (4 mg/kg) for two weeks before mRNA extraction. 17 WO 01/30972 PCT/USOO/29690 Figure 23 is a graphical representation of a Northern Blot analysis of clone CLZ_38 (TGCA 109), where an agarose gel containing poly A enriched mRNA from the striatum/nucleus accumbens of clozapine-treated mice as well as size standards was 5 blotted after electrophoresis and probed with radiolabelled CLZ_38. Mice were treated with clozapine (7.5 mg/kg) for the following time durations before mRNA extraction: control (no clozapine), 45 minutes, 7 hours, 5 days, 12 days, and 14 days; Figure 24A-B is an in situ hybridization analysis using an antisense cRNA probe 10 directed against the 3' end of CLZ_ 16, showing the pattern of CLZ_16 mRNA expression in coronal sections through hemispheres in mouse brain. Figure 24A shows dense labelling in the cortex and surrounding the hippocampal formation as well as moderate labelling in the dorsal thalamus and posterior brain. Figure 24B shows uniform labelling throughout; 15 Figure 25A-B is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_17, showing the pattern of CLZ_17 mRNA expression in a coronal section through the hemispheres (25A) and cross section through the midbrain (25B) in mouse brain; 20 Figure 26A-B is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_24, showing the pattern of CLZ_24 mRNA expression in a coronal section through the hemispheres (26A) and cross section through the brainstem (26B) in mouse brain; 25 Figure 27A-B is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_26, showing the pattern of CLZ_26 mRNA expression in a coronal section of the hemispheres at the level of hippocampal formation (27A) and coronal section of the hemispheres at the level of striatum (27B) in mouse 30 brain; 18 WO 01/30972 PCT/USOO/29690 Figure 28A-B is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_28, showing the pattern of CLZ_28 mRNA expression in a coronal section through the hemispheres at the level of hippocampus (28A) and coronal section through the posterior region of hemispheres (28B) in mouse 5 brain; Figure 29A-B is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_3, showing the pattern of CLZ_3 mRNA expression in a coronal section through the hemispheres at level of hippocampus (29A) and cross 10 section through midbrain (29B) in mouse brain; Figure 30A-B is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_34, showing the pattern of CLZ_34 mRNA expression in a coronal section through the hemispheres at the level of hippocampus 15 (30A) and cross section through the midbrain (30B) in mouse brain; Figure 31A-C is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_43, showing the pattern of CLZ_43 mRNA expression in coronal sections of the hemispheres showing labelling in the striatum 20 (31A), labelling in the cortex (31B), and intense labelling in the striatum (31C) in mouse brain; Figure 32A-B is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_44, showing the pattern of CLZ_44 mRNA 25 expression in a coronal section showing labelling in the hippocampus, hypothalamus, and temporal cortex (32A) and coronal section showing cortical labelling (32B) in mouse brain; Figure 33A-B is an in situ hybridization analysis using an antisense cRNA probe 30 directed against the 3' end of CLZ_64, showing the pattern of CLZ_64 mRNA expression in different coronal sections of the hemispheres in mouse brain. 19 WO 01/30972 PCT/USOO/29690 DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS 5 Definitions The following definitions are provided to facilitate understanding of certain terms used throughout this specification. 10 In the present invention, "isolated" refers to material removed from its original environment (e.g., the natural environment if it is naturally occurring), and thus is altered "by the hand of man" from its natural state. For example, an isolated polynucleotide could be part of a vector or a composition of matter, or could be contained within a cell, and still be "isolated" because that vector, composition of matter, or particular cell is not 15 the original environment of the polynucleotide. In the present invention, a "secreted" protein refers to those proteins capable of being directed to the ER, secretory vesicles, or the extracellular space as a result of a signal sequence, as well as those proteins released into the extracellular space without 20 necessarily containing a signal sequence. If the secreted protein is released into the extracellular space, the secreted protein can undergo extracellular processing to produce a "mature" protein. Release into the extracellular space can occur by many mechanisms, including exocytosis and proteolytic cleavage. 25 As used herein, a "polynucleotide" refers to a molecule having a nucleic acid sequence contained in SEQ ID NOs: 1-19; 49-52; 57-72 and 107. For example, the polynucleotide can contain all or part of the nucleotide sequence of the full length cDNA sequence, including the 5' and 3' untranslated sequences, the coding region, with or without the signal sequence, the secreted protein coding region, as well as fragments, 30 epitopes, domains, and variants of the nucleic acid sequence. Moreover, as used herein, a "polypeptide" refers to a molecule having the translated amino acid sequence generated from the polynucleotide as broadly defined. 20 WO 01/30972 PCT/USOO/29690 A "polynucleotide" of the present invention also includes those polynucleotides capable of hybridizing, under stringent hybridization conditions, to sequences contained in SEQ ID NOs: 1-19; 49-52; 57-72 and 107, or the complement thereof, or the cDNA. "Stringent hybridization conditions" refers to an overnight incubation at 420 C in a 5 solution comprising 50% formamide, 5x SSC (750 mM NaCl, 75 mM sodium citrate), 50 mM sodium phosphate (pH 7.6), 5x Denhardt's solution, 10% dextran sulfate, and 20 pg/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0.1x SSC at about 65*C. 10 Also contemplated are nucleic acid molecules that hybridize to the polynucleotides of the present invention at lower stringency hybridization conditions. Changes in the stringency of hybridization and signal detection are primarily accomplished through the manipulation of formamide concentration (lower percentages of formamide result in lowered stringency); salt conditions, or temperature. For example, 15 lower stringency conditions include an overnight incubation at 37*C in a solution comprising 6X SSPE (20X SSPE = 3M NaCl; 0.2M NaH 2

PO

4 ; 0.02M EDTA, pH 7.4), 0.5% SDS, 30% formamide, 100 ug/ml salmon sperm blocking DNA; followed by washes at 50*C with 1XSSPE, 0.1% SDS. In addition, to achieve even lower stringency, washes performed following stringent hybridization can be done at 20 higher salt concentrations (e.g. 5X SSC). Note that variations in the above conditions may be accomplished through the inclusion and/or substitution of alternate blocking reagents used to suppress background in hybridization experiments. Typical blocking reagents include Denhardt's reagent, 25 BLOTTO, heparin, denatured salmon sperm DNA, and commercially available proprietary formulations. The inclusion of specific blocking reagents may require modification of the hybridization conditions described above, due to problems with compatibility. 30 Of course, a polynucleotide which hybridizes only to polyA+ sequences (such as any 3' terminal polyA+ tract of a cDNA shown in the sequence listing), or to a 21 WO 01/30972 PCT/USOO/29690 complementary stretch of T (or U) residues, would not be included in the definition of "polynucleotide," since such a polynucleotide would hybridize to any nucleic acid molecule containing a poly (A) stretch or the complement thereof (e.g., practically any double-stranded cDNA clone). 5 A polynucleotide of the present invention can be composed of any polyribonucleotide or polydeoxribonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA. For example, polynucleotides can be composed of single and double-stranded DNA, DNA that is a mixture of single- and double-stranded regions, 10 single- and double-stranded RNA, and RNA that is mixture of single- and double stranded regions, hybrid molecules comprising DNA and RNA that may be single stranded or, more typically, double-stranded or a mixture of single- and double-stranded regions. In addition, the polynucleotide can be composed of triple-stranded regions comprising RNA or DNA or both RNA and DNA. A polynucleotide may also contain 15 one or more modified bases or DNA or RNA backbones modified for stability or for other reasons. "Modified" bases include, for example, tritylated bases and unusual bases such as inosine. A variety of modifications can be made to DNA and RNA; thus, "polynucleotide" embraces chemically, enzymatically, or metabolically modified forms. 20 The polypeptide of the present invention can be composed of amino acids joined to each other by peptide bonds or modified peptide bonds, i.e., peptide isosteres, and may contain amino acids other than the 20 gene-encoded amino acids. The polypeptides may be modified by either natural processes, such as posttranslational processing, or by chemical modification techniques which are well known in the art. Such modifications 25 are well described in basic texts and in more detailed monographs, as well as in a voluminous research literature. Modifications can occur anywhere in a polypeptide, including the peptide backbone, the amino acid side-chains and the amino or carboxyl termini. It will be appreciated that the same type of modification may be present in the same or varying degrees at several sites in a given polypeptide. Also, a given 30 polypeptide may contain many types of modifications. Polypeptides may be branched, for example, as a result of ubiquitination, and they may be cyclic, with or without 22 WO 01/30972 PCT/USOO/29690 branching. Cyclic, branched, and branched cyclic polypeptides may result from posttranslation natural processes or may be made by synthetic methods. Modifications include acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or 5 nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formulation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, pegylation, proteolytic 10 processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer RNA mediated addition of amino acids to proteins such as arginylation, and ubiquitination. (See, e.g., T. E. Creighton, Proteins - Structure And Molecular Properties, 2nd Ed., W. H. Freeman and Company, New York (1993); B. C. Johnson, Ed., Posttranslational Covalent Modification Of Proteins, Academic Press, New York, 15 pgs. 1-12 (1983); Seifter et al., Meth. Enzymol, 182:626-646 (1990); Rattan et al., Ann. N.Y Acad. Sci. 663:48-62 (1992)). "A polypeptide having biological activity" refers to polypeptides exhibiting activity similar, but not necessarily identical to, an activity of a polypeptide of the present 20 invention, including mature forms, as measured in a particular biological assay, with or without dose dependency. In the case where dose dependency does exist, it need not be identical to that of the polypeptide, but rather substantially similar to the dose dependence in a given activity as compared to the polypeptide of the present invention (i.e., the candidate polypeptide will exhibit greater activity or not more than about 25-fold 25 less and, preferably, not more than about tenfold less activity, and most preferably, not more than about three-fold less activity relative to the polypeptide of the present invention). The translated amino acid sequence, beginning with the methionine, is identified 30 although other reading frames can also be easily translated using known molecular 23 WO 01/30972 PCT/USOO/29690 biology techniques. The polypeptides produced by the translation of these alternative open reading frames are specifically contemplated by the present invention. SEQ ID NOs: 1-19; 49-52; 57-72 and 107 and the translations of SEQ ID NOs: 1 5 19; 49-52; 57-72 and 107 are sufficiently accurate and otherwise suitable for a variety of uses well known in the art and described further below. These probes will also hybridize to nucleic acid molecules in biological samples, thereby enabling a variety of forensic and diagnostic methods of the invention. Similarly, polypeptides identified from the translations of SEQ ID NOs: 1-19; 49-52; 57-72 and 107 may be used to generate 10 antibodies which bind specifically to the secreted proteins encoded by the cDNA clones identified. Nevertheless, DNA sequences generated by sequencing reactions can contain sequencing errors. The errors exist as misidentified nucleotides, or as insertions or 15 deletions of nucleotides in the generated DNA sequence. The erroneously inserted or deleted nucleotides cause frame shifts in the reading frames of the predicted amino acid sequence. In these cases, the predicted amino acid sequence diverges from the actual amino acid sequence, even though the generated DNA sequence may be greater than 99.9% identical to the actual DNA sequence (for example, one base insertion or deletion 20 in an open reading frame of over 1000 bases). The present invention also relates to the genes corresponding to SEQ ID NOs: 1 19; 49-52; 57-72 and 107, and translations of SEQ ID NOs: 1-19; 49-52; 57-72 and 107. The corresponding gene can be isolated in accordance with known methods using the 25 sequence information disclosed herein. Such methods include preparing probes or primers from the disclosed sequence and identifying or amplifying the corresponding gene from appropriate sources of genomic material. Also provided in the present invention are species homologues. Species 30 homologues may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source for the desired homologue. 24 WO 01/30972 PCT/USOO/29690 The polypeptides of the invention can be prepared in any suitable manner. Such polypeptides include isolated naturally occurring polypeptides, recombinantly produced polypeptides, synthetically produced polypeptides, or polypeptides produced by a combination of these methods. Means for preparing such polypeptides are well 5 understood in the art. The polypeptides may be in the form of the secreted protein, including the mature form, or may be a part of a larger protein, such as a fusion protein (see below). It is often advantageous to include an additional amino acid sequence which contains secretory or 10 leader sequences, pro-sequences, sequences which aid in purification, such as multiple histidine residues, or an additional sequence for stability during recombinant production. The polypeptides of the present invention are preferably provided in an isolated form, and preferably are substantially purified. A recombinantly produced version of a 15 polypeptide, including the secreted polypeptide, can be substantially purified by the one step method described in Smith and Johnson, Gene 67:31-40 (1988). Polypeptides of the invention also can be purified from natural or recombinant sources using antibodies of the invention raised against the secreted protein in methods which are well known in the art. 20 Signal Sequences Methods for predicting whether a protein has a signal sequence, as well as the cleavage point for that sequence, are available. For instance, the method of McGeoch uses the information from a short N-terminal charged region and a subsequent uncharged 25 region of the complete (uncleaved) protein (Virus Res., 3:271-286 (1985)). The method of von Heinje uses the information from the residues surrounding the cleavage site, typically residues -13 to +2, where +1 indicates the amino terminus of the secreted protein (Nucleic Acids Res., 14:4683-4690 (1986)). Therefore, from a deduced amino acid sequence, a signal sequence and mature sequence can be identified. 30 In the present case, the deduced amino acid sequence of the secreted polypeptide was analyzed by a computer program called Signal P (Nielsen et al., Protein Engineering, 10:1-6 (1997), which predicts the cellular location of a protein based on the amino acid 25 WO 01/30972 PCT/USOO/29690 sequence. As part of this computational prediction of localization, the methods of McGeoch and von Heinje are incorporated. As one of ordinary skill would appreciate, however, cleavage sites sometimes 5 vary from organism to organism and cannot be predicted with absolute certainty. Accordingly, the present invention provides secreted polypeptides having a sequence corresponding to the translations of SEQ. ID NOs: 1-19 which have an N-terminus beginning within 5 residues (i.e., + or - 5 residues) of the predicted cleavage point. Similarly, it is also recognized that in some cases, cleavage of the signal sequence from a 10 secreted protein is not entirely uniform, resulting in more than one secreted species. These polypeptides, and the polynucleotides encoding such polypeptides, are contemplated by the present invention. Moreover, the signal sequence identified by the above analysis may not 15 necessarily predict the naturally occurring signal sequence. For example, the naturally occurring signal sequence may be further upstream from the predicted signal sequence. However, it is likely that the predicted signal sequence will be capable of directing the secreted protein to the ER. These polypeptides, and the polynucleotides encoding such polypeptides, are contemplated by the present invention. 20 Polynucleotide and Polypeptide Variants "Variant" refers to a polynucleotide or polypeptide differing from the polynucleotide or polypeptide of the present invention, but retaining essential properties thereof. Generally, variants are overall closely similar, and, in many regions, identical to 25 the polynucleotide or polypeptide of the present invention. "Identity" per se has an art-recognized meaning and can be calculated using published techniques. (See, e.g., Lesk, A.M., Ed., Computational Molecular Biology, Oxford University Press, New York, (1988); Smith, D.W., Ed, Biocomputing: 30 Informatics And Genome Projects, Academic Press, New York, (1993); Griffin, A.M., and Griffin, H.G., Eds., Computer Analysis Of Sequence Data, Part I, Humana Press, New Jersey, (1994); von Heinje, G., Sequence Analysis In Molecular Biology, Academic 26 WO 01/30972 PCT/USOO/29690 Press, (1987); and Gribskov, M. and Devereux, J., Eds., Sequence Analysis Primer, M Stockton Press, New York, (1991)). While there exists a number of methods to measure identity between two polynucleotide or polypeptide sequences, the term "identity" is well known to skilled artisans. (See, e.g., Carillo, H., and Lipton, D., SIAM J. Applied Math., 5 48:1073 (1988)). Methods commonly employed to determine identity or similarity between two sequences include, but are not limited to, those disclosed in Martin J. Bishop, ed., "Guide to Huge Computers," Academic Press, San Diego, (1994), and Carillo, H., and Lipton, D., SIAM J. Applied Math, 48:1073 (1988)). Methods for aligning polynucleotides or polypeptides are codified in computer programs, including 10 the GCG program package (Devereux, J., et al., Nuc. Acids Res. 12(1):387 (1984)), BLASTP, BLASTN, FASTA (Atschul, S.F. et al., J. Molec. Biol., 215:403 (1990), Bestfit program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science Drive, Madison, WI 53711 (using the local homology algorithm of Smith and Waterman, Advances in Applied 15 Mathematics 2:482-489 (1981)). When using any of the sequence alignment programs to determine whether a particular sequence is, for instance, 95% identical to a reference sequence, the parameters are set so that the percentage of identity is calculated over the full length of the reference 20 polynucleotide and that gaps in identity of up to 5% of the total number of nucleotides in the reference polynucleotide are allowed. A preferred method for determining the best overall match between a query sequence (a sequence of the present invention) and a subject sequence, also referred to as 25 a global sequence alignment, can be determined using the FASTDB computer program based on the algorithm of Brutlag et al. (Comp. App. Biosci., 6:237-245 (1990)) The term "sequence" includes nucleotide and amino acid sequences. In a sequence alignment the query and subject sequences are either both nucleotide sequences or both amino acid sequences. The result of said global sequence alignment is in percent identity. Preferred 30 parameters used in a FASTDB search of a DNA sequence to calculate percent identity are: Matrix=Unitary, k-tuple=4, Mismatch Penalty=l, Joining Penalty=30, Randomization Group Length=0, and Cutoff Score=l, Gap Penalty=5, Gap Size Penalty 0.05, and Window Size=500 or query sequence length in nucleotide bases, whichever is shorter. 27 WO 01/30972 PCT/USOO/29690 Preferred parameters employed to calculate percent identity and similarity of an amino acid alignment are: Matrix=PAM 150, k-tuple=2, Mismatch Penalty= 1, Joining Penalty=20, Randomization Group Length=O, Cutoff Score=l, Gap Penalty=5, Gap Size Penalty=0.05, and Window Size=500 or query sequence length in amino acid residues, 5 whichever is shorter. As an illustration, a polynucleotide having a nucleotide sequence of at least 95% "identity" to a sequence contained in SEQ ID NOs: 1-19; 49-52; 57-72 and 107 means that the polynucleotide is identical to a sequence contained in SEQ ID NOs: 1-19; 49-52; 10 57-72 and 107 or the cDNA except that the polynucleotide sequence may include up to five point mutations per each 100 nucleotides of the total length (not just within a given 100 nucleotide stretch). In other words, to obtain a polynucleotide having a nucleotide sequence at least 95% identical to SEQ ID NOs: 1-19; 49-52; 57-72 and 107, up to 5% of the nucleotides in the sequence contained in SEQ ID NOs: 1-19; 49-52; 57-72 and 107 or 15 the cDNA can be deleted, inserted, or substituted with other nucleotides. These changes may occur anywhere throughout the polynucleotide. Further embodiments of the present invention include polynucleotides having at least 80% identity, more preferably at least 90% identity, and most preferably at least 20 95%, 96%, 97%, 98% or 99% identity to a sequence contained in SEQ ID NOs: 1-19; 49 52; 57-72 and 107. Of course, due to the degeneracy of the genetic code, one of ordinary skill in the art will immediately recognize that a large number of the polynucleotides having at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity will encode a polypeptide identical to an amino acid sequence contained in the translations of SEQ ID 25 NOs: 1-19; 49-52; 57-72 and 107. Similarly, by a polypeptide having an amino acid sequence having at least, for example, 95% "identity" to a reference polypeptide, is intended that the amino acid sequence of the polypeptide is identical to the reference polypeptide except that the 30 polypeptide sequence may include up to five amino acid alterations per each 100 amino acids of the total length of the reference polypeptide. In other words, to obtain a polypeptide having an amino acid sequence at least 95% identical to a reference amino acid sequence, up to 5% of the amino acid residues in the reference sequence may be 28 WO 01/30972 PCT/USOO/29690 deleted or substituted with another amino acid, or a number of amino acids up to 5% of the total amino acid residues in the reference sequence may be inserted into the reference sequence. These alterations of the reference sequence may occur at the amino or carboxy terminal positions of the reference amino acid sequence or anywhere between those 5 terminal positions, interspersed either individually among residues in the reference sequence or in one or more contiguous groups within the reference sequence. Further embodiments of the present invention include polypeptides having at least 80% identity, more preferably at least 85% identity, more preferably at least 90% 10 identity, and most preferably at least 95%, 96%, 97%, 98% or 99% identity to an amino acid sequence contained in translations of SEQ ID NOs: 1-19; 49-52; 57-72 and 107. Preferably, the above polypeptides should exhibit at least one biological activity of the protein. 15 In a preferred embodiment, polypeptides of the present invention include polypeptides having at least 90% similarity, more preferably at least 95% similarity, and still more preferably at least 96%, 97%, 98%, or 99% similarity to an amino acid sequence contained in translations of SEQ ID NOs: 1-19; 49-52; 57-72 and 107. 20 The variants may contain alterations in the coding regions, non-coding regions, or both. Especially preferred are polynucleotide variants containing alterations which produce silent substitutions, additions, or deletions, but do not alter the properties or activities of the encoded polypeptide. Nucleotide variants produced by silent substitutions due to the degeneracy of the genetic code are preferred. Moreover, variants 25 in which 5-10, 1-5, or 1-2 amino acids are substituted, deleted, or added in any combination are also preferred. Polynucleotide variants can be produced for a variety of reasons. For instance, a polynucleotide variant may be produced to optimize codon expression for a particular host, i.e., codons in the human mRNA may be changed to those preferred by a bacterial host such as E. coli). 30 Naturally occurring variants are called "allelic variants," and refer to one of several alternate forms of a gene occupying a given locus on a chromosome of an organism (Lewin, B., Ed., Genes II, John Wiley & Sons, New York (1985)). These 29 WO 01/30972 PCT/USOO/29690 allelic variants can vary at either the polynucleotide and/or polypeptide level. Alternatively, non-naturally occurring variants may be produced by mutagenesis techniques or by direct synthesis. 5 Using known methods of protein engineering and recombinant DNA technology, variants may be generated to improve or alter the characteristics of the polypeptides of the present invention. For instance, one or more amino acids can be deleted from the N terminus or C-terminus of the secreted protein without substantial loss of biological function. Ron et al., reported variant KGF proteins having heparin binding activity even 10 after deleting 3, 8, or 27 amino-terminal amino acid residues (J. Biol. Chem., 268: 2984 2988 (1993)). Similarly, interferon gamma exhibited up to ten times higher activity after deleting 8-10 amino acid residues from the carboxy terminus of this protein (Dobeli et al., J. Biotechnology, 7:199-216 (1988)). 15 Moreover, ample evidence demonstrates that variants often retain a biological activity similar to that of the naturally occurring protein. For example, Gayle et al., conducted extensive mutational analysis of human cytokine IL-1 a (J. Biol. Chem., 268:22105-22111 (1993)). They used random mutagenesis to generate over 3,500 individual IL- I a mutants that averaged 2.5 amino acid changes per variant over the 20 entire length of the molecule. Multiple mutations were examined at every possible amino acid position. The investigators concluded that "[m]ost of the molecule could be altered with little effect on either [binding or biological activity]." (See Gayle et al., (1993), Abstract.) In fact, only 23 unique amino acid sequences, out of more than 3,500 nucleotide sequences examined, produced a protein that significantly differed in activity 25 from wild-type. Furthermore, even if deleting one or more amino acids from the N-terminus or C terminus of a polypeptide results in modification or loss of one or more biological functions, other biological activities may still be retained. For example, the ability of a 30 deletion variant to induce and/or to bind antibodies which recognize the secreted form will likely be retained when less than the majority of the residues of the secreted form are removed from the N-terminus or C-terminus. Whether a particular polypeptide lacking 30 WO 01/30972 PCT/USOO/29690 N- or C-terminal residues of a protein retains such immunogenic activities can readily be determined by routine methods described herein and otherwise known in the art. Thus, the invention further includes polypeptide variants which show substantial 5 biological activity. Such variants include deletions, insertions, inversions, repeats, and substitutions selected according to general rules known in the art so as have little effect on activity. For example, guidance concerning how to make phenotypically silent amino acid substitutions is provided in Bowie, et al., Science, 247:1306-1310 (1990), wherein the authors indicate that there are two main strategies for studying the tolerance of an 10 amino acid sequence to change. The first strategy exploits the tolerance of amino acid substitutions by natural selection during the process of evolution. By comparing amino acid sequences in different species, conserved amino acids can be identified. These conserved amino acids 15 are likely important for protein function. In contrast, the amino acid positions where substitutions have been tolerated by natural selection indicates that these positions are not critical for protein function. Thus, positions tolerating amino acid substitution could be modified while still maintaining biological activity of the protein. 20 The second strategy uses genetic engineering to introduce amino acid changes at specific positions of a cloned gene to identify regions critical for protein function. For example, site directed mutagenesis or alanine-scanning mutagenesis (the introduction of single alanine mutations at every residue in the molecule) can be used (Cunningham and Wells, Science, 244:1081-1085 (1989)). The resulting mutant molecules can then be 25 tested for biological activity. According to Bowie et al., these two strategies have revealed that proteins are surprisingly tolerant of amino acid substitutions. The authors further indicate which amino acid changes are likely to be permissive at certain amino acid positions in the 30 protein. For example, most buried (within the tertiary structure of the protein) amino acid residues require nonpolar side chains, whereas few features of surface side chains are generally conserved. Moreover, tolerated conservative amino acid substitutions involve replacement of the aliphatic or hydrophobic amino acids Ala, Val, Leu and lie; 31 WO 01/30972 PCT/USOO/29690 replacement of the hydroxyl residues Ser and Thr; replacement of the acidic residues Asp and Glu; replacement of the amide residues Asn and Gln, replacement of the basic residues Lys, Arg, and His; replacement of the aromatic residues Phe, Tyr, and Trp; and replacement of the small-sized amino acids Ala, Ser, Thr, Met, and Gly. 5 Besides conservative amino acid substitution, variants of the present invention include (i) substitutions with one or more of the non-conserved amino acid residues, where the substituted amino acid residues may or may not be one encoded by the genetic code, or (ii) substitution with one or more of amino acid residues having a substituent 10 group, or (iii) fusion of the mature polypeptide with another compound, such as a compound to increase the stability and/or solubility of the polypeptide (for example, polyethylene glycol), or (iv) fusion of the polypeptide with additional amino acids, such as an IgG Fc fusion region peptide, or leader or secretory sequence, or a sequence facilitating purification. Such variant polypeptides are deemed to be within the scope of 15 those skilled in the art from the teachings herein. For example, polypeptide variants containing amino acid substitutions of charged amino acids with other charged or neutral amino acids may produce proteins with improved characteristics, such as decreased aggregation. As known, aggregation of 20 pharmaceutical formulations both reduces activity and increases clearance due to the aggregate's immunogenic activity (see, e.g., Pinckard et al., Clin. Exp. Immunol., 2:331 340 (1967); Robbins et al., Diabetes, 36: 838-845 (1987); Cleland et al., Crit. Rev. Therapeutic Drug Carrier Systems, 10:307-377 (1993)). 25 Polynucleotide and Polypeptide Fragments In the present invention, a "polynucleotide fragment" refers to a short polynucleotide having a nucleic acid sequence contained in that shown in SEQ ID NOs: 1-19; 49-52; 57-72 and 107. The short nucleotide fragments are preferably at least about 15 nt, and more preferably at least about 20 nt, still more preferably at least about 30 nt, 30 and even more preferably, at least about 40 nt in length. A fragment "at least 20 nt in length," for example, is intended to include 20 or more contiguous bases from the cDNA sequence contained in that shown in SEQ ID NOs: 1-19; 49-52; 57-72 and 107. These 32 WO 01/30972 PCT/USOO/29690 nucleotide fragments are useful as diagnostic probes and primers as discussed herein. Of course, larger fragments (e.g., 50, 150, and more nucleotides) are preferred. Moreover, representative examples of polynucleotide fragments of the invention, 5 include, for example, fragments having a sequence from about nucleotide number 1-50, 51-100, 101-150, 151-200,201-250,251-300,301-350,351-400,401-450, to the end of SEQ ID NOs: 1-19; 49-52; 57-72 and 107. In this context "about" includes the particularly recited ranges, larger or smaller by several (5, 4, 3, 2, or 1) nucleotides, at either terminus or at both termini. Preferably, these fragments encode a polypeptide 10 which has biological activity. In the present invention, a "polypeptide fragment" refers to a short amino acid sequence contained in the translations of SEQ ID NOs: 1-19; 49-52; 57-72 and 107. Protein fragments may be "free-standing," or comprised within a larger polypeptide of 15 which the fragment forms a part or region, most preferably as a single continuous region. Representative examples of polypeptide fragments of the invention, include, for example, fragments from about amino acid number 1-20, 21-40, 41-60, or 61 to the end of the coding region. Moreover, polypeptide fragments can be about 20, 30, 40, 50 or 60, amino acids in length. In this context "about" includes the particularly recited ranges, 20 larger or smaller by several (5, 4, 3, 2, or 1) amino acids, at either extreme or at both extremes. Preferred polypeptide fragments include the secreted protein as well as the mature form. Further preferred polypeptide fragments include the secreted protein or the mature 25 form having a continuous series of deleted residues from the amino or the carboxy terminus, or both. For example, any number of amino acids, ranging from 1-60, can be deleted from the amino terminus of either the secreted polypeptide or the mature form. Similarly, any number of amino acids, ranging from 1-30, can be deleted from the carboxy terminus of the secreted protein or mature form. Furthermore, any combination 30 of the above amino and carboxy terminus deletions are preferred. Similarly, polynucleotide fragments encoding these polypeptide fragments are also preferred. 33 WO 01/30972 PCT/USOO/29690 Also preferred are polypeptide and polynucleotide fragments characterized by structural or functional domains, such as fragments that comprise alpha-helix and alpha helix forming regions, beta-sheet and beta-sheet-forming regions, turn and turn-forming regions, coil and coil-forming regions, hydrophilic regions, hydrophobic regions, alpha 5 amphipathic regions, beta amphipathic regions, flexible regions, surface-forming regions, substrate binding region, and high antigenic index regions. Polypeptide fragments of the translations of SEQ ID NOs: 1-19; 49-52; 57-72 and 107 falling within conserved domains are specifically contemplated by the present invention. Moreover, polynucleotide fragments encoding these domains are also contemplated. 10 Other preferred fragments are biologically active fragments. Biologically active fragments are those exhibiting activity similar, but not necessarily identical, to an activity of the polypeptide of the present invention. The biological activity of the fragments may include an improved desired activity, or a decreased undesirable activity. 15 Epitopes & Antibodies In the present invention, "epitopes" refer to polypeptide fragments having antigenic or immunogenic activity in an animal, especially in a human. A preferred embodiment of the present invention relates to a polypeptide fragment comprising an 20 epitope, as well as the polynucleotide encoding this fragment. A region of a protein molecule to which an antibody can bind is defined as an "antigenic epitope." In contrast, an "immunogenic epitope" is defined as a part of a protein that elicits an antibody response (see, e.g., Geysen et al., Proc. NatL. Acad. Sci. USA, 81:3998-4002 (1983)). 25 Fragments which function as epitopes may be produced by any conventional means (see, e.g., Houghten, R. A., Proc. NatL. Acad. Sci. USA, 82:5131-5135 (1985), further described in U.S. Patent No. 4,631,211). In the present invention, antigenic epitopes preferably contain a sequence of at 30 least seven, more preferably at least nine, and most preferably between about 15 to about 30 amino acids. Antigenic epitopes are useful to raise antibodies, including monoclonal antibodies, that specifically bind the epitope. (See, for instance, Wilson et al., Cell, 37:767-778 (1984); Sutcliffe, J. G. et al., Science, 219:660-666 (1983)). 34 WO 01/30972 PCT/USOO/29690 Similarly, immunogenic epitopes can be used to induce antibodies according to methods well known in the art. (See, e.g., Sutcliffe et al., supra; Wilson et al., supra; Chow, M. et al., Proc. Nati. Acad. Sci., USA 82:910-914; and Bittle, F. J. et al., J Gen. 5 Virol., 66:2347-2354 (1985)). A preferred immunogenic epitope includes the secreted protein. The immunogenic epitopes may be presented together with a carrier protein, such as an albumin, to an animal system (such as rabbit or mouse) or, if it is long enough (at least about 25 amino acids), without a carrier. However, immunogenic epitopes comprising as few as 8 to 10 amino acids have been shown to be sufficient to raise 10 antibodies capable of binding to, at the very least, linear epitopes in a denatured polypeptide (e.g., in Western blotting.) As used herein, the term "antibody" (Ab) or "monoclonal antibody" (Mab) is meant to include intact molecules as well as antibody fragments (such as, for example, 15 Fab and F(ab')2 fragments) which are capable of specifically binding to protein. Fab and F(ab')2 fragments lack the Fc fragment of intact antibody, clear more rapidly from the circulation, and may have less non-specific tissue binding than an intact antibody (Wahl et al., J. Nucl. Med., 24:316-325 (1983)). Thus, these fragments are preferred, as well as the products of a FAB or other immunoglobulin expression library. Moreover, antibodies 20 of the present invention include chimeric, single chain, and humanized antibodies. Additional embodiments include chimeric antibodies, e.g., humanized versions of murine monoclonal antibodies. Such humanized antibodies may be prepared by known techniques, and offer the advantage of reduced immunogenicity when the antibodies are 25 administered to humans. In one embodiment, a humanized monoclonal antibody comprises the variable region of a murine antibody (or just the antigen binding site thereof) and a constant region derived from a human antibody. Alternatively, a humanized antibody fragment may comprise the antigen binding site of a murine monoclonal antibody and a variable region fragment (lacking the antigen-binding site) 30 derived from a human antibody. Procedures for the production of chimeric and further engineered monoclonal antibodies include those described in Riechmann et al. (Nature, 332:323, 1988), Liu et al. (PNAS, 84:3439, 1987), Larrick et al. (Bio/Technology, 7:934, 1989), and Winter and Harris (TIPS, 14:139, May, 1993). 35 WO 01/30972 PCT/USOO/29690 One method for producing a human antibody comprises immunizing a non-human animal, such as a transgenic mouse, with a polypeptide translated from a nucleotide sequence chosen from SEQ ID NOs: 1-19; 49-52; 57-72 and 107, whereby antibodies 5 directed against the polypeptide translated from a nucleotide sequence chosen from SEQ ID NOs: 1-19; 49-52; 57-72 and 107 are generated in said animal. Procedures have been developed for generating human antibodies in non-human animals. The antibodies may be partially human, or preferably completely human. Non-human animals (such as transgenic mice) into which genetic material encoding one or more human 10 immunoglobulin chains has been introduced may be employed. Such transgenic mice may be genetically altered in a variety of ways. The genetic manipulation may result in human immunoglobulin polypeptide chains replacing endogenous immunoglobulin chains in at least some (preferably virtually all) antibodies produced by the animal upon immunization. Antibodies produced by immunizing transgenic animals with a 15 polypeptide translated from a nucleotide sequence chosen from SEQ ID NOs: 1-19; 49 52; 57-72 and 107 are provided herein. Mice in which one or more endogenous immunoglobulin genes are inactivated by various means have been prepared. Human immunoglobulin genes have been introduced 20 into the mice to replace the inactivated mouse genes. Antibodies produced in the animals incorporate human immunoglobulin polypeptide chains encoded by the human genetic material introduced into the animal. Examples of techniques for production and use of such transgenic animals are described in U.S. Patent Nos. 5,814,318; 5,569,825; and 5,545,806, which are incorporated by reference herein. 25 Monoclonal antibodies may be produced by conventional procedures, e.g., by immortalizing spleen cells harvested from the transgenic animal after completion of the immunization schedule. The spleen cells may be fused with myeloma cells to produce hybridomas by conventional procedures. 30 36 WO 01/30972 PCT/USOO/29690 A method for producing a hybridoma cell line comprises immunizing such a transgenic animal with an immunogen comprising at least seven contiguous amino acid residues of a polypeptide translated from a nucleotide sequence chosen from SEQ ID NOs: 1-19; 49-52; 57-72 and 107; harvesting spleen cells from the immunized animal; 5 fusing the harvested spleen cells to a myeloma cell line, thereby generating hybridoma cells; and identifying a hybridoma cell line that produces a monoclonal antibody that binds a polypeptide translated from a nucleotide sequence chosen from SEQ ID NOs: 1 19; 49-52; 57-72 and 107. Such hybridoma cell lines, and mosclonal antibodies produced therefrom, are encompassed by the present invention. Monoclonal antibodies secreted by 10 the hybridoma cell line are purified by conventional techniques. Antibodies may be employed in an in vitro procedure, or administered in vivo to inhibit biological activity induced by a polypeptide translated from a nucleotide sequence chosen from SEQ ID NOs: 1-19; 49-52; 57-72 and 107. Disorders caused or exacerbated 15 (directly or indirectly) by the interaction of such polypeptides of the present invention with cell surface receptors thus may be treated. A therapeutic method involves in vivo administration of a blocking antibody to a mammal in an amount effective for reducing a biological activity induced by a polypeptide translated from a nucleotide sequence chosen from SEQ ID NOs: 1-19; 49-52; 57-72 and 107. For example, chronic administration of 20 neuroleptics can cause unwanted side effects. Administration of an antibody derived from the identified polynucleotides might block the signaling that causes these side effects. Alternatively, an antibody derived from the identified polynucleotides might selectively block proteins causing motor side effects. 25 Also provided herein are conjugates comprising a detectable (e.g., diagnostic) or therapeutic agent, attached to an antibody directed against a polypeptide translated from a nucleotide sequence chosen from SEQ ID NOs: 1-19; 49-52; 57-72 and 107. Examples of such agents are well known, and include but are not limited to diagnostic radionuclides, therapeutic radionuclides, and cytotoxic drugs. The conjugates find use in 30 in vitro or in vivo procedures. 37 WO 01/30972 PCT/USOO/29690 Fusion Proteins Any polypeptide of the present invention can be used to generate fusion proteins. For example, the polypeptide of the present invention, when fused to a second protein, can be used as an antigenic tag. Antibodies raised against the polypeptide of the present 5 invention can be used to indirectly detect the second protein by binding to the polypeptide. Moreover, because secreted proteins target cellular locations based on trafficking signals, the polypeptides of the present invention can be used as targeting molecules once fused to other proteins. 10 Examples of domains that can be fused to polypeptides of the present invention include not only heterologous signal sequences, but also other heterologous functional regions. The fusion does not necessarily need to be direct, but may occur through linker sequences. 15 Moreover, fusion proteins may also be engineered to improve characteristics of the polypeptide of the present invention. For instance, a region of additional amino acids, particularly charged amino acids, may be added to the N-terminus of the polypeptide to improve stability and persistence during purification from the host cell or subsequent handling and storage. Also, peptide moieties may be added to the polypeptide to 20 facilitate purification. Such regions may be removed prior to final preparation of the polypeptide. The addition of peptide moieties to facilitate handling of polypeptides are familiar and routine techniques in the art. In addition, polypeptides of the present invention, including fragments, and 25 specifically epitopes, can be combined with parts of the constant domain of immunoglobulins (IgG), resulting in chimeric polypeptides. These fusion proteins facilitate purification and show an increased half-life in vivo. One reported example describes chimeric proteins consisting of the first two domains of the human CD4 polypeptide and various domains of the constant regions of the heavy or light chains of 30 mammalian immunoglobulins (see, EP A 394,827; Traunecker et al., Nature, 331:84-86 (1988)). Fusion proteins having disulfide-linked dimeric structures (due to the IgG) can also be more efficient in binding and neutralizing other molecules, than the monomeric 38 WO 01/30972 PCT/USOO/29690 secreted protein or protein fragment alone (Fountoulakis et al., J. Biochem. 270:3958 3964 (1995)). Similarly, EP-A-0 464 533 (Canadian counterpart 2045869) discloses fusion 5 proteins comprising various portions of constant region of immunoglobulin molecules together with another human protein or part thereof. In many cases, the Fc part in a fusion protein is beneficial in therapy and diagnosis, and thus can result in, for example, improved pharmacokinetic properties (see, e.g., EP-A 0 232 262.) Alternatively, deleting the Fc part after the fusion protein has been expressed, detected, and purified, would be 10 desired. For example, the Fc portion may hinder therapy and diagnosis if the fusion protein is used as an antigen for immunizations. In drug discovery, for example, human proteins, such as hIL-5, have been fused with Fe portions for the purpose of high throughput screening assays to identify antagonists of hIL-5 (see, D. Bennett et al., J Molecular Recognition, 8:52-58 (1995); K. Johanson et al., J. Biol. Chem., 270:9459 15 9471 (1995)). Moreover, the polypeptides of the present invention can be fused to marker sequences, such as a peptide which facilitates purification of the fused polypeptide. In preferred embodiments, the marker amino acid sequence is a hexa-histidine peptide, such 20 as the tag provided in a pQE vector (QIAGEN, Inc., 9259 Eton Avenue, Chatsworth, CA, 91311), among others, many of which are commercially available. As described in Gentz et al, for instance, hexa-histidine provides for convenient purification of the fusion protein (Proc. Natl. Acad. Sci. USA, 86:821-824 (1989)). Another peptide tag useful for purification, the "HA" tag, corresponds to an epitope derived from the influenza 25 hemagglutinin protein (Wilson et al., Cell, 37:767 (1984)). Other fusion proteins may use the ability of the polypeptides of the present invention to target the delivery of a biologically active peptide. This might include focused delivery of a toxin to tumor cells, or a growth factor to stem cells. 30 Thus, any of these above fusions can be engineered using the polynucleotides or the polypeptides of the present invention. 39 WO 01/30972 PCT/USOO/29690 Vectors, Host Cells, and Protein Production The present invention also relates to vectors containing the polynucleotide of the present invention, host cells, and the production of polypeptides by recombinant techniques. The vector may be, for example, a phage, plasmid, viral, or retroviral vector. 5 Retroviral vectors may be replication competent or replication defective. In the latter case, viral propagation generally will occur only in complementing host cells. The polynucleotides may be joined to a vector containing a selectable marker for propagation in a host. Generally, a plasmid vector is introduced in a precipitate, such 10 as a calcium phosphate precipitate, or in a complex with a charged lipid. If the vector is a virus, it may be packaged in vitro using an appropriate packaging cell line and then transduced into host cells. The polynucleotide insert should be operatively linked to an appropriate 15 promoter, such as the phage lambda PL promoter, the E. coli lac, trp, phoA and tac promoters, the SV40 early and late promoters and promoters of retroviral LTRs, to name a few. Other suitable promoters will be known to the skilled artisan. The expression constructs will further contain sites for transcription initiation, termination, and, in the transcribed region, a ribosome binding site for translation. The coding portion of the 20 transcripts expressed by the constructs will preferably include a translation initiating codon at the beginning and a termination codon (UAA, UGA or UAG) appropriately positioned at the end of the polypeptide to be translated. As indicated, the expression vectors will preferably include at least one selectable 25 marker. Such markers include dihydrofolate reductase, G418 or neomycin resistance for eukaryotic cell culture and tetracycline, kanamycin or ampicillin resistance genes for culturing in E. coli and other bacteria. Representative examples of appropriate hosts include, but are not limited to, bacterial cells, such as E. coli, Streptomyces and Salmonella typhimurium cells; fungal cells, such as yeast cells; insect cells such as 30 Drosophila S2 and Spodoptera Sf9 cells; animal cells such as CHO, COS, 293, Bowes melanoma cells and plant cells. Appropriate culture mediums and conditions for the above-described host cells are known in the art. 40 WO 01/30972 PCT/USOO/29690 Among vectors preferred for use in bacteria include pQE70, pQE60 and pQE-9, available from QIAGEN, Inc.; pBluescript vectors, Phagescript vectors, pNH8A, PNH16a, pNH18A, pNH46A, available from Stratagene Cloning Systems, Inc.; and ptrc99a, pKK223-3, pKK233-3, pDR540, pRIT5 available from Pharmacia Biotech, Inc. 5 Among preferred eukaryotic vectors are pWLNEO, pSV2CAT, pOG44, pXT1 and pSG available from Stratagene; and pSVK3, pBPV, pMSG and pSVL available from Pharmacia. Other suitable vectors will be readily apparent to the skilled artisan. Introduction of the construct into the host cell can be effected by calcium 10 phosphate transfection, DEAE-dextran mediated transfection, cationic lipid-mediated transfection, electroporation, transduction, infection, or other methods. Such methods are described in many standard laboratory manuals, such as Davis et al., Basic Methods In Molecular Biology, (1986). It is specifically contemplated that the polypeptides of the present invention may in fact be expressed by a host cell lacking a recombinant vector. 15 Currently no specific diagnostic markers exist that can be used to prevent or delay psychotic episodes of schizophrenia. The polynucleotides of the present invention can be used as chromosome markers for diagnosis for schizophrenia. A polypeptide of this invention can be recovered and purified from recombinant cell cultures by well-known 20 methods including ammonium sulfate or ethanol precipitation, acid extraction, anion or cation exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin chromatography. Most preferably, high performance liquid chromatography ("HPLC") is employed for purification. 25 Polypeptides of the present invention, and preferably the secreted form, can also be recovered from: products purified from natural sources, including bodily fluids, tissues and cells, whether directly isolated or cultured; products of chemical synthetic procedures; and products produced by recombinant techniques from a prokaryotic or 30 eukaryotic host, including, for example, bacterial, yeast, higher plant, insect, and mammalian cells. Depending upon the host employed in a recombinant production procedure, the polypeptides of the present invention may be glycosylated or may be non glycosylated. In addition, polypeptides of the invention may also include an initial 41 WO 01/30972 PCT/USOO/29690 modified methionine residue, in some cases as a result of host-mediated processes. Thus, it is well known in the art that the N-terminal methionine encoded by the translation initiation codon generally is removed with high efficiency from any protein after translation in all eukaryotic cells. While the N-terminal methionine on most proteins also 5 is efficiently removed in most prokaryotes, for some proteins, this prokaryotic removal process is inefficient, depending on the nature of the amino acid to which the N-terminal methionine is covalently linked. Uses of the Polynucleotides 10 Each of the polynucleotides identified herein can be used in numerous ways as reagents. The following description should be considered exemplary and utilizes known techniques. The polynucleotides of the present invention are useful for chromosome 15 identification. There exists an ongoing need to identify new chromosome markers, since few chromosome marking reagents, based on actual sequence data (repeat polymorphisms), are presently available. Each polynucleotide of the present invention can be used as a chromosome marker. 20 Briefly, sequences can be mapped to chromosomes by preparing PCR primers (preferably 15-25 bp) from the sequences shown in SEQ ID NOs: 1-19; 49-52; 57-72 and 107. Primers can be selected using computer analysis so that primers do not span more than one predicted exon in the genomic DNA. These primers are then used for PCR screening of somatic cell hybrids containing individual human chromosomes. Only those 25 hybrids containing the human gene corresponding to the SEQ ID NOs: 1-19; 49-52; 57 72 and 107 will yield an amplified fragment. Similarly, somatic hybrids provide a rapid method of PCR mapping the polynucleotides to particular chromosomes. Three or more clones can be assigned per 30 day using a single thermal cycler. Moreover, sublocalization of the polynucleotides can be achieved with panels of specific chromosome fragments. Other gene mapping strategies that can be used include in situ hybridization, prescreening with labeled flow 42 WO 01/30972 PCT/USOO/29690 sorted chromosomes, and preselection by hybridization to construct chromosome specific-cDNA libraries. Precise chromosomal location of the polynucleotides can also be achieved using 5 fluorescence in situ hybridization (FISH) of a metaphase chromosomal spread. This technique uses polynucleotides as short as 500 or 600 bases; however, polynucleotides 2,000-4,000 bp are preferred. For a review of this technique, see Verma et al., Human Chromosomes: a Manual ofBasic Techniques, Pergamon Press, New York (1988). 10 For chromosome mapping, the polynucleotides can be used individually (to mark a single chromosome or a single site on that chromosome) or in panels (for marking multiple sites and/or multiple chromosomes). Preferred polynucleotides correspond to the noncoding regions of the cDNAs because the coding sequences are more likely conserved within gene families, thus increasing the chance of cross hybridization during 15 chromosomal mapping. Once a polynucleotide has been mapped to a precise chromosomal location, the physical position of the polynucleotide can be used in linkage analysis. Linkage analysis establishes coinheritance between a chromosomal location and presentation of a 20 particular disease . Disease mapping data are found, for example in V. McKusick, Mendelian Inheritance in Man (available on line through Johns Hopkins University Welch Medical Library) Assuming one megabase mapping resolution and one gene per 20 kb, a cDNA precisely localized to a chromosomal region associated with the disease could be one of 50-500 potential causative genes. 25 Thus, once coinheritance is established, differences in the polynucleotide and the corresponding gene between affected and unaffected individuals can be examined. The polynucleotides of SEQ ID NOs: 1-19; 49-52; 57-72 and 107 can be used for this analysis of individual humans. 30 First, visible structural alterations in the chromosomes, such as deletions or translocations, are examined in chromosome spreads or by PCR. If no structural alterations exist, the presence of point mutations are ascertained. Mutations observed in 43 WO 01/30972 PCT/USOO/29690 some or all affected individuals, but not in normal individuals, indicates that the mutation may cause the disease. However, complete sequencing of the polypeptide and the corresponding gene from several normal individuals is required to distinguish the mutation from a polymorphism. If a new polymorphism is identified, this polymorphic 5 polypeptide can be used for further linkage analysis. Furthermore, increased or decreased expression of the gene in affected individuals as compared to unaffected individuals can be assessed using polynucleotides of the present invention. Any of these alterations (altered expression, chromosomal 10 rearrangement, or mutation) can be used as a diagnostic or prognostic marker. In addition to the foregoing, a polynucleotide can be used to control gene expression through triple helix formation or antisense DNA or RNA. Both methods rely on binding of the polynucleotide to DNA or RNA. For these techniques, preferred 15 polynucleotides are usually 20 to 40 bases in length and complementary to either the region of the gene involved in transcription (see, Lee et al., NucL. Acids Res., 6:3073 (1979); Cooney et al., Science, 241:456 (1988); and Dervan et al., Science, 251:1360 (1991) for discussion of triple helix formation) or to the mRNA itself (see, Okano, J. Neurochem., 56:560 (1991) and Oligodeoxy-nucleotides as Antisense Inhibitors of Gene 20 Expression, CRC Press, Boca Raton, FL (1988) for a discussion of antisense technique.) Triple helix formation optimally results in a shut-off of RNA transcription from DNA, while antisense RNA hybridization blocks translation of an mRNA molecule into polypeptide. Both techniques are effective in model systems, and the information disclosed herein can be used to design antisense or triple helix polynucleotides in an 25 effort to treat disease. Polynucleotides of the present invention are also useful in gene therapy. One goal of gene therapy is to insert a normal gene into an organism having a defective gene, in an effort to correct the genetic defect. The polynucleotides disclosed in the present 30 invention offer a means of targeting such genetic defects in a highly accurate manner. Another goal is to insert a new gene that was not present in the host genome, thereby producing a new trait in the host cell. 44 WO 01/30972 PCT/USOO/29690 The polynucleotides are also useful for identifying individuals from minute biological samples. The United States military, for example, is considering the use of restriction fragment length polymorphism (RFLP) for identification of its personnel. In this technique, an individual's genomic DNA is digested with one or more restriction 5 enzymes, and probed on a Southern blot to yield unique bands for identifying personnel. This method does not suffer from the current limitations of "Dog Tags" which can be lost, switched, or stolen, making positive identification difficult. The polynucleotides of the present invention can be used as additional DNA markers for RFLP. 10 The polynucleotides of the present invention can also be used as an alternative to RFLP, by determining the actual base-by-base DNA sequence of selected portions of an individual's genome. These sequences can be used to prepare PCR primers for amplifying and isolating such selected DNA, which can then be sequenced. Using this technique, individuals can be identified because each individual will have a unique set of 15 DNA sequences. Once an unique ID database is established for an individual, positive identification of that individual, living or dead, can be made from extremely small tissue samples. Forensic biology also benefits from using DNA-based identification techniques as 20 disclosed herein. DNA sequences taken from very small biological samples such as tissues, e.g., hair or skin, or body fluids, e.g., blood, saliva, semen, etc., can be amplified using PCR. In one prior art technique, gene sequences amplified from polymorphic loci, such as DQa class II HLA gene, are used in forensic biology to identify individuals (Erlich, H., PCR Technology, Freeman and Co. (1992)). Once these specific 25 polymorphic loci are amplified, they are digested with one or more restriction enzymes, yielding an identifying set of bands on a Southern blot probed with DNA corresponding to the DQa class H HLA gene. Similarly, polynucleotides of the present invention can be used as polymorphic markers for forensic purposes. 30 There is also a need for reagents capable of identifying the source of a particular tissue. Such need arises, for example, in forensics when presented with tissue of unknown origin. Appropriate reagents can comprise, for example, DNA probes or primers specific to particular tissue prepared from the sequences of the present invention. 45 WO 01/30972 PCT/USOO/29690 Panels of such reagents can identify tissue by species and/or by organ type. In a similar fashion, these reagents can be used to screen tissue cultures for contamination. In the very least, the polynucleotides of the present invention can be used as 5 molecular weight markers on Southern gels, as diagnostic probes for the presence of a specific mRNA in a particular cell type, as a probe to "subtract-out" known sequences in the process of discovering novel polynucleotides, for selecting and making oligomers for attachment to a "gene chip" or other support, to raise anti-DNA antibodies using DNA immunization techniques, and as an antigen to elicit an immune response. 10 Uses of the Polypeptides Each of the polypeptides identified herein can be used in numerous ways. The following description should be considered exemplary and utilizes known techniques. 15 A polypeptide of the present invention can be used to assay protein levels in a biological sample using antibody-based techniques. For example, protein expression in tissues can be studied with classical immunohistological methods (Jalkanen, M., et al., J. Cell. Biol., 101:976-985 (1985); Jalkanen, M., et al., J. Cell. Biol., 105:3087-3096 (1987)). Other antibody-based methods useful for detecting protein gene expression 20 include immunoassays, such as the enzyme linked immunosorbent assay (ELISA) and the radioimmunoassay (RIA). Suitable antibody assay labels are known in the art and include enzyme labels, such as, glucose oxidase, and radioisotopes, such as iodine (121I, 1211), carbon (1 4 C), sulfur (35S), tritium ( 3 H), indium (11 2 In), and technetium ( 99 mTc), and fluorescent labels, such as fluorescein and rhodamine, and biotin. 25 In addition to assaying secreted protein levels in a biological sample, proteins can also be detected in vivo by imaging. Antibody labels or markers for in vivo imaging of protein include those detectable by X-radiography, NMR or ESR. For X radiography, suitable labels include radioisotopes such as barium or cesium, which emit 30 detectable radiation but are not overtly harmful to the subject. Suitable markers for NMR and ESR include those with a detectable characteristic spin, such as deuterium, which may be incorporated into the antibody by labeling of nutrients for the relevant hybridoma. 46 WO 01/30972 PCT/USOO/29690 A protein-specific antibody or antibody fragment which has been labeled with an appropriate detectable imaging moiety, such as a radioisotope (for example, 1311, m2n, 99 mTc), a radio-opaque substance, or a material detectable by nuclear magnetic resonance, is introduced (for example, parenterally, subcutaneously, or intraperitoneally) into the 5 mammal. It will be understood in the art that the size of the subject and the imaging system used will determine the quantity of imaging moiety needed to produce diagnostic images. In the case of a radioisotope moiety, for a human subject, the quantity of radioactivity injected will normally range from about 5 to 20 millicuries of 99 mTc. The labeled antibody or antibody fragment will then preferentially accumulate at the location 10 of cells which contain the specific protein. In vivo tumor imaging is described in S.W. Burchiel et al., "Immunopharmacokinetics of Radiolabeled Antibodies and Their Fragments" (Chapter 13 in Tumor Imaging: The Radiochemical Detection of Cancer, S.W. Burchiel and B. A. Rhodes, Eds., Masson Publishing Inc. (1982)). 15 Thus, the invention provides a diagnostic method of a disorder, which involves (a) assaying the expression of a polypeptide of the present invention in cells or body fluid of an individual; (b) comparing the level of gene expression with a standard gene expression level, whereby an increase or decrease in the assayed polypeptide gene expression level compared to the standard expression level is indicative of a disorder. Psychiatric 20 disorders and treatment of psychiatric disorders with neuroleptics, including schizophrenia, are associated with a dysregulation of neurotransmitter and/or neuropeptide levels that can result in the up- or down regulation of polynucleotides and polypeptides. These changes can be diagnosed or monitored by assaying changes in polypeptide levels in tissue or fluids such as CSF, blook, or in fecal samples. 25 Moreover, polypeptides of the present invention can be used to treat disease. For example, patients can be administered a polypeptide of the present invention in an effort to replace absent or decreased levels of the polypeptide (e.g., insulin), to supplement absent or decreased levels of a different polypeptide (e.g., hemoglobin S for 30 hemoglobin B), to inhibit the activity of a polypeptide (e.g., an oncogene), to activate the activity of a polypeptide (e.g., by binding to a receptor), to reduce the activity of a membrane bound receptor by competing with it for free ligand (e.g., soluble TNF 47 WO 01/30972 PCT/USOO/29690 receptors used in reducing inflammation), or to bring about a desired response (e.g., blood vessel growth). Similarly, antibodies directed to a polypeptide of the present invention can also be 5 used to treat disease. For example, administration of an antibody directed to a polypeptide of the present invention can bind and reduce overproduction of the polypeptide. Similarly, administration of an antibody can activate the polypeptide, such as by binding to a polypeptide bound to a membrane (receptor). Polypeptides can also be used as antigens to trigger immune responses. 10 Local production of neurotransmitters and neuropeptides modulates many aspects of neuronal function. For example, in schizophrenia overactive neurotransmitter activity is thought to be basis for the psychotic behavior. Administration of an antibody to an overproduced polypeptide can be used to modulate neuronal responses in psychiatric 15 disorders such as schizophrenia. At the very least, the polypeptides of the present invention can be used as molecular weight markers on SDS-PAGE gels or on molecular sieve gel filtration columns using methods well known to those of skill in the art. Polypeptides can also be 20 used to raise antibodies, which in turn are used to measure protein expression from a recombinant cell, as a way of assessing transformation of the host cell. Moreover, the polypeptides of the present invention can be used to test the following biological activities. 25 Biological Activities The polynucleotides and polypeptides of the present invention can be used in assays to test for one or more biological activities. If these polynucleotides and polypeptides do exhibit activity in a particular assay, it is likely that these molecules may be involved in the diseases associated with the biological activity. Thus, the 30 polynucleotides and polypeptides could be used to treat the associated disease. Nervous System Activity 48 WO 01/30972 PCT/USOO/29690 A polypeptide or polynucleotide of the present invention may be useful in treating deficiencies or disorders of the central nervous system or peripheral nervous system, by activating or inhibiting the proliferation, differentiation, or mobilization (chemotaxis) of neuroblasts, stem cells or glial cells. A polypeptide or polynucleotide of the present 5 invention may be useful in treating deficiencies or disorders of the central nervous system or peripheral nervous system, by activating or inhibiting the mechanisms of synaptic transmission, synthesis, metabolism and inactivation of neural transmitters, neuromodulators and trophic factors, expression and incorporation of enzymes, structural proteins, membrane channels and receptors in neurons and glial cells, or altering neural 10 membrane compositions. The etiology of these deficiencies or disorders may be genetic, somatic (such as cancer or some autoimmune disorders), acquired (e.g., by chemotherapy or toxins), or infectious. Moreover, a polynucleotide or polypeptide of the present invention can be 15 used as a marker or detector of a particular nervous system disease or disorder. The disorder or disease can be any of Alzheimer's disease, Pick's disease, Binswanger's disease, other senile dementia, Parkinson's disease, parkinsonism, obsessive compulsive disorders, epilepsy, encephalopathy, ischemia, alcohol addiction, drug addiction, schizophrenia, amyotrophic lateral sclerosis, multiple sclerosis, depression, and bipolar 20 manic-depressive disorder. Alternatively, the polypeptide or polynucleotide of the present invention can be used to study circadian variation, aging, or long-term potentiation, the latter affecting the hippocampus. Additionally, particularly with reference to mRNA species occurring in particular structures within the central nervous system, the polypeptide or polynucleotide of the present invention can be used to study 25 brain regions that are known to be involved in complex behaviors, such as learning and memory, emotion, drug addiction, glutamate neurotoxicity, feeding behavior, olfaction, viral infection, vision, and movement disorders. Immune Activity 30 A polypeptide or polynucleotide of the present invention may be useful in treating deficiencies or disorders of the immune system, by activating or inhibiting the 49 WO 01/30972 PCT/USOO/29690 proliferation, differentiation, or mobilization (chemotaxis) of immune cells. Immune cells develop through a process called hematopoiesis, producing myeloid (platelets, red blood cells, neutrophils, and macrophages) and lymphoid (B and T lymphocytes) cells from pluripotent stem cells. The etiology of these immune deficiencies or disorders may be 5 genetic, somatic, such as cancer or some autoimmune disorders, acquired (e.g., by chemotherapy or toxins), or infectious. Moreover, a polynucleotide or polypeptide of the present invention can be used as a marker or detector of a particular immune system disease or disorder. 10 A polynucleotide or polypeptide of the present invention may be useful in treating or detecting deficiencies or disorders of hematopoietic cells. A polypeptide or polynucleotide of the present invention could be used to increase differentiation and proliferation of hematopoietic cells, including the pluripotent stem cells, in an effort to treat those disorders associated with a decrease in certain (or many) types hematopoietic 15 cells. Examples of immunologic deficiency syndromes include, but are not limited to: blood protein disorders (e.g. aganmaglobulinemia, dysgammaglobulinemia), ataxia telangiectasia, common variable immunodeficiency, Di George's Syndrome, HIV infection, HTLV-BLV infection, leukocyte adhesion deficiency syndrome, lymphopenia, phagocyte bactericidal dysfunction, severe combined immunodeficiency (SCIDs), 20 Wiskott-Aldrich Disorder, anemia, thrombocytopenia, or hemoglobinuria. Moreover, a polypeptide or polynucleotide of the present invention could also be used to modulate hemostatic (the stopping of bleeding) or thrombolytic activity (clot formation). For example, by increasing hemostatic or thrombolytic activity, a 25 polynucleotide or polypeptide of the present invention could be used to treat blood coagulation disorders (e.g., afibrinogenemia, factor deficiencies), blood platelet disorders (e.g. thrombocytopenia), or wounds resulting from trauma, surgery, or other causes. Alternatively, a polynucleotide or polypeptide of the present invention that can decrease hemostatic or thrombolytic activity could be used to inhibit or dissolve clotting. These 30 molecules could be important in the treatment of heart attacks (infarction), strokes, or scarrmng. 50 WO 01/30972 PCT/USOO/29690 A polynucleotide or polypeptide of the present invention may also be useful in treating or detecting autoimmune disorders. Many autoimmune disorders result from inappropriate recognition of self as foreign material by immune cells. This inappropriate recognition results in an immune response leading to the destruction of the host tissue. 5 Therefore, the administration of a polypeptide or polynucleotide of the present invention that inhibits an immune response, particularly the proliferation, differentiation, or chemotaxis of T-cells or in some ways results in the induction of tolerance, may be an effective therapy in preventing autoimmune disorders. 10 Examples of autoimmune disorders that can be treated or detected by the present invention include, but are not limited to: Addison's Disease, hemolytic anemia, antiphospholipid syndrome, rheumatoid arthritis, dermatitis, allergic encephalomyelitis, glomerulonephritis, Goodpasture's Syndrome, Graves' Disease, Multiple Sclerosis, Myasthenia Gravis, Neuritis, Ophthalmia, Bullous Pemphigoid, Pemphigus, 15 Polyendocrinopathies, Purpura, Reiter's Disease, Stiff-Man Syndrome, Autoimmune Thyroiditis, Systemic Lupus Erythematosus, Autoimmune Pulmonary Inflammation, Guillain-Barre Syndrome, insulin dependent diabetes mellitis, and autoimmune inflammatory eye disease. Schizophrenia has several aspects that suggest an autoimmune component to the disease process. Patients with schizophrenia exhibit immunological 20 abnormalities including hypersecretion of cytokines, presence of antinuclear, anticytoplasmic and antiphospholipid antibodies and a decreased ratio of CD4+/CD8+ cells. Similarly, allergic reactions and conditions, such as asthma (particularly allergic 25 asthma) or other respiratory problems, may also be treated by a polypeptide or polynucleotide of the present invention. Moreover, these molecules can be used to treat anaphylaxis, hypersensitivity to an antigenic molecule, or blood group incompatibility. A polynucleotide or polypeptide of the present invention may also be used to treat 30 and/or prevent organ rejection or graft-versus-host disease (GVHD). Organ rejection occurs by host immune cell destruction of the transplanted tissue through an immune response. Similarly, an immune response is also involved in GVHD, but, in this case, the foreign transplanted immune cells destroy the host tissues. The administration of a 51 WO 01/30972 PCT/USOO/29690 polypeptide or polynucleotide of the present invention that inhibits an immune response, particularly the proliferation, differentiation, or chemotaxis of T-cells, may be an effective therapy in preventing organ rejection or GVHD. 5 Similarly, a polypeptide or polynucleotide of the present invention may also be used to modulate inflammation. For example, the polypeptide or polynucleotide may inhibit the proliferation and differentiation of cells involved in an inflammatory response. These molecules can be used to treat inflammatory conditions, both chronic and acute conditions, including inflammation associated with infection (e.g., septic shock, sepsis, or 10 systemic inflammatory response syndrome (SIRS)), ischemia-reperfusion injury, endotoxin lethality, arthritis, complement-mediated hyperacute rejection, nephritis, cytokine or chemokine induced lung injury, inflammatory bowel disease, Crohn's disease, or resulting from over production of cytokines (e.g., TNF or IL-1). 15 Hyperproliferative Disorders A polypeptide or polynucleotide can be used to treat or detect hyperproliferative disorders, including neoplasms. A polypeptide or polynucleotide of the present invention may inhibit the proliferation of the disorder through direct or indirect interactions. Alternatively, a polypeptide or polynucleotide of the present invention may proliferate 20 other cells which can inhibit the hyperproliferative disorder. For example, by increasing an immune response, particularly increasing antigenic qualities of the hyperproliferative disorder or by proliferating, differentiating, or mobilizing T-cells, hyperproliferative disorders can be treated. This immune response 25 may be increased by either enhancing an existing immune response, or by initiating a new immune response. Alternatively, decreasing an immune response may also be a method of treating hyperproliferative disorders, such as a chemotherapeutic agent. Examples of hyperproliferative disorders that can be treated or detected by a 30 polynucleotide or polypeptide of the present invention include, but are not limited to neoplasms located in the: abdomen, bone, breast, digestive system, liver, pancreas, peritoneum, endocrine glands (adrenal, parathyroid, pituitary, testicles, ovary, thymus, 52 WO 01/30972 PCT/USOO/29690 thyroid), eye, head and neck, nervous (central and peripheral), lymphatic system, pelvic region, skin, soft tissue, spleen, thoracic region, and urogenital system. Similarly, other hyperproliferative disorders can also be treated or detected by a 5 polynucleotide or polypeptide of the present invention. Examples of such hyperproliferative disorders include, but are not limited to: hypergammaglobulinemia, lymphoproliferative disorders, paraproteinemias, purpura, sarcoidosis, Sezary Syndrome, Waldenstron's Macroglobulinemia, Gaucher's Disease, histiocytosis, and any other hyperproliferative disease, besides neoplasia, located in an organ system listed above. 10 Infectious Disease A polypeptide or polynucleotide of the present invention can be used to treat or detect infectious agents. For example, by increasing the immune response, particularly increasing the proliferation and differentiation of B and/or T cells, infectious diseases 15 may be treated. The immune response may be increased by either enhancing an existing immune response, or by initiating a new immune response. Alternatively, the polypeptide or polynucleotide of the present invention may also directly inhibit the infectious agent, without necessarily eliciting an immune response. In the case of schizophrenia, where infectious agents may contribute to the pathology, treatment of 20 patients with a polypeptide or polynucleotide of the present invention might act as a vaccine to trigger a more efficient immune response, altering the course of disease. Viruses are one example of an infectious agent that can cause disease or symptoms that can be treated or detected by a polynucleotide or polypeptide of the 25 present invention. Examples of viruses, include, but are not limited to the following DNA and RNA viral families: Arbovirus, Adenoviridae, Arenaviridae, Arterivirus, Birnaviridae, Bunyaviridae, Caliciviridae, Circoviridae, Coronaviridae, Flaviviridae, Hepadnaviridae (Hepatitis), Herpesviridae (such as, Cytomegalovirus, Herpes Simplex, Herpes Zoster), Mononegavirus (e.g., Paramyxoviridae, Morbillivirus, Rhabdoviridae), 30 Orthomyxoviridae (e.g., Influenza), Papovaviridae, Parvoviridae, Picornaviridae, Poxviridae (such as Smallpox or Vaccinia), Reoviridae (e.g., Rotavirus), Retroviridae (HTLV-I, HTLV-II, Lentivirus), and Togaviridae (e.g., Rubivirus). Viruses falling within these families can cause a variety of diseases or symptoms, including, but not 53 WO 01/30972 PCT/USOO/29690 limited to: arthritis, bronchiollitis, encephalitis, eye infections (e.g., conjunctivitis, keratitis), chronic fatigue syndrome, hepatitis (A, B, C, E, Chronic Active, Delta), meningitis, opportunistic infections (e.g., AIDS), pneumonia, Burkitt's Lymphoma, chickenpox, hemorrhagic fever, Measles, Mumps, Parainfluenza, Rabies, the common 5 cold, Polio, leukemia, Rubella, sexually transmitted diseases, skin diseases (e.g., Kaposi's, warts), and viremia. A polypeptide or polynucleotide of the present invention can be used to treat or detect any of these symptoms or diseases. Similarly, bacterial or fungal agents that can cause disease or symptoms and that 10 can be treated or detected by a polynucleotide or polypeptide of the present invention include, but not limited to, the following Gram-Negative and Gram-positive bacterial families and fungi: Actinomycetales (e.g., Corynebacterium, Mycobacterium, Norcardia), Aspergillosis, Bacillaceae (e.g., Anthrax, Clostridium), Bacteroidaceae, Blastomycosis, Bordetella, Borrelia, Brucellosis, Candidiasis, Campylobacter, Coccidioidomycosis, 15 Cryptococcosis, Dermatocycoses, Enterobacteriaceae (Klebsielia, Salmonella, Serratia, Yersinia), Erysipelothrix, Helicobacter, Legionellosis, Leptospirosis, Listeria, Mycoplasmatales, Neisseriaceae (e.g., Acinetobacter, Gonorrhea, Menigococcal), Pasteurellacea Infections (e.g., Actinobacillus, Heamophilus, Pasteurella), Pseudomonas, Rickettsiaceae, Chlamydiaceae, Syphilis, and Staphylococcal. These bacterial or fungal 20 families can cause the following diseases or symptoms, including, but not limited to: bacteremia, endocarditis, eye infections (conjunctivitis, tuberculosis, uveitis), gingivitis, opportunistic infections (e.g., AIDS related infections), paronychia, prosthesis-related infections, Reiter's Disease, respiratory tract infections, such as Whooping Cough or Empyema, sepsis, Lyme Disease, Cat-Scratch Disease, Dysentery, Paratyphoid Fever, 25 food poisoning, Typhoid, pneumonia, Gonorrhea, meningitis, Chlamydia, Syphilis, Diphtheria, Leprosy, Paratuberculosis, Tuberculosis, Lupus, Botulism, gangrene, tetanus, impetigo, Rheumatic Fever, Scarlet Fever, sexually transmitted diseases, skin diseases (e.g., cellulitis, dermatocycoses), toxemia, urinary tract infections, wound infections. A polypeptide or polynucleotide of the present invention can be used to treat or detect any 30 of these symptoms or diseases. Moreover, parasitic agents causing disease or symptoms that can be treated or detected by a polynucleotide or polypeptide of the present invention include, but not 54 WO 01/30972 PCT/USOO/29690 limited to, the following families: Amebiasis, Babesiosis, Coccidiosis, Cryptosporidiosis, Dientamoebiasis, Dourine, Ectoparasitic, Giardiasis, Helminthiasis, Leishmaniasis, Theileriasis, Toxoplasmosis, Trypanosomiasis, and Trichomonas. These parasites can cause a variety of diseases or symptoms, including, but not limited to: Scabies, 5 Trombiculiasis, eye infections, intestinal disease (e.g., dysentery, giardiasis), liver disease, lung disease, opportunistic infections (e.g., AIDS related), Malaria, pregnancy complications, and toxoplasmosis. A polypeptide or polynucleotide of the present invention can be used to treat or detect any of these symptoms or diseases. 10 Preferably, treatment using a polypeptide or polynucleotide of the present invention could either be by administering an effective amount of a polypeptide to the patient, or by removing cells from the patient, supplying the cells with a polynucleotide of the present invention, and returning the engineered cells to the patient (ex vivo therapy). Moreover, the polypeptide or polynucleotide of the present invention can be 15 used as an antigen in a vaccine to raise an immune response against infectious disease. Regeneration A polynucleotide or polypeptide of the present invention can be used to differentiate, proliferate, and attract cells, leading to the regeneration of tissues (see, 20 Science, 276:59-87 (1997)). The regeneration of tissues could be used to repair, replace, or protect tissue damaged by congenital defects, trauma (wounds, burns, incisions, or ulcers), age, disease (e.g. osteoporosis, osteocarthritis, periodontal disease, liver failure), surgery, including cosmetic plastic surgery, fibrosis, reperfusion injury, or systemic cytokine damage. 25 Tissues that could be regenerated using the present invention include organs (e.g., pancreas, liver, intestine, kidney, skin, endothelium), muscle (smooth, skeletal or cardiac), vascular (including vascular endothelium), nervous, hematopoietic, and skeletal (bone, cartilage, tendon, and ligament) tissue. Preferably, regeneration occurs without or 30 decreased scarring. Regeneration also may include angiogenesis. Moreover, a polynucleotide or polypeptide of the present invention may increase regeneration of tissues difficult to heal. For example, increased tendon/ligament 55 WO 01/30972 PCT/USOO/29690 regeneration would quicken recovery time after damage. A polynucleotide or polypeptide of the present invention could also be used prophylactically in an effort to avoid damage. Specific diseases that could be treated include of tendinitis, carpal tunnel syndrome, and other tendon or ligament defects. A further example of tissue 5 regeneration of non-healing wounds includes pressure ulcers, ulcers associated with vascular insufficiency, surgical, and traumatic wounds. Similarly, nerve and brain tissue could also be regenerated by using a polynucleotide or polypeptide of the present invention to proliferate and differentiate 10 nerve cells. Diseases that could be treated using this method include central and peripheral nervous system diseases, neuropathies, or mechanical and traumatic disorders (e.g., spinal cord disorders, head trauma, cerebrovascular disease, and stroke). Specifically, diseases associated with peripheral nerve injuries, peripheral neuropathy (e.g., resulting from chemotherapy or other medical therapies), localized neuropathies, 15 and central nervous system diseases (e.g., Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, and Shy-Drager syndrome), could all be treated using the polynucleotide or polypeptide of the present invention. 20 Chemotaxis A polynucleotide or polypeptide of the present invention may have chemotaxis activity. A chemotaxic molecule attracts or mobilizes cells (e.g., monocytes, fibroblasts, neutrophils, T-cells, mast cells, eosinophils, epithelial and/or endothelial cells) to a particular site in the body, such as inflammation, infection, or site of hyperproliferation. 25 The mobilized cells can then fight off and/or heal the particular trauma or abnormality. A polynucleotide or polypeptide of the present invention may increase chemotaxic activity of particular cells. These chemotactic molecules can then be used to treat inflammation, infection, hyperproliferative disorders, or any immune system 30 disorder by increasing the number of cells targeted to a particular location in the body. For example, chemotaxic molecules can be used to treat wounds and other trauma to tissues by attracting immune cells to the injured location. Chemotactic molecules of the present invention can also attract fibroblasts, which can be used to treat wounds. 56 WO 01/30972 PCT/USOO/29690 It is also contemplated that a polynucleotide or polypeptide of the present invention may inhibit chemotactic activity. These molecules could also be used to treat disorders. Thus, a polynucleotide or polypeptide of the present invention could be used 5 as an inhibitor of chemotaxis. Binding Activity A polypeptide of the present invention may be used to screen for molecules that bind to the polypeptide or for molecules to which the polypeptide binds. The binding 10 of the polypeptide and the molecule may activate (i.e., an agonist), increase, inhibit (i.e., an antagonist), or decrease activity of the polypeptide or the molecule bound. Examples of such molecules include antibodies, oligonucleotides, proteins (e.g., receptors),or small molecules. 15 Preferably, the molecule is closely related to the natural ligand of the polypeptide, e.g., a fragment of the ligand, or a natural substrate, a ligand, a structural or functional mimetic (see, Coligan et al., Current Protocols in Immunology, 1(2), Chapter 5 (1991)). Similarly, the molecule can be closely related to the natural receptor to which the polypeptide binds, or at least, a fragment of the receptor capable of being bound by 20 the polypeptide (e.g., an active site). In either case, the molecule can be rationally designed using known techniques. Preferably, the screening for these molecules involves producing appropriate cells which express the polypeptide, either as a secreted protein or on the cell membrane. 25 Preferred cells include cells from mammals, yeast, Drosophila, or E. coli. Cells expressing the polypeptide (or cell membrane containing the expressed polypeptide) are then preferably contacted with a test compound potentially containing the molecule to observe binding, stimulation, or inhibition of activity of either the polypeptide or the molecule. 30 The assay may simply test binding of a candidate compound to the polypeptide, wherein binding is detected by a label, or in an assay involving competition with a 57 WO 01/30972 PCT/USOO/29690 labeled competitor. Further, the assay may test whether the candidate compound results in a signal generated by binding to the polypeptide. Alternatively, the assay can be carried out using cell-free preparations, 5 polypeptide/molecule affixed to a solid support, chemical libraries, or natural product mixtures. The assay may also simply comprise the steps of mixing a candidate compound with a solution containing a polypeptide, measuring polypeptide/molecule activity or binding, and comparing the polypeptide/molecule activity or binding to a standard. 10 Preferably, an ELISA assay can measure polypeptide level or activity in a sample (e.g., biological sample) using a monoclonal or polyclonal antibody. The antibody can measure polypeptide level or activity by either binding, directly or indirectly, to the polypeptide or by competing with the polypeptide for a substrate. 15 All of these above assays can be used as diagnostic or prognostic markers. The molecules discovered using these assays can be used to treat disease or to bring about a particular result in a patient (e.g., blood vessel growth) by activating or inhibiting the polypeptide/molecule. Moreover, the assays can discover agents which may inhibit or 20 enhance the production of the polypeptide from suitably manipulated cells or tissues. Therefore, the invention includes a method of identifying compounds which bind to a polypeptide of the invention comprising the steps of: (a) incubating a candidate binding compound with a polypeptide of the invention; and (b) determining if binding 25 has occurred. Moreover, the invention includes a method of identifying agonists/antagonists comprising the steps of: (a) incubating a candidate compound with a polypeptide of the invention, (b) assaying a biological activity, and (c) determining if a biological activity of the polypeptide has been altered. 30 Other Activities A polypeptide or polynucleotide of the present invention may also increase or decrease the differentiation or proliferation of embryonic stem cells, besides, as discussed above, hematopoietic lineage. 58 WO 01/30972 PCT/USOO/29690 A polypeptide or polynucleotide of the present invention may also be used to modulate mammalian characteristics, such as body height, weight, hair color, eye color, skin, percentage of adipose tissue, pigmentation, size, and shape (e.g., cosmetic surgery). 5 Similarly, a polypeptide or polynucleotide of the present invention may be used to modulate mammalian metabolism affecting catabolism, anabolism, processing, utilization, and storage of energy. A polypeptide or polynucleotide of the present invention may be used to change a 10 mammal's mental state or physical state by influencing biorhythms, circadian rhythms, depression (including depressive disorders), tendency for violence, tolerance for pain, the response to opiates and opioids, tolerance to opiates and opioids, withdrawal from opiates and opioids, reproductive capabilities (preferably by activin or inhibin-like activity), hormonal or endocrine levels, appetite, libido, memory, stress, or other cognitive 15 qualities. A polypeptide or polynucleotide of the present invention may also be used as a food additive or preservative, such as to increase or decrease storage capabilities, fat content, lipid, protein, carbohydrate, vitamins, minerals, cofactors or other nutritional 20 components. Other Preferred Embodiments Where a polynucleotide of the invention is down-regulated and exacerbates a pathological condition, such as psychosis or other neuropsychiatric disorders, the 25 expression of the polynucleotide can be increased or the level of the intact polypeptide product can be increased in order to treat, prevent, ameliorate, or modulate the pathological condition. This can be accomplished by, for example, administering a polynucleotide or polypeptide of the invention to the mammalian subject. 30 A polynucleotide of the invention can be administered to a mammalian subject by a recombinant expression vector comprising the polynucleotide. A mammalian subject can be a human, baboon, chimpanzee, macaque, cow, horse, sheep, pig, horse, dog, cat, rabbit, guinea pig, rat or mouse. Preferably, the recombinant vector comprises a 59 WO 01/30972 PCT/USOO/29690 polynucleotide shown in SEQ ID NOs: 1-19; 49-52; 57-72 and 107 or a polynucleotide which is at least 98% identical to a nucleic acid sequence shown in SEQ ID NOs: 1-19; 49-52; 57-72 and 107. Also, preferably, the recombinant vector comprises a variant polynucleotide that is at least 80%, 90%, or 95% identical to a polynucleotide comprising 5 SEQ ID NOs: 1-19; 49-52; 57-72 and 107. The administration of a polynucleotide or recombinant expression vector of the invention to a mammalian subject can be used to express a polynucleotide in said subject for the treatment of, for example, psychosis or other neuropsychiatric disorder. 10 Expression of a polynucleotide in target cells, including but not limited to cells of the striatum and nucleus accumbens, would effect greater production of the encoded polypeptide. In some cases where the encoded polypeptide is a nuclear protein, the regulation of other genes may be secondarily up- or down-regulated. 15 There are available to one skilled in the art multiple viral and non-viral methods suitable for introduction of a nucleic acid molecule into a target cell, as described above. In addition, a naked polynucleotide can be administered to target cells. Polynucleotides and recombinant expression vectors of the invention can be administered as a pharmaceutical composition. Such a composition comprises an effective amount of a 20 polynucleotide or recombinant expression vector, and a pharmaceutically acceptable formulation agent selected for suitability with the mode of administration. Suitable formulation materials preferably are non-toxic to recipients at the concentrations employed and can modify, maintain, or preserve, for example, the pH, osmolarity, viscosity, clarity, color, isotonicity, odor, sterility, stability, rate of dissolution or release, 25 adsorption, or penetration of the composition. See Remington 's Pharmaceutical Sciences

(

1 8 th Ed., A.R. Gennaro, ed., Mack Publishing Company 1990). The pharmaceutically active compounds (i.e., a polynucleotide or a vector) can be processed in accordance with conventional methods of pharmacy to produce medicinal 30 agents for administration to patients, including humans and other mammals. Thus, the pharmaceutical composition comprising a polynucleotide or a recombinant expression 60 WO 01/30972 PCT/USOO/29690 vector may be made up in a solid form (including granules, powders or suppositories) or in a liquid form (e.g., solutions, suspensions, or emulsions). The dosage regimen for treating a disease with a composition comprising a 5 polynucleotide or expression vector is based on a variety of factors, including the type or severity of the psychosis or other neuropsychiatric disorders, the age, weight, sex, medical condition of the patient, the route of administration, and the particular compound employed. Thus, the dosage regimen may vary widely, but can be determined routinely using standard methods. A typical dosage may range from about 0.1 mg/kg to about 100 10 mg/kg or more, depending on the factors mentioned above. The frequency of dosing will depend upon the pharmacokinetic parameters of the polynucleotide or vector in the formulation being used. Typically, a clinician will administer the composition until a dosage is reached that achieves the desired effect. The 15 composition may therefore be administered as a single dose, as two or more doses (which may or may not contain the same amount of the desired molecule) over time, or as a continuous infusion via implantation device or catheter. Further refinement of the appropriate dosage is routinely made by those of ordinary skill in the art and is within the ambit of tasks routinely performed by them. Appropriate dosages may be ascertained 20 through use of appropriate dose-response data. The cells of a mammalian subject may be transfected in vivo, ex vivo, or in vitro. Administration of a polynucleotide or a recombinant vector containing a polynucleotide to a target cell in vivo may be accomplished using any of a variety of techniques well 25 known to those skilled in the art. For example, U.S. Patent No. 5,672,344 describes an in vivo viral-mediated gene transfer system involving a recombinant neurotrophic HSV-1 vector. The above-described compositions of polynucleotides and recombinant vectors can be transfected in vivo by oral, buccal, parenteral, rectal, or topical administration as well as by inhalation spray. The term "parenteral" as used herein includes, subcutaneous, 30 intravenous, intramuscular, intrasternal, infusion techniques or intraperitoneally. 61 WO 01/30972 PCT/USOO/29690 While the nucleic acids and/or vectors of the invention can be administered as the sole active pharmaceutical agent, they can also be used in combination with one or more vectors of the invention or other agents. When administered as a combination, the therapeutic agents can be formulated as separate compositions that are given at the same 5 time or different times, or the therapeutic agents can be given as a single composition. - Another delivery system for polynucleotides of the invention is a "non-viral" delivery system. Techniques that have been used or proposed for gene therapy include DNA-ligand complexes, adenovirus-ligand-DNA complexes, direct injection of DNA, 10 CaPO 4 precipitation, gene gun techniques, electroporation, lipofection, and colloidal dispersion (Mulligan, R., (1993) Science, 260 (5110):926-32 (1993)). Any of these methods are widely available to one skilled in the art and would be suitable for use in the present invention. Other suitable methods are available to one skilled in the art, and it is to be understood that the present invention may be accomplished using any of the 15 available methods of transfection. Several such methodologies have been utilized by those skilled in the art with varying success (Mulligan, R., (1993) Science, 260 (5110):926-32 (1993)). Where a polynucleotide of the invention is up-regulated and exacerbates a 20 pathological condition in a mammalian subject, such as psychosis or other neuropsychiatric disorders, the expression of the polynucleotide can be blocked or reduced or the level of the intact polypeptide product can be reduced in order to treat, prevent, ameliorate, or modulate the pathological condition. This can be accomplished by, for example, the use of antisense oligonucleotides or ribozymes. Alternatively, drugs 25 or antibodies that bind to and inactivate the polypeptide product can be used. 62 WO 01/30972 PCT/USOO/29690 Antisense oligonucleotides are nucleotide sequences which are complementary to a specific DNA or RNA sequence. Once introduced into a cell, the complementary nucleotides combine with natural sequences produced by the cell to form complexes and 5 block either transcription or translation. Preferably, an antisense oligonucleotide is at least 11 nucleotides in length, but can be at least 12, 15, 20, 25, 30, 35, 40, 45, or 50 or more nucleotides long. Longer sequences also can be used. Antisense oligonucleotide molecules can be provided in a DNA construct and introduced into a cell as described above to decrease the level of gene products of the invention in the cell. 10 Antisense oligonucleotides can be deoxyribonucleotides, ribonucleotides, or a combination of both. Oligonucleotides can be synthesized manually or by an automated synthesizer, by covalently linking the 5' end of one nucleotide with the 3' end of another nucleotide with non-phosphodiester internucleotide linkages such alkylphosphonates, 15 phosphorothioates, phosphorodithioates, alkylphosphonothioates, alkylphosphonates, phosphoramidates, phosphate esters, carbamates, acetamidate, carboxymethyl esters, carbonates, and phosphate triesters. See Brown, (1994) Meth. Mo. Bio., 20:1-8; Sonveaux, (1994) Meth. Mo. Bio., 26:1-72; Uhlmann et al., (1990) Chem. Rev., 90:543 583. 20 Modifications of gene expression can be obtained by designing antisense oligonucleotides which will form duplexes to the control, 5', or regulatory regions of a gene of the invention. Oligonucleotides derived from the transcription initiation site, e.g., between positions -10 and +10 from the start site, are preferred. Similarly, inhibition can 25 be achieved using "triple helix" base-pairing methodology. Triple helix pairing is useful because it causes inhibition of the ability of the double helix to open sufficiently for the binding of polymerases, transcription factors, or chaperons. Therapeutic advances using triplex DNA have been described in the literature (e.g., Gee et al., in Huber & Carr, MOLECULAR AND IMMUNOLOGIC APPROACHES, Futura Publishing Co., Mt. Kisco, N.Y., 30 1994). An antisense oligonucleotide also can be designed to block translation of mRNA by preventing the transcript from binding to ribosomes. 63 WO 01/30972 PCT/USOO/29690 Precise complementarity is not required for successful complex formation between an antisense oligonucleotide and the complementary sequence of a polynucleotide. Antisense oligonucleotides which comprise, for example, 2, 3, 4, or 5 or 5 more stretches of contiguous nucleotides which are precisely complementary to a polynucleotide, each separated by a stretch of contiguous nucleotides which are not complementary to adjacent nucleotides, can provide sufficient targeting specificity for mRNA. Preferably, each stretch of complementary contiguous nucleotides is at least 4, 5, 6, 7, or 8 or more nucleotides in length. Non-complementary intervening sequences 10 are preferably 1, 2, 3, or 4 nucleotides in length. One skilled in the art can easily use the calculated melting point of an antisense-sense pair to determine the degree of mismatching which will be tolerated between a particular antisense oligonucleotide and a particular polynucleotide sequence. 15 Antisense oligonucleotides can be modified without affecting their ability to hybridize to a polynucleotide of the invention. These modifications can be internal or at one or both ends of the antisense molecule. For example, internucleoside phosphate linkages can be modified by adding cholesteryl or diamine moieties with varying numbers of carbon residues between the amino groups and terminal ribose. Modified 20 bases and/or sugars, such as arabinose instead of ribose, or a 3', 5'-substituted oligonucleotide in which the 3' hydroxyl group or the 5' phosphate group are substituted, also can be employed in a modified antisense oligonucleotide. These modified oligonucleotides can be prepared by methods well known in the art. See, e.g., Agrawal et al., (1992) Trends Biotechnol., 10:152-158; Uhlmann et al., (1990) Chem. Rev., 90:543 25 584; Uhlmann et al., (1987) Tetrahedron. Lett., 215:3539-3542. 64 WO 01/30972 PCT/USOO/29690 Ribozymes are RNA molecules with catalytic activity. See, e.g., Cech, (1987) Science, 236:1532-1539; Cech, (1990) Ann. Rev. Biochem., 59:543-568; Cech, (1992) Curr. Opin. Struct. Biol., 2:605-609; Couture & Stinchcomb, (1996) Trends Genet., 12:510-515. Ribozymes can be used to inhibit gene function by cleaving an RNA 5 sequence, as is known in the art (e.g., Haseloff et al., U.S. Patent 5,641,673). The mechanism of ribozyme action involves sequence-specific hybridization of the ribozyme molecule to complementary target RNA, followed by endonucleolytic cleavage. Examples include engineered hammerhead motif ribozyme molecules that can specifically and efficiently catalyze endonucleolytic cleavage of specific nucleotide 10 sequences. The coding sequence of a polynucleotide of the invention can be used to generate ribozymes which will specifically bind to mRNA transcribed from the polynucleotide. Methods of designing and constructing ribozymes which can cleave RNA molecules in 15 trans in a highly sequence specific manner have been developed and described in the art (see Haseloff et al. (1988) Nature, 334:585-591). For example, the cleavage activity of ribozymes can be targeted to specific RNAs by engineering a discrete "hybridization" region into the ribozyme. The hybridization region contains a sequence complementary to the target RNA and thus specifically hybridizes with the target (see, e.g., Gerlach et al., 20 EP 321,201). Specific ribozyme cleavage sites within a RNA target can be identified by scanning the target molecule for ribozyme cleavage sites which include the following sequences: GUA, GUU, and GUC. Once identified, short RNA sequences of between 15 25 and 20 ribonucleotides corresponding to the region of the target RNA containing the cleavage site can be evaluated for secondary structural features which may render the target inoperable. Suitability of candidate RNA targets also can be evaluated by testing accessibility to hybridization with complementary oligonucleotides using ribonuclease protection assays. The nucleotide sequences shown in SEQ ID NOs: 1-19; 49-52; 57-72 30 and 107 and their complements provide sources of suitable hybridization region sequences. Longer complementary sequences can be used to increase the affinity of the 65 WO 01/30972 PCT/USOO/29690 hybridization sequence for the target. The hybridizing and cleavage regions of the ribozyme can be integrally related such that upon hybridizing to the target RNA through the complementary regions, the catalytic region of the ribozyme can cleave the target. 5 Ribozymes can be introduced into cells as part of a DNA construct. Mechanical methods, such as microinjection, liposome-mediated transfection, electroporation, or calcium phosphate precipitation, can be used to introduce a ribozyme-containing DNA construct into cells in which it is desired to decrease polynucleotide expression. Alternatively, if it is desired that the cells stably retain the DNA construct, the construct 10 can be supplied on a plasmid and maintained as a separate element or integrated into the genome of the cells, as is known in the art. A ribozyme-encoding DNA construct can include transcriptional regulatory elements, such as a promoter element, an enhancer or UAS element, and a transcriptional terminator signal, for controlling transcription of ribozymes in the cells. 15 As taught in Haseloff et al., U.S. Patent 5,641,673, ribozymes can be engineered so that ribozyme expression will occur in response to factors which induce expression of a target gene. Ribozymes also can be engineered to provide an additional level of regulation, so that destruction of mRNA occurs only when both a ribozyme and a target 20 gene are induced in the cells. Production of Diagnostic Tests Pathological conditions or susceptibility to pathological conditions, such as psychosis or other neuropsychiatric disorders, can be diagnosed using methods of the 25 invention. Testing for expression of a polynucleotide of the invention or for the presence of the polynucleotide product can correlate with the severity of the condition and can also indicate appropriate treatment. For example, the presence or absence of a mutation in a polynucleotide of the invention can be determined and a pathological condition or a susceptibility to a pathological condition is diagnosed based on the presence or absence 30 of the mutation. Further, an alteration in expression of a polypeptide encoded by a polynucleotide of the invention can be detected, where the presence of an alteration in 66 WO 01/30972 PCT/USOO/29690 expression of the polypeptide is indicative of the pathological condition or susceptibility to the pathological condition. The alteration in expression can be an increase in the amount of expression or a decrease in the amount of expression. 5 As an additional method of diagnosis, a first biological sample from a patient suspected of having a pathological condition, such as psychosis or addiction-related behavior, is obtained along with a second sample from a suitable comparable control source. A biological sample can comprise saliva, blood, cerebrospinal fluid, amniotic fluid, urine, feces, or tissue, such as gastrointestinal tissue. A suitable control source can 10 be obtained from one or more mammalian subjects that do not have the pathological condition. For example, the average concentrations and distribution of a polynucleotide or polypeptide of the invention can be determined from biological samples taken from a representative population of mammalian subjects, wherein the mammalian subjects are the same species as the subject from which the test sample was obtained. The amount of 15 at least one polypeptide encoded by a polynucleotide of the invention is determined in the first and second sample. The amounts of the polypeptide in the first and second samples are compared. A patient is diagnosed as having a pathological condition if the amount of the polypeptide in the first sample falls in the range of samples taken from a representative group of patients with the pathological condition. 20 Other preferred embodiments of the claimed invention include an isolated nucleic acid molecule comprising a nucleotide sequence which is at least 80%, preferably at least 85%, more preferably at least 90%, most preferably at least 95% identical to a sequence of at least about 50 contiguous nucleotides in the nucleotide sequence of SEQ ID NOs: 1 25 19; 49-52; 57-72 and 107. Also preferred is a nucleic acid molecule wherein said sequence of contiguous nucleotides is included in the nucleotide sequence of SEQ ID NOs: 1-19; 49-52; 57-72 and 107 in the range of positions beginning with the nucleotide at about the position of 30 the 5' nucleotide of the clone sequence and ending with the nucleotide at about the position of the 3' nucleotide of the clone sequence. 67 WO 01/30972 PCT/USOO/29690 Also preferred is a nucleic acid molecule wherein said sequence of contiguous nucleotides is included in the nucleotide sequence of SEQ ID NOs: 1-19; 49-52; 57-72 and 107 in the range of positions beginning with the nucleotide at about the position of the 5' nucleotide of the start codon and ending with the nucleotide at about the position of 5 the 3' nucleotide of the clone sequence as defined for SEQ ID NOs: 1-19; 49-52; 57-72 and 107. Similarly preferred is a nucleic acid molecule wherein said sequence of contiguous nucleotides is included in the nucleotide sequence of SEQ ID NOs: 1-19; 49 10 52; 57-72 and 107 in the range of positions beginning with the nucleotide at about the position of the 5' nucleotide of the first amino acid of the signal peptide and ending with the nucleotide at about the position of the 3' nucleotide of the clone sequence as defined for SEQ ID NOs: 1-19; 49-52; 57-72 and 107. 15 Also preferred is an isolated nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to a sequence of at least about 150 contiguous nucleotides in the nucleotide sequence of SEQ ID NOs: 1-19; 49-52; 57-72 and 107. Further preferred is an isolated nucleic acid molecule comprising a nucleotide 20 sequence which is at least 95% identical to a sequence of at least about 500 contiguous nucleotides in the nucleotide sequence of SEQ ID NOs: 1-19; 49-52; 57-72 and 107. A further preferred embodiment is a nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to the nucleotide sequence of SEQ ID 25 NOs: 1-19; 49-52; 57-72 and 107 beginning with the nucleotide at about the position of the 5' nucleotide of the first amino acid of the signal peptide and ending with the nucleotide at about the position of the 3' nucleotide of the clone sequence as defined for SEQ ID NOs: 1-19; 49-52; 57-72 and 107. 30 A further preferred embodiment is an isolated nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to the complete nucleotide sequence of SEQ ID NOs: 1-19; 49-52; 57-72 and 107. In another embodiment, the present invention provides a method for detecting in a biological sample a nucleic acid molecule 68 WO 01/30972 PCT/USOO/29690 comprising a nucleotide sequence which is at least 95% identical to a complete nucleotide sequence chosen from the group consisting of SEQ ID NOs: 1-19; 49-52; 57 72 and 107, which method comprises the steps of comparing a nucleotide sequence of at least one nucleic acid molecule in said sample with a sequence selected from said group 5 and determining whether the sequence of said nucleic acid molecule in said sample is at least 95% identical to said selected sequence. Also preferred is an isolated nucleic acid molecule which hybridizes under 10 stringent hybridization conditions to a nucleic acid molecule, wherein said nucleic acid molecule which hybridizes does not hybridize under stringent hybridization conditions to a nucleic acid molecule having a nucleotide sequence consisting of only A residues or of only T residues. 15 A further preferred embodiment is a method for detecting in a biological sample a nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to a sequence of at least 50 contiguous nucleotides in a sequence selected from the group consisting of: a nucleotide sequence of SEQ ID NOs: 1-19; 49-52; 57-72 and 107, which method comprises the steps of comparing a nucleotide sequence of at least one nucleic 20 acid molecule in said sample with a sequence selected from said group and determining whether the sequence of said nucleic acid molecule in said sample is at least 95% identical to said selected sequence. Also preferred is the above method wherein said step of comparing sequences 25 comprises determining the extent of nucleic acid hybridization between nucleic acid molecules in said sample and a nucleic acid molecule comprising said sequence selected from said group. Similarly, also preferred is the above method wherein said step of comparing sequences is performed by comparing the nucleotide sequence determined from a nucleic acid molecule in said sample with said sequence selected from said group. 30 The nucleic acid molecules can comprise DNA molecules or RNA molecules. 69 WO 01/30972 PCT/USOO/29690 A further preferred embodiment is a method for identifying the species, tissue or cell type of a biological sample which method comprises a step of detecting nucleic acid molecules in said sample, if any, comprising a nucleotide sequence that is at least 95% identical to a sequence of at least 50 contiguous nucleotides in a sequence selected from 5 the group consisting of: a nucleotide sequence of SEQ ID NOs: 1-19; 49-52; 57-72 and 107. Also preferred is a method for diagnosing in a subject a pathological condition associated with abnormal structure or expression of a gene, which method comprises a 10 step of detecting in a biological sample obtained from said subject nucleic acid molecules, if any, comprising a nucleotide sequence that is at least 95% identical to a sequence of at least 50 contiguous nucleotides in a sequence selected from the group consisting of. a nucleotide sequence of SEQ ID NOs: 1-19; 49-52; 57-72 and 107. 15 The method for diagnosing a pathological condition can comprise a step of detecting nucleic acid molecules comprising a nucleotide sequence in a panel of at least two nucleotide sequences, wherein at least one sequence in said panel is at least 95% identical to a sequence of at least 50 contiguous nucleotides in a sequence selected from said group. 20 Also preferred is a composition of matter comprising isolated nucleic acid molecules wherein the nucleotide sequences of said nucleic acid molecules comprise a panel of at least two nucleotide sequences, wherein at least one sequence in said panel is at least 95% identical to a sequence of at least 50 contiguous nucleotides in a sequence 25 selected from the group consisting of: a nucleotide sequence of SEQ ID NOs: 1-19; 49 52; 57-72 and 107. The nucleic acid molecules can comprise DNA molecules or RNA molecules. Also preferred is an isolated polypeptide comprising an amino acid sequence at 30 least 90% identical to a sequence of at least about 10 contiguous amino acids in an amino acid sequence translated from SEQ ID NOs: 1-19; 49-52; 57-72 and 107. 70 WO 01/30972 PCT/USOO/29690 Also preferred is a polypeptide, wherein said sequence of contiguous amino acids is included in acids in an amino acid sequence translated from SEQ ID NOs: 1-19; 49-52; 57-72 and 107, in the range of positions beginning with the residue at about the position of the first amino acid of the secreted portion and ending with the residue at 5 about the last amino acid of the open reading frame. Also preferred is an isolated polypeptide comprising an amino acid sequence at least 95% identical to a sequence of at least about 30 contiguous amino acids in an amino acid sequence translated from SEQ ID NOs: 1-19; 49-52; 57-72 and 107. 10 Further preferred is an isolated polypeptide comprising an amino acid sequence at least 95% identical to a sequence of at least about 100 contiguous amino acids in an amino acid sequence translated from SEQ ID NOs: 1-19; 49-52; 57-72 and 107. 15 Further preferred is an isolated polypeptide comprising an amino acid sequence at least 95% identical to acids in an amino acid sequence translated from SEQ ID NOs: 1 19; 49-52; 57-72 and 107. Further preferred is a method for detecting in a biological sample a polypeptide 20 comprising an amino acid sequence which is at least 90% identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the group consisting of amino acid sequences translated from SEQ ID NOs: 1-19; 49-52; 57-72 and 107, which method comprises a step of comparing an amino acid sequence of at least one polypeptide molecule in said sample with a sequence selected from said group and 25 determining whether the sequence of said polypeptide molecule in said sample is at least 90% identical to said sequence of at least 10 contiguous amino acids. Also preferred is the above method wherein said step of comparing an amino acid sequence of at least one polypeptide molecule in said sample with a sequence selected 30 from said group comprises determining the extent of specific binding of polypeptides in said sample to an antibody which binds specifically to a polypeptide comprising an amino acid sequence that is at least 90% identical to a sequence of at least 10 contiguous 71 WO 01/30972 PCT/USOO/29690 amino acids in a sequence selected from the group consisting of amino acid sequences translated from SEQ ID NOs: 1-19; 49-52; 57-72 and 107. Also preferred is the above method wherein said step of comparing sequences is 5 performed by comparing the amino acid sequence determined from a polypeptide molecule in said sample with said sequence selected from said group. Also preferred is a method for identifying the species, tissue or cell type of a biological sample which method comprises a step of detecting polypeptide molecules in 10 said sample, if any, comprising an amino acid sequence that is at least 90% identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the group consisting of amino acid sequences translated from SEQ ID NOs: 1-19; 49-52; 57-72 and 107. 15 Also preferred is the above method for identifying the species, tissue or cell type of a biological sample, which method comprises a step of detecting polypeptide molecules comprising an amino acid sequence in a panel of at least two amino acid sequences, wherein at least one sequence in said panel is at least 90% identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the above 20 group. Also preferred is a method for diagnosing in a subject a pathological condition associated with abnormal structure or expression of a gene, which method comprises a step of detecting in a biological sample obtained from said subject polypeptide molecules 25 comprising an amino acid sequence in a panel of at least two amino acid sequences, wherein at least one sequence in said panel is at least 90% identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the group consisting of amino acid sequences translated from SEQ ID NOs: 1-19; 49-52; 57-72 and 107. 30 In any of these methods, the step of detecting said polypeptide molecules includes using an antibody. 72 WO 01/30972 PCT/USOO/29690 Also preferred is an isolated nucleic acid molecule comprising a nucleotide sequence which is at least 95% identical to a nucleotide sequence encoding a polypeptide wherein said polypeptide comprises an amino acid sequence that is at least 90% identical to a sequence of at least 10 contiguous amino acids in a sequence selected from the group 5 consisting of amino acid sequences translated from SEQ ID NOs: 1-19; 49-52; 57-72 and 107. Also preferred is an isolated nucleic acid molecule, wherein said nucleotide sequence encoding a polypeptide has been optimized for expression of said polypeptide 10 in a prokaryotic host. Also preferred is an isolated nucleic acid molecule, wherein said nucleotide sequence encodes a polypeptide comprising an amino acid sequence selected from the group consisting of amino acid sequences translated from SEQ ID NOs: 1-19; 49-52; 57 15 72 and 107. Further preferred is a method of making a recombinant vector comprising inserting any of the above isolated nucleic acid molecule into a vector. Also preferred is the recombinant vector produced by this method. Also preferred is a method of making a 20 recombinant host cell comprising introducing the vector into a host cell, as well as the recombinant host cell produced by this method. Also preferred is a method of making an isolated polypeptide comprising culturing this recombinant host cell under conditions such that said polypeptide is 25 expressed and recovering said polypeptide. Also preferred is this method of making an isolated polypeptide, wherein said recombinant host cell is a eukaryotic cell and said polypeptide is a secreted portion of a human secreted protein comprising an amino acid sequence selected from the group consisting of amino acid sequences translated from SEQ ID NOs: 1-19; 49-52; 57-72 and 107. The isolated polypeptide produced by this 30 method is also preferred. Also preferred is a method of treatment of an individual in need of an increased level of a secreted protein activity, which method comprises administering to such an 73 WO 01/30972 PCT/USOO/29690 individual a pharmaceutical composition comprising an amount of an isolated polypeptide, polynucleotide, or antibody of the claimed invention effective to increase the level of said protein activity in said individual. 5 The present invention also includes a diagnostic system, preferably in kit form, for assaying for the presence of the polypeptide of the present invention in a body sample, such brain tissue, cell suspensions or tissue sections, or body fluid samples such as CSF, blood, plasma or serum, where it is desirable to detect the presence, and preferably the amount, of the polypeptide of this invention in the sample according to the diagnostic 10 methods described herein. In a related embodiment, a nucleic acid molecule can be used as a probe (an oligonucleotide) to detect the presence of a polynucleotide of the present invention, or a gene corresponding to a polynucleotide of the present invention, or a mRNA in a cell that 15 is diagnostic for the presence or expression of a polypeptide of the present invention in the cell. The nucleic acid molecule probes can be of a variety of lengths from at least about 10, suitably about 10 to about 5000 nucleotides long, although they will typically be about 20 to 500 nucleotides in length. Hybridization methods are extremely well known in the art and will not be described further here. 20 In a related embodiment, detection of genes corresponding to the polynucleotides of the present invention can be conducted by primer extension reactions such as the polymerase chain reaction (PCR). To that end, PCR primers are utilized in pairs, as is well known, based on the nucleotide sequence of the gene to be detected. Preferably the 25 nucleotide sequence is a portion of the nucleotide sequence of a polynucleotide of the present invention. Particularly preferred PCR primers can be derived from any portion of a DNA sequence encoding a polypeptide of the present invention, but are preferentially from regions which are not conserved in other cellular proteins. 30 Preferred PCR primer pairs useful for detecting the genes corresponding to the polynucleotides of the present invention and expression of these genes are described in 74 WO 01/30972 PCT/USOO/29690 the Examples, including the corresponding Tables. Nucleotide primers from the corresponding region of the polypeptides of the present invention described herein are readily prepared and used as PCR primers for detection of the presence or expression of the corresponding gene in any of a variety of tissues. 5 The diagnostic system includes, in an amount sufficient to perform at least one assay, a subject polypeptide of the present invention, a subject antibody or monoclonal antibody, and/or a subject nucleic acid molecule probe of the present invention, as a separately packaged reagent. 10 In another embodiment, a diagnostic system, preferably in kit form, is contemplated for assaying for the presence of the polypeptide of the present invention or an antibody immunoreactive with the polypeptide of the present invention in a body fluid sample such as for monitoring the fate of therapeutically administered the polypeptide of 15 the present invention or an antibody immunoreactive with the polypeptide of the present invention. The system includes, in an amount sufficient for at least one assay, a polypeptide of the present invention and/or a subject antibody as a separately packaged immunochemical reagent. 20 Instructions for use of the packaged reagent(s) are also typically included. As used herein, the term "package" refers to a solid matrix or material such as glass, plastic (e.g., polyethylene, polypropylene or polycarbonate), paper, foil and the like capable of holding within fixed limits a polypeptide, polyclonal antibody or monoclonal 25 antibody of the present invention. Thus, for example, a package can be a glass vial used to contain milligram quantities of a contemplated polypeptide or antibody or it can be a microtiter plate well to which microgram quantities of a contemplated polypeptide or antibody have been operatively affixed, i.e., linked so as to be capable of being immunologically bound by an antibody or antigen, respectively. 30 75 WO 01/30972 PCT/USOO/29690 "Instructions for use" typically include a tangible expression describing the reagent concentration or at least one assay method parameter such as the relative amounts of reagent and sample to be admixed, maintenance time periods for reagent/ sample admixtures, temperature, buffer conditions and the like. 5 A diagnostic system of the present invention preferably also includes a label or indicating means capable of signaling the formation of an immunocomplex containing a polypeptide or antibody molecule of the present invention. 10 The word "complex" as used herein refers to the product of a specific binding reaction such as an antibody-antigen or receptor-ligand reaction. Exemplary complexes are immunoreaction products. As used herein, the terms "label" and "indicating means" in their various 15 grammatical forms refer to single atoms and molecules that are either directly or indirectly involved in the production of a detectable signal to indicate the presence of a complex. Any label or indicating means can be linked to or incorporated in an expressed protein, polypeptide, or antibody molecule that is part of an antibody or monoclonal antibody composition of the present invention, or used separately, and those atoms or 20 molecules can be used alone or in conjunction with additional reagents. Such labels are themselves well-known in clinical diagnostic chemistry and constitute a part of this invention only insofar as they are utilized with otherwise novel proteins methods and/or systems. 25 The labeling means can be a fluorescent labeling agent that chemically binds to antibodies or antigens without denaturing them to form a fluorochrome (dye) that is a useful immunofluorescent tracer. Suitable fluorescent labeling agents are fluorochromes such as fluorescein isocyanate (FIC), fluorescein isothiocyante (FITC), 5-dimethylamine 1-naphthalenesulfonyl chloride (DANSC), tetramethylrhodamine isothiocyanate 30 (TRITC), lissamine, rhodamine 8200 sulphonyl chloride (RB 200 SC) and the like. A description of immunofluorescence analysis techniques is found in DeLuca, 76 WO 01/30972 PCT/USOO/29690 "Immunofluorescence Analysis", in Antibody As a Tool, Marchalonis, et al., Eds., John Wiley & Sons, Ltd., pp. 189-231 (1982), which is incorporated herein by reference. Other suitable labeling agents are known to those skilled in the art. 5 In preferred embodiments, the indicating group is an enzyme, such as horseradish peroxidase (HRP), glucose oxidase, or the like. In such cases where the principal indicating group is an enzyme such as HRP or glucose oxidase, additional reagents are required to visualize the fact that a receptor-ligand complex (immunoreactant) has formed. Such additional reagents for HRP include hydrogen peroxide and an oxidation 10 dye precursor such as diaminobenzidine. An additional reagent useful with glucose oxidase is 2,2'-amino-di-(3-ethyl-benzthiazoline-G-sulfonic acid) (ABTS). Radioactive elements are also useful labeling agents and are used illustratively herein. An exemplary radiolabeling agent is a radioactive element that produces gamma 15 ray emissions. Elements which themselves emit gamma rays, such as 14, 1251, 1281, 1321 and 51 Cr represent one class of gamma ray emission-producing radioactive element indicating groups. Particularly preferred is 125. Another group of useful labeling means are those elements such as "IC, " 8 F, 1s0 and "N which themselves emit positrons. The positrons so emitted produce gamma rays upon encounters with electrons present in the 20 animal's body. Also useful is a beta emitter, such 1" indium or 3H. The linking of labels, i.e., labeling of, polypeptides and proteins is well known in the art. For instance, antibody molecules produced by a hybridoma can be labeled by metabolic incorporation of radioisotope-containing amino acids provided as a component 25 in the culture medium (see, e.g., Galfre et al., Meth. Enzymol., 73:3-46 (1981)). The techniques of protein conjugation or coupling through activated functional groups are particularly applicable (see, e.g.,Aurameas, et al., Scand. J. Immunol., Vol. 8 Suppl. 7:7 23 (1978); Rodwell et al., Biotech., 3:889-894 (1984); and U.S. Pat. No. 4,493,795). 30 The diagnostic systems can also include, preferably as a separate package, a specific binding agent. A "specific binding agent" is a molecular entity capable of 77 WO 01/30972 PCT/USOO/29690 selectively binding a reagent species of the present invention or a complex containing such a species, but is not itself a polypeptide or antibody molecule composition of the present invention. Exemplary specific binding agents are second antibody molecules, complement proteins or fragments thereof, S. aureus protein A, and the like. Preferably 5 the specific binding agent binds the reagent species when that species is present as part of a complex. In preferred embodiments, the specific binding agent is labeled. However, when the diagnostic system includes a specific binding agent that is not labeled, the agent is 10 typically used as an amplifying means or reagent. In these embodiments, the labeled specific binding agent is capable of specifically binding the amplifying means when the amplifying means is bound to a reagent species-containing complex. The diagnostic kits of the present invention can be used in an "ELISA" format to 15 detect the quantity of the polypeptide of the present invention in a sample. "ELISA" refers to an enzyme-linked immunosorbent assay that employs an antibody or antigen bound to a solid phase and an enzyme-antigen or enzyme-antibody conjugate to detect and quantify the amount of an antigen present in a sample. A description of the ELISA technique is found in Sites et al., Basic and Clinical Immunology, 4th Ed., Lange Medical 20 Publications, Los Altos, CA (1982) and in U.S. Patents No. 3,654,090; No. 3,850,752; and No. 4,016,043, which are all incorporated herein by reference. Thus, in some embodiments, an polypeptide of the present invention, an antibody or a monoclonal antibody of the present invention can be affixed to a solid matrix to form 25 a solid support that comprises a package in the subject diagnostic systems. A reagent is typically affixed to a solid matrix by adsorption from an aqueous medium although other modes of affixation applicable to proteins and polypeptides can be used that are well known to those skilled in the art. Exemplary adsorption methods 30 are described herein. 78 WO 01/30972 PCT/USOO/29690 Useful solid matrices are also well known in the art. Such materials are water insoluble and include the cross-linked dextran available under the trademark SEPHADEX from Pharmacia Fine Chemicals (Piscataway, NJ); agarose; beads of polystyrene beads about 1 micron (tm) to about 5 millimeters (mm) in diameter available 5 from several suppliers, e.g., Abbott Laboratories of North Chicago, IL; polyvinyl chloride, polystyrene, cross-linked polyacrylamide, nitrocellulose- or nylon-based webs such as sheets, strips or paddles; or tubes, plates or the wells of a microtiter plate such as those made from polystyrene or polyvinylchloride. 10 The reagent species, labeled specific binding agent or amplifying reagent of any diagnostic system described herein can be provided in solution, as a liquid dispersion or as a substantially dry power, e.g., in lyophilized form. Where the indicating means is an enzyme, the enzyme's substrate can also be provided in a separate package of a system. A solid support such as the before-described microtiter plate and one or more buffers can 15 also be included as separately packaged elements in this diagnostic assay system. The packaging materials discussed herein in relation to diagnostic systems are those customarily utilized in diagnostic systems. 20 Having generally described the invention, the same will be more readily understood by reference to the following examples, which are provided by way of illustration and are not intended as limiting. 25 EXAMPLE 1 Identification and Characterization of Polynucleotides Regulated by Neuroleptic Drugs Male C57B1/6J mice (20-28 g) were housed in groups of four on a standard 12/12 30 hour light-dark cycle with ad libitum access to standard laboratory chow and tap water. For the experimental paradigms, mice were divided into groups of 25 and subjected to the following treatments: 79 WO 01/30972 PCT/USOO/29690 Control groups: Mice received a single injection of sterile saline (0.1 ml volume), or no injection, and were sacrificed after 45 minutes. Acute neuroleptic treatment: Mice received a single intraperitoneal injection of the atypical neuroleptic clozapine (7.5 mg/kg). Animals were sacrificed after 45 minutes. 5 Chronic neuroleptic treatment: Mice received daily subcutaneous injections of clozapine (7.5 mg/kg) for time periods of 5 days to 2 weeks. All animals were sacrificed in their cages with CO 2 at the indicated times. Brains were rapidly removed and placed on ice. The striatum, including the nucleus accumbens, 10 were dissected out and placed in ice-cold phosphate-buffered saline. Isolated RNA was analyzed using a method of simultaneous sequence-specific identification of mRNAs known as TOGA (TOtal Gene expression Analysis) described in Sutcliffe et al. Proc. Natl. Acad. Sci. USA, 97(5):1976-1981 (2000); International 15 published application WO 026406; U.S. Patent No. 5,459,037; U.S. Patent No. 5,807,680; U.S. Patent No. 6,030,784; U.S. Patent No. 6,096,503 and U.S. Patent 6,110,680, hereby incorporated herein by reference. Preferably, prior to the application of the TOGA technique, the isolated RNA was enriched to form a starting polyA containing mRNA population by methods known in the art. In a preferred embodiment, 20 the TOGA method further comprised an additional PCR step performed using four 5' PCR primers in four separate reactions and cDNA templates prepared from a population of antisense cRNAs. A final PCR step that used 256 5' PCR primers in separate reactions produced PCR products that were cDNA fragments that corresponded to the 3'-region of the starting mRNA population. The produced PCR products were then identified by: a) 25 the initial 5' sequence comprising the sequence remainder of the recognition site of the restriction endonuclease used to cut and isolate the 3' region plus the sequence of the preferably four parsing bases immediately 3' to the remainder of the recognition site, preferably the sequence of the entire fragment, and b) the length of the fragment. These two parameters, sequence and fragment length, were used to compare the obtained PCR 30 products to a database of known polynucleotide sequences. Since the length of the obtained PCR products includes known vector sequences at the 5' and 3' ends of the 80 WO 01/30972 PCT/USOO/29690 insert, the sequence of the insert provided in the sequence listing is shorter than the fragment length that forms part of the digital address. The method yields Digital Sequence Tags (DSTs), that is, polynucleotides that are 5 expressed sequence tags of the 3' end of mRNAs. DSTs that showed changes in relative levels as a result of clozapine treatment were selected for further study. The intensities of the laser-induced fluorescence of the labeled PCR products were compared across samples isolated from the striatum/nucleus accumbens of mice treated with clozapine for 45 minutes, 7 hours, 5 days, 12 days, or 14 days. 10 In general, double-stranded cDNA is generated from poly(A)-enriched cytoplasmic RNA extracted from the tissue samples of interest using an equimolar mixture or set of all 48 5'-biotinylated anchor primers to initiate reverse transcription. One such suitable set is G-A-A-T-T-C-A-A-C-T-G-G-A-A-G-C-G-G-C-C-G-C-A-G-G 15 A-A-T-T-T-T-T-T-T-T-T-T-T-T-T-T-T-T-T-T-V-N-N (SEQ ID NO: 20), where V is A, C or G and N is A, C, G or T. One member of this mixture of 48 anchor primers initiates synthesis at a fixed position at the 3' end of all copies of each mRNA species in the sample, thereby defining a 3' endpoint for each species, resulting in biotinylated double stranded cDNA. 20 Each biotinylated double stranded cDNA sample was cleaved with the restriction endonuclease MsgI, which recognizes the sequence CCGG. The resulting fragments of cDNA corresponding to the 3' region of the starting mRNA were then isolated by capture of the biotinylated cDNA fragments on a streptavidin-coated substrate. Suitable 25 streptavidin-coated substrates include microtitre plates, PCR tubes, polystyrene beads, paramagnetic polymer beads and paramagnetic porous glass particles. A preferred streptavidin-coated substrate is a suspension of paramagnetic polymer beads (Dynal, Inc., Lake Success, NY). 30 After washing the streptavidin-coated substrate and captured biotinylated cDNA fragments, the cDNA fragment product was released by digestion with NotI, which 81 WO 01/30972 PCT/USOO/29690 cleaves at an 8-nucleotide sequence within the anchor primers but rarely within the mRNA-derived portion of the cDNAs. The 3' MspI-NotI fragments, which are of uniform length for each mRNA species, were directionally ligated into ClaI- NotI cleaved plasmid pBC SK* (Stratagene, La Jolla, CA) in an antisense orientation with 5 respect to the vector's T3 promoter, and the product used to transform Escherichia coli SURE cells (Stratagene). The ligation regenerates the NotI site, but not the MspI site, leaving CGG as the first 3 bases of the 5' end of all PCR products obtained. Each library contained in excess of 5 x 10 5 recombinants to ensure a high likelihood that the 3' ends of all mRNAs with concentrations of 0.001% or greater were multiply represented. Plasmid 10 preps (Qiagen) were made from the cDNA library of each sample under study. An aliquot of each library was digested with MspI, which effects linearization by cleavage at several sites within the parent vector while leaving the 3' cDNA inserts and their flanking sequences, including the T3 promoter, intact. The product was incubated 15 with T3 RNA polymerase (MEGAscript kit, Ambion) to generate antisense cRNA transcripts of the cloned inserts containing known vector sequences abutting the MspI and NotI sites from the original cDNAs. At this stage, each of the cRNA preparations was processed in a three-step 20 fashion. In step one, 250ng of cRNA was converted to first-strand cDNA using the 5' RT primer (A-G-G-T-C-G-A-C-G-G-T-A-T-C-G-G, (SEQ ID NO: 21). In step two, 400 pg of cDNA product was used as PCR template in four separate reactions with each of the four 5' PCR primers of the form G-G-T-C-G-A-C-G-G-T-A-T-C-G-G-N (SEQ ID NO: 22), each paired with a "universal" 3' PCR primer G-A-G-C-T-C-C-A-C-C-G-C-G-G-T 25 (SEQ ID NO: 23) to yield four sets of PCR reaction products ("NI reaction products"). In step three, the product of each subpool was further divided into 64 subsubpools (2ng in 20gl) for the second PCR reaction. This PCR reaction comprised adding 100 ng of the fluoresceinated "universal" 3' PCR primer (SEQ ID NO: 23) conjugated to 6-FAM 30 and 100 ng of the appropriate 5' PCR primer of the form C-G-A-C-G-G-T-A-T-C-G-G N-N-N-N (SEQ ID NO: 24), and using a program that included an annealing step at a 82 WO 01/30972 PCT/USOO/29690 temperature X slightly above the Tm of each 5' PCR primer to minimize artifactual mispriming and promote high fidelity copying. Each polymerase chain reaction step was performed in the presence of TaqStart antibody (Clonetech). 5 The products ("N4 reaction products") from the final polymerase chain reaction step for each of the tissue samples were resolved on a series of denaturing DNA sequencing gels using the automated ABI Prizm 377 sequencer. Data were collected using the GeneScan software package (ABI) and normalized for amplitude and migration. Complete execution of this series of reactions generated 64 product subpools for each of 10 the four pools established by the 5' PCR primers of the first PCR reaction, for a total of 256 product subpools for the entire 5' PCR primer set of the second PCR reaction. The mRNA samples from each timepoint as described above were analyzed. Table 1 is a summary of the expression levels of 495 mRNAs determined from cDNA. 15 These cDNA molecules are identified by their digital address, that is, a partial 5' terminus nucleotide sequence coupled with the length of the molecule, as well as the relative amount of the molecule produced at different time intervals after treatment. The 5' terminus partial nucleotide sequence is determined by the recognition site for MspI (CC GG) and the nucleotide sequence of the parsing bases of the 5' PCR primer used in the 20 final PCR step. The digital address length of the fragment was determined by interpolation on a standard curve and, as such, may vary ± 1-2 b.p. from the actual length as determined by sequencing. For example, the entry in Table 1 that describes a DNA molecule identified by the 25 digital address MspI AGTA, is further characterized as having a 5' terminus partial nucleotide sequence of CGGAGTA and a digital address length of 106 b.p. The DNA molecule identified as MspI AGTA 106 is further described as being expressed at increasing levels after both acute and chronic treatment with clozapine (see Fig. 1). Additionally, the DNA molecule identified as MspI AGTA 106 is described by its 30 nucleotide sequence, which coresponds with SEQ ID NO: 1. 83 WO 01/30972 PCT/USOO/29690 Similarly, the other DNA molecules identified in Table 1 by their MspI digital addresses are further characterized by: 1) the level of gene expression in the striatum/nucleus accumbens of mice without clozapine treatment (control), 2) the level of gene expression in the striatum/nucleus accumbens of mice treated with clozapine for 45 5 minutes, 3) the level of gene expression in the striatum/nucleus accumbens of mice treated with clozapine for 7 hours, 4) the level of gene expression in the striatum/nucleus accumbens of mice treated with clozapine for 5 days, 5) the level of gene expression in the striatum/nucleus accumbens of mice treated with clozapine for 12 days, 6) the level of gene expression in the striatum/nucleus accumbens of mice treated with clozapine for 10 14 days. Some products, which were also differentially represented, appeared to migrate in positions that suggest that the products were novel based on comparison to data extracted from GenBank. The sequences of such products were determined by one of two 15 methods: cloning or direct sequencing of the PCR products. Additionally, several of the isolated clones were further characterized as shown in Table 2 and their nucleotide sequences are provided as SEQ ID NOs: 1-19; 49-52; 57-72 and 107 in the Sequence Listing below. 20 The sequences of SEQ ID NOs: 1-19; 49-52; 57-72 and 107 have had the MsPI site found in the native state of the corresponding RNA indicated by the addition of a "C" to the 5' of the sequence. As noted above, the ligation of the sequence into a vector does not regenerate the MspI site; the experimentally determined sequence reported herein has 25 C-G-G as the first bases of the 5' end. The data shown in Figure 1 were generated with a 5'-PCR primer (C-G-A-C-G G-T-A-T-C-G-G-A-G-T-A; SEQ ID NO: 94) paired with the "universal" 3' primer (SEQ ID NO: 23) labeled with 6-carboxyfluorescein (6FAM, ABI) at the 5' terminus. PCR 30 reaction products were resolved by gel electrophoresis on 4.5% acrylamide gels and fluorescence data acquired on AB1377 automated sequencers. Data were analyzed using 84 WO 01/30972 PCT/USOO/29690 GeneScan software (Perkin-Elmer). The sequences of the PCR products were determined using standard techniques. The results of TOGA analysis using a 5' PCR primer with parsing bases AGTA 5 (SEQ ID NO.: 94) are shown in Figure 1, which shows the PCR products produced from mRNA isolated from the striatum/nucleus accumbens of mice treated with clozapine for various lengths of time as described above. The vertical index line indicates a PCR product of about 106 b.p. that is present in control cells, and whose expression increases when the striatum/nucleus accumbens of mice are treated with clozapine for 45 minutes, 10 7 hours, 5 days, 12 days, and 14 days. Cloning DSTs Without A Candidate Match and Verification of the Cloned DSTs Using Extended TOGA Method 15 In suitable cases, the PCR product was isolated, cloned into a TOPO vector (Invitrogen) and sequenced on both strands. The database matches for each cloned DST sequence are listed in Table 2. In order to verify that the cloned product corresponds to the TOGA peak of interest, the extended TOGA assay was performed for each DST. PCR primers were designed based on the determined sequences and PCR was performed 20 using the NI PCR reaction products as a substrate. Oligonucleotides were synthesized with the sequence G-A-T-C-G-A-A-T-C extended at the 3' end with a partial MspI site (C-G-G), and an additional 18 adjacent nucleotides from the determined sequence of the cloned PCR product or DST. For example, for the PCR product with the digital address MspI AGTA 106 (SEQ ID NO: 1), the 5' PCR primer was G-A-T-C-G-A-A-T-C-C-G-G 25 A-G-T-A-C-A-G-T-G-A-C-T-T-T-G-A-G-T (SEQ ID NO: 28). This 5' PCR primer was paired with the fluorescent labeled universal 3' PCR primer (SEQ ID NO: 23) in a PCR reaction using the PCR NI reaction product as substrate. The length of the PCR product generated with the clone specific primer (SEQ ID 30 NO: 28) was compared to the length of the original PCR product that was produced in the TOGA reaction as shown in Figure 2. For CLZ_3 (SEQ ID NO: 1), the upper panel 85 WO 01/30972 PCT/USOO/29690 (Figure 2A) shows the PCR product generated with the clone specific primer (SEQ ID NO: 28) and the fluorescent labeled universal 3' PCR primer (SEQ ID NO: 23). Figure 2B shows the PCR products produced in the original TOGA reaction using a 5' PCR primer C-G-A-C-G-G-T-A-T-C-G-G-A-G-T-A (SEQ ID NO: 94), and the fluorescent 5 labeled universal 3' PCR primer. In the bottom panel (Figure 2C), the traces from the top and middle panels are overlaid, demonstrating that the PCR product produced using an extended primer based on the cloned sequence is the same length as the original PCR product. Other DST clones verified using this method include cases (CLZ_5, SEQ ID NO: 2; CLZ_8, SEQ ID NO: 3; CLZ_10, SEQ ID NO: 4; CLZ_12, SEQ ID NO: 5; 10 CLZ_15, SEQ ID NO: 6; CLZ_24, SEQ ID NO: 7; CLZ_33, SEQ ID NO: 8; CLZ_34, SEQ ID NO: 9; CLZ_37, SEQ ID NO: 10; CLZ_38, SEQ ID NO: 11; CLZ_40, SEQ ID NO: 12, CLZ_6, SEQ ID NO: 14; CLZ_16, SEQ ID NO: 15; CLZ_22, SEQ ID NO: 16; CLZ_32, SEQ ID NO: 17; CLZ_36, SEQ ID NO: 18; CLZ_42, SEQ ID NO: 19; CLZ_18, SEQ ID NO: 57; CLZ_43, SEQ ID NO: 58; CLZ_44, SEQ ID NO: 59; 15 CLZ_47, SEQ ID NO: 60; CLZ_48, SEQ ID NO: 61; CLZ 49, SEQ ID NO: 62; CLZ_50, SEQ ID NO: 63; CLZ_51, SEQ ID NO: 64; CLZ_52, SEQ ID NO: 65; CLZ_56, SEQ ID NO: 67; CLZ_57, SEQ ID NO: 68; CLZ_60, SEQ ID NO: 69, and CLZ_64, SEQ ID NO: 70). Table 3 contains primers generated from each of the cloned DSTs used in such studies. 20 Direct Sequencing of TOGA Generated PCR products and Verification by Extended TOGA Method In other cases, the TOGA PCR product was sequenced using a modification of a 25 direct sequencing methodology (Innis et al., Proc. Nat'l. Acad. Sci., 85: 9436-9440 (1988)). PCR products corresponding to DSTs were gel purified and PCR amplified again to incorporate sequencing primers at 5' and 3' ends. The sequence addition was 30 accomplished through 5' and 3' ds-primers containing M13 sequencing primer sequences (M13 forward and M13 reverse respectively) at their 5' ends, followed by a linker 86 WO 01/30972 PCT/USOO/29690 sequence and a sequence complementary to the DST ends. Using the Clontech Taq Start antibody system, a master mix containing all components except the gel purified PCR product template was prepared, which contained sterile H 2 0, I OX PCR II buffer, 1 OmM dNTP, 25 mM MgC1 2 , AmpliTaq/Antibody mix (1.1 pg/pl Taq antibody, 5 U/pl 5 AmpliTaq), 100 ng/ptl of 5' ds-primer (5' TCC CAG TCA CGA CGT TGT AAA ACG ACG GCT CAT ATG AAT TAG GTG ACC GAC GGT ATC GG 3', SEQ ID NO: 89), and 100 ng/pl of 3' ds-primer (5' CAG CGG ATA ACA ATT TCA CAC AGG GAG CTC CAC CGC GGT GGC GGC C 3', SEQ ID NO: 90). After addition of the PCR template, PCR was performed using the following program: 94"C, 4 minutes and 25 10 cycles of 94'C, 20 seconds; 65 0 C, 20 seconds; 72 0 C, 20 seconds; and 72 0 C 4 minutes. The resulting amplified adapted PCR product was gel purified as described above. The purified ds-extended PCR product was sequenced using a standard protocol for ABI 3700 sequencing. Briefly, triplicate reactions in forward and reverse orientation 15 (6 total reactions) were prepared, each reaction containing 5 d of gel purified ds extended N5 PCR product as template. In addition, the sequencing reactions contained 2 ptl 2.5X sequencing buffer, 2 pl Big Dye Terminator mix, 1 pl of either the 5' sequencing primer (5' CCC AGT CAC GAC GTT GTA AAA CG 3', SEQ ID NO: 91), or the 3' sequencing primer (5' TTT TTT TTT TTT TTT TTT V 3', where V=A, C, or G, SEQ ID 20 NO: 92) in a total volume of 10 pl. In an alternate embodiment, the 3' sequencing primer was the sequence 5' GGT GGC GGC CGC AGG AAT TTT TTT TTT TTT TTT TT 3', (SEQ ID NO: 93). PCR was performed using the following thermal cycling program: 96"C, 2 minutes and 29 25 cycles of 96'C, 15 seconds; 50"C, 15 seconds; 60 0 C, 4 minutes. The sequences for (CLZ_62, SEQ ID NO: 71 and CLZ_65, SEQ ID NO: 72) were determined by this method. Table 2 contains the database matches for the sequences determined by this method. 30 87 WO 01/30972 PCT/USOO/29690 In order to verify that the sequences determined by direct sequencing derive from the PCR product of interest, PCR primers were designed based on the sequences determined by direct sequencing, and PCR reactions were performed using the N1 TOGA PCR reaction products as substrate, as described above for the sequences cloned into the 5 TOPO vector. In short, oligonucleotides were synthesized with the sequence G-A-T-C G-A-A-T-C extended at the 3' end with a partial MspI site (C-G-G) and an additional 18 nucleotides adjacent to the partial MspI site from the sequence determined by direct sequencing. The 5' PCR primers were paired with the fluorescent labeled universal 3' PCR primer (SEQ ID NO: 23) in PCR reactions with the N1 TOGA PCR reaction 10 product as template. The lengths of these PCR products were compared to the length of the PCR products of interest. Table 3 contains the sequences of the primers used in these studies. Verification of a Candidate Match Using Extended TOGA Method 15 In four cases, CLZ_17, (SEQ ID NO: 49); CLZ_26, (SEQ ID NO: 50); CLZ_28, (SEQ ID NO: 51); and CLZ_ 58 (SEQ ID NO: 52) the sequences listed for the TOGA PCR products were derived from candidate matches to sequences present in available Genbank, EST, or proprietary databases. Table 4 lists the candidate matches for each by 20 accession number of the Genbank entry or by the accession numbers of a set of computer-assembled ESTs used to create a consensus sequence. To determine whether the TOGA PCR products of interest were derived from the sequence of the candidate match, PCR primers were designed with the sequence G-A-T 25 C-G-A-A-T-C extended at the 3' end with a partial MspI site (C-G-G), and an additional 18 nucleotides adjacent to the terminal MspI site in the candidate match sequence. Each extended primer is combined with the fluorescent labeled universal 3' PCR primer (SEQ ID NO: 23) in a PCR reaction with the product of the first TOGA PCR reaction (NI reaction products) as the template. The PCR products obtained using an extended primer 30 and the universal 3' primer were compared to products obtained using the original TOGA PCR primers. Primers designed for such studies are shown in Table 4 along with the accession numbers of sequences used to derive the primer sequences. 88 WO 01/30972 PCT/USOO/29690 EXAMPLE 2 Characterization of CLZ 5 (apoD) 5 Another example of TOGA analysis is shown in Figure 3. In Figure 3, a peak at about 201 is indicated, identified by digital address MspI CACC 201 when a 5' PCR primer (SEQ ID NO: 25) was paired with SEQ ID NO: 23 to produce the panel of PCR products. The PCR product was cloned and sequenced as described in Example 1. To verify the identity of the isolated clone (SEQ ID NO: 2), oligonucleotides were 10 synthesized corresponding to the 5' PCR primer in the second PCR step for each candidate extended at the 3' end with an additional 12-15 nucleotides from the cloned sequence. In this case the 5' PCR primer was G-A-T-C-G-A-A-T-C-C-G-G-C-A-C-C-T A-C-T-G-G-A-T-C-C-T-G-G (SEQ ID NO: 29). This 5' PCR primer were paired with the fluorescently labeled 3' PCR primer (SEQ ID NO: 23) in PCRs using the cDNA 15 produced in the first PCR reaction as substrate. As shown in Tables 2 and 3, the CLZ_5 clone (CACC 201) described above corresponds with GenBank sequence X82648, which is identified as a mouse apolipoprotein D (apoD) sequence. Other corresponding apoD GenBank sequences 20 include L39123 (mouse), X55572 (rat), NM_001647 (human). Northern Blot analyses were performed to determine the effect of clozapine and haloperidol on apoD expression in mouse striatum/nucleus accumbens. Also, in situ hybridization analyses were performed to determine the pattern of apoD expression in control and clozapine-treated mouse striatum/nucleus accumbens. 25 Male C57B/6J mice (20-28 g) were housed in groups of four on a standard 12/12 hour light-dark cycle with ad libitum access to standard laboratory chow and tap water. The same experimental paradigm used in Example 1 was used for the Northern Blot analyses. Briefly, the control group mice received a single injection of sterile saline (0.1 30 ml volume), or no injection, and were sacrificed after 45 minutes. The mice subjected to acute neuroleptic treatment were given a single intraperitoneal injection of the typical neuroleptic, haloperidol, (4 mg/kg) or the atypical neuroleptic, clozapine (7.5 mg/kg) and 89 WO 01/30972 PCT/USOO/29690 sacrificed after 45 minutes or 7 hours, as described in Example 1. The mice subjected to chronic neuroleptic treatment received daily subcutaneous injections of haloperidol (4 mg/kg) for 10 days or 14 days, or received daily injections of clozapine (7.5 mg/kg) for 5 days, 12 days or 14 days. All animals were sacrificed in their cages with CO 2 at the 5 indicated times. Brains were rapidly removed and placed on ice. The striatum, including the nucleus accumbens, were dissected out and placed in ice-cold phosphate-buffered saline. The cytoplasmic RNA was isolated by phenol:chloroform extraction of the homogenized tissue according to the method described in Schibler et al., J. MoL. Bio., 142, 93-116 (1980). Poly A enriched mRNA was prepared from cytoplasmic RNA using 10 well-known methods of oligo dT chromatography. Shown in Fig. 4, Northern Blot analysis was performed using 2 ptg poly A enriched mRNA extracted from the striatum/nucleus accumbens of control mice and clozapine-treated mice. The mRNA transcripts were fractionated by electrophoresis on a 15 1.5% agarose gel containing formaldehyde, transferred to a biotrans membrane by the method of Thomas (Thomas, P. S., Proc. NatL. Acad. Sci., 77,5201-5215 (1980)), and prehybridized for 30 minutes in Expresshyb (Clonetech). A 160 bp insert of CLZ_5 (25 100 ng) was labeled with [a- 32 P]-d CTP by oligonucleotide labeling to specific activities of approximately 5x10 8 cpm/pg, added to the prehybridization solution and incubated for 20 1 hour. Filters were washed to high stringency (0.2 X SSC) (1 X SSC: 0.015 M NaCl and 0.00 15 M Na citrate) at 68*C then exposed to Kodak X-AR film (Eastman Kodak, Rochester, NY) for up to 1 week. Densitrometry analysis on Northern blots was performed by ImageQuant software. 25 As can be seen in Fig. 4, a 900 bp mRNA was detected in control and clozapine treated mice which corresponds with the apoD gene. The apoD mRNA expression is progressively up-regulated with clozapine treatment over the two-week time course. It is possible that clozapine may mediate its antipsychotic effect through the regulation of apoD. Alternatively, apoD may be co-regulated by clozapine, in parallel with the 30 mechanism of the clozapine therapeutic effects, and can serve as an indicator of clozapine bioactive levels. 90 WO 01/30972 PCT/USOO/29690 Shown in Fig. 5, Northern Blot analysis was performed using mRNA extracted from the striatun/nucleus accumbens of control mice and haloperidol-treated mice using the above-described method and the same 32 P-radiolabelled probe. A 900 bp mRNA was 5 detected in control and haloperidol-treated mice which corresponds with the apoD gene. Interestingly, apoD mRNA expression is slightly down-regulated with acute and chronic haloperidol treatment. These results reveal that clozapine and haloperidol have a differential effect on apoD expression. 10 Figure 6 is a graphical representation comparing the results of the clozapine treatment TOGA analysis of clone CLZ_5 (CACC 201) shown in Fig. 4 and the clozapine treatment Northern Blot analysis of clone CLZ_5 shown in Figure 5. The Northern Blot was imaged using a phosphoimager to determine the amount of apoD mRNA in each clozapine-treated sample relative to the amount of mRNA in the control 15 sample. As can be seen, the clozapine treatment TOGA analysis shows correlation with the clozapine treatment Northern Blot analysis. Figure 7A-C shows an in situ hybridization analysis, demonstrating the apoD expression in mouse brain. The in situ hybridization was performed on free-floating 20 sections (25 piM thick) as described (Thomas et al., J Neurosci. Res., 52, 118-124 (1998)). Coronal sections were hybridized at 55 0 C for 16 hours with an 5 S-labeled, single-stranded 160 bp antisense cRNA probe of CLZ_5 at 107 cpm/ml. The probe was synthesized from the 3'-ended cDNA TOGA clone CLZ-5 using the Maxiscript Transcription Kit (Ambion, Austin, TX). Excess probe was removed by washing with 2 25 X SSC (I X SSC = 0.015 M NaCl/0.0015 M Na citrate) containing 14 mM s mercaptoethanol (30 minutes), followed by incubation with 4 ptg/ml ribonuclease in 0.5 M NaC1/0.05 M EDTA/0.05 M Tris-HC1, pH 7.5, for 1 hour at 37*C. High stringency washes were carried out at 55*C for 2 hours in 0.5 X SSC/50% formamide/0.01 M p mercaptoethanol, and then at 68 0 C for 1 hour in 0.1 X SSC/0.01 M P 30 mercaptoethanol/0.5% sarkosyl. Slices were mounted onto gelatin-coated slides and dehydrated with ethanol and chloroform before autoradiography. Slides were exposed 91 WO 01/30972 PCT/USOO/29690 for 1-4 days on Kodak X-AR film and then dipped in Ilford K-5 emulsion. After 4 weeks, slides were developed with Kodak D19 developer, fixed, and counterstained with Richardson's blue stain. 5 Fig. 7A shows CLZ-5 (apoD) mRNA expression in mouse anterior brain, 7B shows apoD mRNA expression in midbrain and 7C shows apoD expression in posterior brain. In all brain sections apoD is expressed by astroglial cells, pial cells, perivascular fibroblasts and scattered neurons. This is consistent with previous studies examining the expression of apoD in mice, rabbits and humans (Yoshida et al., DNA and Cell Biology, 10 15, 873-882 (1996); Provost et al., J. Lipid Res., 32, 1959-1970 (1991); Navarro et al., Neurosci. Lett., 254, 17-20 (1998). The Northern blot results (Figures 4 and 6) indicated that apoD was induced by clozapine in the striatum of mouse brain. To investigate additional sites of apoD 15 induction, in situ hybridization analysis was performed on brains from saline- and clozapine-treated mice. Figure 8A-I presents an in situ hybridization analysis, showing clone CLZ_5 (apoD) mRNA expression in mouse anterior (8A-C), mid (8D-F), and posterior (8G-I) brain following saline treatment (top row) or clozapine treatment (7.5 mg/kg) for 5 days (middle row) and 14 days (bottom row), using previously described 20 methods. Animals were sacrificed by intracardial perfusion with 4% paraformaldehyde and the brains removed, post-fixed for 12 hours, cryoprotected with 30% sucrose and rapidly frozen at -70C. At low magnification, increases in apoD mRNA were observed at both five days and two weeks of clozapine treatment in the striatum, cortex, globus pallidus (GP), and thalamus. Increases in apoD expression were also detected in white 25 matter tracts, predominantly the corpus callosum (cc), anterior commissure, internal capsule (ic) and optic tract (opt). At high magnification, it was evident that the increased apoD hybridization signal in the striatum, globus pallidus, and thalamus of the drug treated animals was primarily due to an increase in the number of cells expressing detectable apoD, although some cells with higher apoD expression were also observed. 30 92 WO 01/30972 PCT/USOO/29690 Using a monoclonal antibody directed against full-length apoD, immunohistochemistry analyses were performed to evaluate changes in apoD protein expression in response to clozapine. Increase in protein expression correlated well with increases in mRNA expression (data not shown). Combined in situ hybridization and 5 immunohistochemical studies demonstrated that increases in apoD levels were localized primarily to neurons and astrocytes of the striatum and oligodendrocytes in various white matter tracts throughout the brain. Figure 9A-H shows a darkfield photomicrograph demonstrating upregulated apoD 10 mRNA expression in various brain regions, including the corpus callosum (cc, Fig. 9A, E); caudate putamen (CPu, Fig. 9B, 7F); anterior commissure (aca, Fig. 9C, 9G); and globus pallidus (GP, Fig. 9D, 911). In situ hybridizations were performed as described above, using an antisense 35 S-labeled apoD riboprobe on brains from control (Fig. 9A-D) and clozapine-treated (Fig. 9E-H) animals. The observed upregulation of apoD was due 15 to an increase in the amount of apoD expressed per cell. Figure 1 OA, B shows a darkfield photomicrograph demonstrating upregulated apoD mRNA expression in the internal capsule (ic). Figure 10C, D shows a brightfield view of the optic tract (opt) demonstrating upregulation of apoD expression in 20 oligodendrocytes. In situ hybridizations were performed as described above, using an antisense 35 S-labeled apoD riboprobe on brains from control (10A, C) and clozapine treated (10B, D) animals. As shown in Fig. 1OD, the cells prominantly expressing apoD in the optic tract have a box-like morphology and are lined up in a serial array, presumably along axonal tracts. Such features are characteristic of oligodendrocytes, 25 which synthesize the insulating myelin coating of nerve fibers. In situ hybridization experiments performed on brains from haloperidol-treated mice did not reveal substantial increases in apoD expression in gray or white matter regions (data not shown). White matter tracts comprise nerve fiber bundles connecting different regions of 30 the brain. The predominant cells in these regions are astrocytes and oligodendrocytes, both of which have been shown to express apoD (Boyles et al., JLipid Res 31:2243-2256 93 WO 01/30972 PCT/USOO/29690 (1990); Navarro et al., Neurosci Lett 254:17-20 (1995); Provost et al., JLipid Res 32 (1991)). To determine which cell types are responsible for the increase in apoD signal, co-localization studies were performed using a 35 S-labeled apoD riboprobe in combination with either an antibody specific for an astrocyte marker, glial fibrillary 5 acidic protein (GFAP), or an antibody specific for an oligodendrocyte marker, 2', 3' cyclic nucleotide 3'-phosphodiesterase (CNP) (Boehringer Mannheim, Germany). The immunoreaction was detected with Vectastain ABC TM kit (Vector Laboratory, Inc., Burlingame, CA) according to the manufacturer's instructions. 10 Free floating brain sections were incubated with blocking solution (4% bovine serum albumin in 0.1% Triton X-100/PBS) for 2 hours at room temperature, followed by incubation with anti-GFAP or anti-CNP antiserum (dilution 1:500) in blocking solution for 16-20 hours at 4"C. Sections were then washed with 0.1% Triton X-100/PBS and incubated with secondary biotinylated antibody (1:200 dilution in blocking solution) for 2 15 hours at room temperature. The sections were then washed with 0.1% Triton X-100/PBS, incubated for 1 hour with ABC reagent (1:1 in blocking solution) and finally washed with 0.1% Triton X-100/PBS. Enzymatic development was performed in 0.05% diaminobenzene in PBS containing 0.003% hydrogen peroxide for 3-5 minutes. 20 Fig. 11 shows sections of striatum and optic tract in control and clozapine-treated animals. Thick arrows indicate the co-localization of GFAP and apoD, while thin arrows indicate the expression of apoD alone. Fig. 11 A, B shows that in untreated striatum, many GFAP-positive cells in both gray and white matter regions are positive for apoD. Fig. lI D, E shows that in brain from clozapine-treated animals, an increase in the amount 25 of apoD was observed in a small subset of GFAP-positive cells in the striatum. Additionally, there was an increase in the number of non-GFAP-positive cells expressing apoD in the striatum, as well as the globus pallidus and thalamus which are presumptively neurons, based on size and morphology. 30 Fig. 11 C, F shows GFAP and apoD co-localization in the optic tract in control (11 C) and clozapine-treated (1 IF) animals. While some astrocytes express apoD mRNA, 94 WO 01/30972 PCT/USOO/29690 the cells responsible for the predominant apoD transcript upregulation did not label with GFAP and thus are likely oligodendrocytes. In other white matter regions, such as the corpus callosum, anterior commissure and internal capsule, the non-GFAP expressing cells that express apoD are likely to be oligodendrocytes as well, although expression in 5 microglia can not be ruled out. Fig. 11 G, H shows apoD immunohistochemistry with an anti-human apoD primary antibody (Novocastra, Newcastle, UK) in the optic tract of control saline (11 G) and clozapine-treated animals (1 1H). Co-localization studies performed using anti-CNP antibody showed CNP 10 immunoreactivity in white matter tracts throughout the CNS which correlated with areas of apoD mRNA hybridization signals, indicating the expression of apoD in oligodendrocytes. However, within the gray matter regions of the striatum, there was no co-localization consistent with the neuronal accumulation of apoD (data not shown). 15 Figure 12 shows a Northern Blot analysis of clone CLZ_5 expression in cultured glial cells treated with clozapine (100 nM and 1 ptM) for 1 day or 7 days. Glial cell cultures were produced from postnatal (day 2) rats. The cells were treated with different concentrations of clozapine for different lengths of time before mRNA extraction as follows: A= control (no clozapine), B= 100 nM clozapine, 1 day, C= 1pM clozapine, 1 20 day, D= 100 nM clozapine, 1 week, E= IiM clozapine, 1 week. 20 ig of total cytoplasmic RNA from glial cell cultures were electrrophoresed on a 1.5% agarose gel containing formaldehyde, blotted, and probed as previously described. Interestingly, apoD mRNA levels were down-regulated in mixed glial cell cultures treated with clozapine (both 100 nM and 1 pM) for 1 week, suggesting that perhaps neurons and glia 25 display different mechanisms for apoD regulation. TOGA methodology, Northern blot analyses, and in situ hybridization studies have demonstrated an increase in apoD mRNA expression in both white and gray matter regions of mouse brain in response to chronic clozapine administration. Colocalization 30 studies, combining in situ hybridization and imunohistochemistry methods have revealed that apoD mRNA levels are increased in both neurons and glial cells with clozapine 95 WO 01/30972 PCT/USOO/29690 administration. The evidence indicates that the glial cells responsible for the most dramatic increases in apoD expression are primarily oligodendrocytes, but a subset of astrocytes also have increased apoD expression after clozapine treatment. In contrast, TOGA, Northern blot and in situ hybridization analyses showed that apoD expression 5 was not affected by haloperidol treatment. In addition to the mouse studies described above which show that apoD is regulated by chronic antipsychotic drug administration, studies using schizophrenic and bipolar human subjects showed that apoD expression is increased in the prefrontal cortex 10 of such patients. The combined results suggest that apoD is a marker for neuropathology associated with psychiatric disorders and therefore can be used to target abnormalities in specific anatomical brain regions. ApoD was initially identified as a constituent of plasma high-density lipoproteins 15 (HDLs), which also contain phospholipids, cholesterol and fatty acids (McConathy et al., Fed. Eur. Biochem. Soc. Lett., 37: 178 (1973)). In the blood, apoD is thought to play a role in reverse cholesterol transport, the removal of excess cholesterol from tissues to the liver for catabolism (Oram et al., J. Lipid. Res., 37: (1996)). In addition to abundant expression in human serum, apoD is major protein component in cyst fluid from women 20 with human breast cystic disease (Balbin et al., Biochem. J., 271: 803 (1990)) and also is widely expressed in numerous tissues, including liver, kidney, intestine, spleen and brain (Drayna et al., J. Bio. Chem., 261: (1986)). In the CNS of humans, as in other species (Provost et al., J. Lipid Res., 32: (1991); Seguin et al., Mol. Brain Res., 30: 242 (1995); Smith et al., J Lipid Res., 31: 995 (1990)), apoD is expressed primarily in glial cells, pial 25 cells, perivascular cells, and some neuronal populations (Navarro et al., Neurosci. Lett., 254: 17 (1995); Kalman et al., Neurol. Res., 22: 330 (2000)). The physiological role for apoD within the CNS is not known, however, it has been shown to bind several hydrophobic ligands, including sterols and steroid hormones (Dilley et al., Breast Canc. Res. Treat., 16: 253 (1990); Lea, 0. A., Steroids, 52: 337 (1988)) suggesting a role in 30 extracellular lipid transport in the brain. ApoD has also been shown to bind arachidonic acid Morais-Cabral et al., FEBS Lett., 366: 53 (1995)) implicating it in functions 96 WO 01/30972 PCT/USOO/29690 associated with cell membrane remodeling and prostaglandin synthesis. In the regenerating sciatic nerve, a process that involves massive membrane degradation and lipid release, apoD concentrations are increased 500-fold (Boyles et al., J. Bio. Chem., 265: 17805 (1990)). Recent reports have also demonstrated an increase in apoD 5 expression in rat brain after experimental and chemical lesioning of the entorhinal cortex and hippocampus, respectively (Ong et al., Neurosci., 79:359 (1997); Terisse et al., Mo. Brain Res., 70: 26 (1999)). Additionally, in humans, apoD accumulates in the cerebrospinal fluid and hippocampi of patients with Alzheimer's, and other neurological diseases (Terisse et al., J. Neurochem., 71: 1643 (1998)). Hence, apoD may be 10 functioning during pathological situations or its expression may represent an effort to compensate for neuropathology associated with such insults. The pattern of apoD expression in the brain suggests that apoD may play an important role in psychotic disease. It is widely believed that imbalances in basal ganglia 15 circuitry contribute to psychotic behaviors and that blockade of specific receptors in these regions is responsible for neuroleptic action. The neuronal increases in apoD mRNA expression observed in neurons of the striatum and globus pallidus are consistent with this hypothesis. 20 In addition, the apoD induction observed in the internal capsule is of particular interest. The internal capsule consists of massive nerve fibers connecting the thalamus to the cortex and is an area of convergence for the fiber tracts running transversely through the striatum. The thalamus is a relay station for virtually all information passing to the cortex and coordinated cortico-thalamic activity is essential for normal consciousness. 25 Recent theories have associated psychotic behavior with disruptions in cortico-thalamic oscillations. An upregulation of apoD expression in the internal capsule may play a role in restoring the proper balance of neuronal communication. In addition, abnormal lipid neurochemistry resulting from abnormal lipid 30 transport or metabolism has been associated with psychotic disease, such as schizophrenia (Walker et al., Br. J. Psych., 174, 101-104 (1999)). Relating impaired 97 WO 01/30972 PCT/USOO/29690 cholesterol metabolism with psychotic disease, a number of reports have described psychoses as an initial manifestation of Neimann-Pick Disease, type C (Campo, et al., Develop. Med. and Child Neurol., 40, 126-129 (1998); Shulman, et al., Neurology, 45, 1739-1743 (1995); Turpin, et al., Dev. Neurosci., 13, 304-306 (1991)), which is an 5 autosomal recessive disease associated with abnormal cholesterol metabolism (Yoshida et al., DNA and Cell Biology, 15, 873-882 (1996)). Further reports have suggested that myelin dysfunction may cause mental illness. Given that the majority of cholesterol in the brain is incorporated into myelin, abnormal cholesterol metabolism may result in myelin dysfunction. Myelin acts as an insulator along nerve axons allowing for the rapid 10 propagation of action potentials along nerve fibers. Molecular abnormalities of myelin may result in the dysregulated neural connectivity that has been hypothesized to be causative in mental illnesses (Weickert, et al., Schizophrenia Bull., 24, 303-316 (1998)). While the physiological function of apoD in the CNS is not clear, several lines of 15 evidence suggest a role for apoD as a vehicle for extracellular lipid transport and lipid movement, particularly cholesterol, in the nervous system. ApoD is a constituent of plasma high-density lipoproteins (HDLs), which also contain phospholipids, cholesterol and fatty acids. While not much is known about HDL compared to the other plasma lipoproteins, LDL and VLDL, it is widely believed that HDLs protect against 20 cardiovascular disease by removing excess cholesterol from cells of arterial walls. This removal involves the direct interaction of HDL lipoproteins with plasma membrane domains and subsequent transport to the liver for catabolism (Oram, et al., J. Lipid Res., 37, 2473-2491 (1996)). Additionally, apoD is synthesized and secreted by cultured astrocytes, which secretion has been shown to increase in the presence of cholesterol 25 derivatives (Patel, et al., Neuroreport 6, 653-657 (1995)). Further, it has also been demonstrated that apoD levels are increased in Niemann Pick Disease, type C, which is associated with elevated levels of cholesterol. These studies provide evidence of a functionally significant role for apoD in cholesterol transport in the CNS. 30 In addition to the studies correlating cholesterol levels and psychotic behavior, other studies have found a correlation between cholesterol levels and treatment with 98 WO 01/30972 PCT/USOO/29690 neuroleptics. For example, reports dating back to 1960 have demonstrated an increase in the serum cholesterol of patients treated with conventional neuroleptics, such as chlorpromazine and haloperidol (Spivak et al., Clin. Neuropharm., 22, 98-101 (1999). Fleischhacker et al., Pharmacopsychiatry, 19, 111-114 (1986); Clark et al., Clin. Pharm. 5 and Therapeutics, 11, 883-889 (1970)). However similar increases are not observed with the newer, atypical antipsychotics, such as fluperlapine and clozapine (Spivak et al., Clin. Neuropharm., 22, 98-101 (1999). Fleischhacker et al., Pharmacopsychiatry, 19, 111-114 (1986); Boston, et al., Biol. Psych., 40, 542-543 (1996)). Interestingly, the present results reveal that clozapine and haloperidol have a differential effect on apoD expression, which 10 may account for the observed differences in cholesterol regulation. While the mechanism for these cholesterol changes is not known, the present data suggest that neuroleptic induced changes in apoD expression combined with the ability of apoD to bind cholesterol may provide an explanation for the neuroleptic-induced changes in cholesterol levels. 15 In addition to studies relating to cholesterol movement, reports have focused on the link between disrupted phospholipid and fatty acid metabolism and psychiatric disorders (for a review see Horrobin, et al., Prostaglandins, Leukotrienes and Essential Fatty Acids, 60, 141-167 (1999)). For example, numerous studies have reported 20 differences in levels of total membrane phospholipid content, fatty acid levels, cholesterol levels and cholesteryl esters in fibroblasts and/or frontal cortex of schizophrenics (Keshavan et al., JPsychiatry Res., 49, 89-95 (1993); Mahadik et al., Schizophrenia Res. 13, 239-247 (1994); Sengupta et al., Biochem. Med., 25, 267-275 (1981); Stevens, Schizophr. Bull., 6, 60-61 (1972)). Membrane phospholipids act as 25 precursors in numerous signaling systems (e.g., inositol phosphates, arachidonic acid, platelet activation factors, and eicosaniods) and comprise the membrane environment for neurotransmitter-mediated signal transduction. Thus, altered membrane phospholipid metabolism could have significant consequences for neuronal communication, resulting in behavioral abnormalities. 30 99 WO 01/30972 PCT/USOO/29690 Alterations in plasma membrane structure and function may result from the altered content and distribution of membrane lipids and fatty acids, such as arachidonic acid. Arachidonic acid is released by the action of numerous phospholipase enzymes, primarily phospholipase A2, and is a substrate for prostglandins and leukotriene 5 synthesis. While the molecular mechanisms underlying abnormalities in the complex system of phospolipid biochemistry are not known, several groups have demonstrated an increase in phospholipase A2 activity in the plasma and brains of schizophrenic patients (Gattaz et al., Biol. Psychiatry., 22, 421-426 (1987); Ross et al., Arch. Gen. Psychiatry., 54, 487-494 (1997); Ross et al., Brain Research, 821, 407-413 (1999)). In addition, 10 plasma phospholipase A2 levels have been shown to be decreased after neuroleptic therapy (Gattaz et al., Biol. Psychiatry, 22, 421-426 (1987)). Other molecular candidates implicated in psychotic disease include phospholipase C enzymes, diacyl glycerol kinases, and inositol phosphates (Horrobin et al., Prostaglandins, Leukotrienes and Essential Fatty Acids, 60, 141-167 (1999)). 15 Interestingly, in addition to binding cholesterol, apoD has been shown to specifically bind arachidonic acid. ApoD is an atypical apolipoprotein in that it does not share sequence homology with other apolipoproteins (Weech et al., Prog. Lipid Res., 30, 259-266 (1991)) but, rather, is a member of the lipocalin superfamily of proteins, which 20 function in the transport of small hydrophobic molecules, including sterols, steroid hormones, and arachidonic acid (Balbin et al., Biochem. J., 271, 803-807 (1990); Dilley et al., Breast Cancer Res. Treat., 16, 253-260 (1990); Lea, Steroids, 52, 337-338 (1988); Boyles et al., J. Lipid Res., 31, 2243-2256 (1990)). As a lipid binding protein, apoD can affect fatty acid composition, cholesterol levels and membrane phospholipids, all of 25 which will affect plasma membrane composition and structure. Also, since apoD specifically binds cholesterol, arachidonic acid and other lipids, alterations in the levels of apoD can affect lipid metabolism and signal transduction by affecting substrate availability for these pathways. 30 Further implicating the role of apoD in psychosis is the observation that apoD may have a chromosomal linkage with schizophrenia. The chromosomal location of 100 WO 01/30972 PCT/USOO/29690 apoD is 3q26. Genetic studies have implicated a potential association between schizophrenia and chromosome 3q, however the linkage is relatively inconsistent (reviewed by Maier, et al., Curr. Opin. Psych., 11, 19-25 (1998)). 5 Northern blot analysis on striata from haloperidol-treated mice did not reveal similar increases in apoD expression as clozapine. Schizophrenia is a heterogeneous disorder encompassing many subtypes. The observed differences in clinical efficacy between clozapine and haloperidol may reflect different subtypes of schizophrenia that are associated with different pathways or mechanisms. Thus, regulation of apoD may 10 represent a unique mechanism of action for clozapine. In this regard, a serotonin sub-type such as 5HT2a and 5HT 2 e may provide a pharmacological mechanism for clozapine's effect on apoD expression. Preliminary results demonstrate that treatment with ketanserin and mesulergine, 5HT 2 an 2 e and 5HT 2 c 15 selective antagonists respectively, results in an apparent upregulation of apoD mRNA expression in mouse brain. It is known that the striatum expresses a number of 5HT receptor subtypes, including the 5HT 2 c, which subtype may mediate clozapine's effect on apoD expression. In contrast, cultured glial cells or astrocytes do not appear to express 5HT 2 c receptors. Thus the downregulation observed in these cells may reflect actions at a 20 different 5HT subtype, such as 5HT 2 a, or a different receptor. Additionally, in hypertension studies, ketanserin has been associated with a decrease in total cholesterol levels and an upregulation of another apolipoprotein, apo Al (Loschiavo, et al., Int. J. Clin. Pharmacol. Ther. Toxicol., 28, 455-457 (1990)). The similar effects observed by both ketanserin and clozapine suggest that they may be working through the same 25 receptor subtype(s). The finding that apoD mRNA levels are increased by clozapine links apolipoproteins and the mechanism of action of neuroleptic drugs. The proposed role of apoD in CNS lipid transport, combined with the recent evidence that schizophrenia and 30 other neuropsychiatric illnesses are accompanied by abnormalities in lipid metabolism, suggest that apoD could play an important role in the action of clozapine. 101 WO 01/30972 PCT/USOO/29690 EXAMPLE 3 Characterization of CLZ 40 5 Male C57B1/6J mice (20-28 g) were housed as previously described in Example 1. The same experimental paradigm used in Example 1 for clozapine treatment was used for the various analyses described below. Briefly, in the clozapine studies, the control group mice received a single injection of sterile saline (0.1 ml volume), or no injection, 10 and were sacrificed after 45 minutes. The mice subjected to acute clozapine treatment were given a single intraperitoneal injection of clozapine (7.5 mg/kg) and sacrificed after 45 minutes or 7 hours, as described in Example 1. The mice subjected to chronic clozapine treatment received daily subcutaneous injections of clozapine (7.5 mg/kg) for 5 days, 12 days or 14 days. All animals were sacrificed in their cages with CO 2 at the 15 indicated times. Brains were rapidly removed and placed on ice. The striatum, including the nucleus accumbens, were dissected out and placed in ice-cold phosphate-buffered saline. The mRNA was prepared according to the method described in Example 2. For the morphine studies, male C57B1/6J mice (20-28 g) were housed as 20 previously described in Example 1 and divided into the following groups: 1) a control group, in which the mice were subcutaneusly implanted with one placebo pellet upon halothane anaesthesia; 2) an acute morphine group, in which the mice received a morphine intraperitoneal injection of 10 mg/kg; 25 3) a chronic or tolerant group, in which mice were rendered drug-tolerant and dependent by means of subcutaneous implantation of a single pellet containing 75 mg of morphine free base for 3 days; and 4) a withdrawal group, in which the mice rendered tolerant to morphine were injected intraperitoneally with naltrexone 1 mg/kg. Animals were sacrificed in their cages 30 with CO 2 at 72 hours after placebo or morphine pellet implantation, or 4 hours after single injection of morphine, or 4 hours after administration of naltrexone to morphine-tolerant mice. Their brains were rapidly removed. The striatum, including the nucleus accumbens, 102 WO 01/30972 PCT/USOO/29690 and block of tissues containing the amygdala complex were dissected under microscope and collected in ice-cold RNA extraction buffer. The TOGA data shown in Figures 13 and 14 were generated with a 5'-PCR 5 primer (C-G-A-C-G-G-T-A-T-C-G-G-T-T-G-T; SEQ ID NO: 26) paired with the "universal" 3' primer (SEQ ID NO: 23) labeled with 6-carboxyfluorescein (6FAM, ABI) at the 5' terminus. PCR reaction products were resolved by gel electrophoresis on 4.5% acrylamide gels and fluorescence data acquired on ABI377 automated sequencers. Data were analyzed using GeneScan software (Perkin-Elmer). 10 The results of TOGA analysis using a 5' PCR primer with parsing bases C-G-A C-G-G-T-A-T-C-G-G-T-T-G-T (SEQ ID NO: 26) are shown in Figs. 13 and 14, which show PCR products produced from mRNA isolated from the striatum/nucleus accumbens of mice treated with clozapine (Fig. 13) or morphine (Fig. 14). In Fig. 13, the vertical 15 index line indicates a PCR product of about 266 b.p. that is present in control cells, and whose expression decreases in the striatum/nucleus accumbens of mice treated with clozapine for 45 minutes, 7 hours, 5 days, 12 days, and 14 days. The down-regulation of CLZ_40 occurs as early as 45 minutes following clozapine treatment and remains downregulated for at least 14 days. 20 In Fig. 14, the vertical index line indicates a PCR product of about 266 b.p. that is present in control cells, and whose expression differentially regulated in control striatum (PS), acutely treated striatum (AS), withdrawal striatum (WS), control amygdala (PA), acutely treated amygdala (AA), chronically treated amygdala (TA), and withdrawal 25 amygdala (WA). The expression of CLZ_40 product is greater in striatum than in amygdala. Further, CLZ_40 displays chronic-specific or withdrawal-specific regulation in both of these brain regions. In striatum, CLZ_40 is downregulated in withdrawal striatum but not acutely treated striatum. In amygdala, CLZ_40 is slightly upregulated in acutely treated amygdala and increasingly upregulated in chronically treated amygdala 30 and withdrawal amygdala. 103 WO 01/30972 PCT/USOO/29690 Shown in Fig. 15, Northern Blot analysis was performed using mRNA extracted from the striatum/nucleus accumbens of control mice and clozapine-treated mice. Briefly, an agarose gel containing 2pig of poly A enriched mRNA as well as size standards was electrophoresed on a 1.5% agarose gel containing formaldehyde, 5 transferred to a biotrans membrane, and prehybridized for 30 minutes in Expresshyb (Clonetech). An 265 bp insert of CLZ_40 (25-100 ng) was labeled with [a- 2 P]-d CTP by oligonucleotide labeling to specific activities of approximately 5x10 8 cpm/tg and added to the prehybridization solution and incubated 1 hour. Filters were washed to high stringency (0.2 X SSC) (1 X SSC: 0.015 M NaCl and 0.0015 M Na citrate) at 68*C then 10 exposed to Kodak X-AR film (Eastman Kodak, Rochester, NY) for up to 1 week. As shown in Fig. 15, a 9-12 Kb transcript was detected in control and clozapine-treated mice which decreases dramatically after 45 minutes with clozapine treatment and remains down-regulated for at least 14 days. 15 Figure 16 is a graphical representation comparing the results of the clozapine treatment TOGA analysis of clone CLZ_40 shown in Fig. 13 and the clozapine treatment Northern Blot analysis of clone CLZ_40 shown in Figure 15. The Northern Blot was imaged using a phosphoimager to determine the amount of CLZ_40 mRNA in each clozapine-treated sample relative to the amount of mRNA in the control sample. As can 20 be seen, the clozapine treatment TOGA analysis shows correlation with the clozapine treatment Northern Blot analysis. Figure 17A-B is an in situ hybridization analysis, demonstrating CLZ_40 mRNA expression in the mouse brain. In situ hybridization was performed on free-floating 25 sections (25 pM thick). Coronal sections were hybridized at 55*C for 16 hour with an 35 S-labeled, single-stranded antisense cRNA probe of CLZ_40 at 107 cpm/ml. The probe was synthesized from the 3'-ended cDNA TOGA clone using the Maxiscript Transcription Kit (Ambion, Austin, TX). Excess probe was removed by washing with 2 X SSC (I X SSC = 0.015 M NaC1I/0.0015 M Na citrate) containing 14 mM P 30 mercaptoethanol (30 minutes), followed by incubation with 4 pg/ml ribonuclease in 0.5 M NaC1/0.05 M EDTA/0.05 M Tris-HCI, pH 7.5, for 1 hour at 37*C. High stringency 104 WO 01/30972 PCT/USOO/29690 washes were carried out at 55*C for 2 hours in 0.5 X SSC/50% formamide/0.01 M p mercaptoethanol, and then at 68*C for 1 hour in 0.1 X SSC/0.01 M p mercaptoethanol/0.5% sarkosyl. Slices were mounted onto gelatin-coated slides and dehydrated with ethanol and chloroform before autoradiography. Slides were exposed 5 for 1-4 days to Kodak X-AR film and then dipped in Ilford K-5 emulsion. After 4 weeks, slides were developed with Kodak D19 developer, fixed, and counterstained with Richardson's blue stain. Interestingly, CLZ_40 mRNA is specifically expressed in the nucleus accumbens and pyriform cortex (Fig. 17A), and dentate gyrus (Fig. 17B), but is not detected in any other brain regions. 10 At present, CLZ 40 (SEQ ID NO: 12) is of unknown identity. However, the CLZ_40 DST has been PCR amplified and the extended sequence clone of CLZ_40 (SEQ ID NO: 13) matches an EST in the GenBank database (A1509550) as shown in Table 4. The observation that CLZ_40 is down-regulated with clozapine treatment 15 suggests a potential association with the therapeutic effects of clozapine. Furthermore, its highly unique gene expression pattern is like no other gene identified to date, and its presence in the nucleus accumbens may implicate CLZ_40 in a number of functional roles associated with this structure, namely limbic/mental behavior and addiction. 20 Addiction to opiates and other drugs of abuse is a chronic disease of the brain, most likely resulting from molecular and cellular adaptations of specific neurons to repeated exposure to opiates (Leshner, A., Science, 278, 45-47 (1997)). An important neural substrate implicated in the opioid reinforcement and addiction is the mesolimbic system, notably the nucleus accumbens (Everitt, et al, Ann. N.Y. Acad. Sci., 877, 412-438 25 (1999)). All highly addictive drugs, such as opiates, cocaine and amphetamines, produce adaptations in the neural circuitry of the nucleus accumbens, but the precise relationships are unknown. The molecular neuroadaptation which takes place in this structure may offer important insight into the mechanisms of drug addiction. CLZ_40 is a likely candidate for involvement in such mechanisms due to its specific expression in the 30 nucleus accumbens. Elucidation of the biology underlying psychoses and addiction is 105 WO 01/30972 PCT/USOO/29690 key to understanding the underlying causes of such disorders and may lead to the development of more effective treatments, including anti-addiction medications. Furthermore, the behavioral mechanisms associated with addiction reflect 5 mechanisms of learning and memory (White, N., Addiction, 91, 921-949 (1996)). The hippocampal system has long been associated with learning and memory, including forms of conditional associative learning (Sziklas, et al., Hippocampus, 8, 131-137 (1998)), which is the form of learning associated with addiction (Di Chiara, et al., Ann. N. Y Acad. Sci., 877, 461-85 (1999)). The expression of CLZ_40 in the hippocampus suggests that 10 this gene may provide a link with such learning processes. 15 106 WO 01/30972 PCT/USOO/29690 TABLE 1 Seq ID Clone ID Digital Address Control 45 7 Hour 5 Day 12 Day 14 Day (Mspl) minutes AAAA 276 314 189 183 299 292 227 AAAG 91 18 27 34 52 60 35 AAAG 446 135 127 219 245 529 210 AAAG 449 135 173 219 245 775 210 AACA 109 31 17 85 54 51 72 AACA 117 38 30 45 39 72 118 AACA 137 355 205 163 129 111 186 AACA 307 56 58 65 55 134 64 AACG 375 633 450 420 528 968 1015 AACG 498 717 221 349 438 1647 1392 AACT 85 481 139 145 108 281 580 AACT 112 297 162 391 538 330 555 AACT 392 176 267 427 303 296 315 AAGA 309 22 19 42 91 61 36 AAGA 324 37 15 12 83 31 46 AAGC 446 284 212 155 249 318 338 AAGC 498 456 369 309 495 735 862 AAGG 270 169 191 176 243 283 265 AAGG 457 191 129 152 228 269 320 AAGG 497 265 164 208 432 390 512 AAGT 282 75 73 82 84 204 105 AATA 90 47 46 39 74 115 65 60 CLZ_47 AATA 136 817 555 589 297 245 397 AATA 194 70 81 70 133 181 112 AATC 352 108 108 128 144 631 140 AATC 499 49 32 43 67 75 67 AATT 425 38 30 38 37 64 45 ACAA 80 92 67 109 319 353 110 ACAA 122 58 95 107 46 818 98 ACAA 239 117 45 133 49 217 137 ACAC 145 313 365 296 277 750 631 ACAC 273 163 169 262 274 800 338 ACAG 81 167 81 57 137 314 253 ACAG 270 117 94 117 93 236 213 ACAG 296 32 34 71 47 89 62 ACAG 413 39 43 52 43 88 81 ACAG 437 25 20 41 22 55 41 ACAT 94 91 151 149 91 340 195 ACCA 109 318 505 352 289 189 200 107 WO 01/30972 PCT/USOO/29690 ACCA 418 33 28 46 40 65 39 ACCA 422 28 23 44 39 51 39 ACCC 394 32 55 40 38 162 37 ACCC 493 54 42 57 48 93 69 ACCG 90 181 155 184 217 382 208 ACCG 220 169 113 262 189 335 247 ACCG 489 33 30 28 44 63 41 ACCT 119 117 121 47 86 300 164 ACCT 490 78 76 57 120 165 133 ACGA 77 567 133 109 72 1143 1079 ACGA 92 61 56 56 76 195 63 ACGA 292 349 247 165 190 306 148 ACGC 78 243 31 51 236 2323 1676 ACGC 118 1026 737 849 292 442 513 ACGC 210 243 284 293 343 682 735 ACGC 284 27 50 60 195 159 94 ACGC 474 50 91 87 107 190 131 ACGG 264 140 108 117 115 294 172 ACGG 335 245 104 102 110 131 159 59 CLZ_44 ACGG 352 171 407 428 538 683 553 ACGG 382 37 53 113 154 141 103 ACGG 406 114 233 267 217 219 211 ACTA 88 28 37 33 29 219 41 ACTA 199 38 84 48 120 365 66 ACTC 88 64 30 71 124 108 81 ACTC 105 54 121 172 155 352 294 ACTG 266 23 35 116 35 87 44 ACTG 468 148 80 53 74 58 68 ACTT 436 490 549 450 494 435 504 AGAA 104 86 210 143 63 39 106 AGAA 196 62 75 43 85 172 97 AGAA 462 42 29 25 27 64 42 AGAC 410 362 307 538 530 918 442 AGAT 79 41 73 50 64 193 70 AGAT 251 622 622 746 691 562 696 AGAT 295 294 252 263 281 303 263 AGAT 456 603 525 571 639 588 559 AGCA 177 21 38 46 64 163 100 AGCC 295 661 444 517 421 360 475 AGCC 468 112 99 110 165 145 146 AGCG 202 385 349 433 339 334 334 AGCT 95 162 963 1168 2493 3990 1420 AGCT 260 89 78 58 296 86 294 108 WO 01/30972 PCT/USOO/29690 AGGA 426 365 532 720 670 896 802 AGGC 104 46 86 169 163 642 339 AGGG 177 739 251 249 210 174 408 AGGG 242 165 110 192 222 376 293 AGGG 492 35 48 33 46 98 75 AGGG 498 50 47 69 79 155 111 AGGT 99 55 36 55 80 83 61 AGGT 103 29 27 31 50 84 38 AGGT 119 835 719 808 518 466 643 CLZ 3 AGTA 106 657 1677 1883 894 832 1282 AGTC 97 297 229 215 158 111 180 AGTC 178 519 351 238 263 353 269 AGTC 410 65 93 107 85 175 156 AGTG 498 532 851 1476 1209 2196 1092 AGTT 378 48 33 61 40 68 56 ATAA 183 428 319 426 353 915 583 ATAA 225 17 40 39 49 128 82 ATAG 94 52 98 63 343 469 76 ATAG 108 1111 995 933 833 713 869 ATAG 402 495 416 472 546 535 482 ATAT 140 37 20 44 53 45 57 ATCA 90 423 666 451 172 379 180 ATCA 199 774 588 493 335 336 352 ATCT 99 59 43 56 35 125 67 ATCT 392 139 176 287 262 569 226 ATGA 162 91 95 127 239 191 262 ATGC 78 138 91 111 190 466 148 ATGC 124 317 884 743 403 164 317 ATGC 236 15 23 76 7 54 119 ATGC 344 153 108 131 187 217 185 ATGG 96 118 231 173 115 113 305 ATGG 365 15 26 22 25 63 29 ATGT 378 28 47 90 54 108 80 ATGT 383 26 61 78 40 136 63 ATTA 256 36 29 27 46 61 81 ATTA 259 48 54 55 65 75 106 ATTG 88 100 147 147 262 318 114 ATTG 485 22 27 27 26 100 29 ATTT 186 87 60 58 64 190 122 ATTT 189 99 79 74 85 209 127 ATTT 313 79 49 94 86 511 197 ATTT 499 62 80 78 61 265 114 CAAA 423 398 255 395 302 506 434 109 WO 01/30972 PCT/USOO/29690 CAAC 471 87 67 99 85 134 104 CAAC 474 93 77 109 85 151 128 CAAT 319 23 18 22 16 66 30 CACA 253 771 716 598 626 684 579 CACA 348 847 303 241 181 316 342 CACA 374 205 116 308 211 262 175 CACC 98 241 553 402 143 68 363 2 CLZ_5 CACC 201 382 653 727 782 775 903 14 CLZ_6 CACT 169 1576 1400 727 987 933 909 CAGA 119 388 129 217 102 115 119 65 CLZ_52 CAGA 146 737 728 643 511 354 332 CAGA 157 927 820 422 943 533 893 CAGA 214 118 94 79 129 229 163 CAGC 247 508 1511 557 483 531 527 CAGG 129 647 536 588 592 571 493 CATA 172 534 482 447 494 863 625 3 CLZ_8 CATC 98 94 333 253 141 76 212 CATC 135 350 483 606 403 299 464 CATG 78 78 58 56 98 126 217 CATG 197 406 401 421 474 427 318 64 CLZ 51 CATG 247 1740 1436 2195 3089 2713 4020 CATT 420 194 114 155 122 259 214 CATT 429 119 89 96 105 198 141 CATT 432 127 101 106 104 229 157 CCAC 404 28 12 23 37 51 93 CCAG 87 58 29 28 115 100 229 4 CLZ_10 CCAG 104 211 309 353 154 153 262 CCAT 119 122 38 91 35 113 179 CCAT 133 57 45 66 59 95 100 CCAT 296 16 34 7 8 80 56 CCAT 440 56 76 86 104 83 97 CCCC 123 474 860 910 628 277 698 CCCG 243 163 654 354 120 146 129 CCCG 277 218 282 257 310 660 337 CCCG 283 298 261 421 250 779 323 CCCG 454 84 69 115 90 140 102 CCCT 119 107 76 104 146 176 132 CCGC 88 32 231 134 82 843 226 CCGC 93 197 52 18 743 462 367 CCGC 118 2960 2515 1919 1789 1038 540 CCGC 309 153 126 94 78 164 156 CCGG 89 201 406 535 612 446 377 61 CLZ_48 CCGG 94 176 705 527 578 482 702 110 WO 01/30972 PCT/USOO/29690 CCGG 249 563 188 384 393 295 487 CCGG 263 535 275 183 219 309 161 CCGT 169 363 246 408 247 559 398 5 CLZ_12 CCGT 172 765 511 343 347 407 174 63 CLZ 50 CCGT 293 88 57 65 52 426 251 CCGT 350 82 24 91 37 52 100 CCTA 110 174 342 363 204 214 195 CCTA 379 80 89 170 105 192 217 CCTC 382 72 83 88 66 105 110 CCTG 99 283 93 245 1081 319 379 CCTG 130 1413 1995 1550 934 1004 1180 CCTT 104 304 533 768 344 288 0 CGAA 101 66 225 382 71 130 305 CGAC 76 71 45 704 87 174 1047 CGAC 148 1008 1239 1016 884 1043 999 CGAC 480 556 498 421 605 1183 913 CGAC 490 317 250 225 282 531 473 CGAG 273 212 98 136 89 96 136 CGAG 450 122 122 101 173 230 181 CGAT 78 322 85 178 293 484 420 CGAT 95 42 40 62 80 94 50 CGAT 98 48 62 67 68 124 52 CGAT 105 97 59 45 199 206 151 CGAT 268 770 202 374 593 519 478 CGAT 496 170 164 127 196 147 146 CGCA 88 592 249 355 696 542 854 CGCA 334 1071 1923 1725 1333 1445 1438 CGCA 472 218 306 294 365 312 406 CGCG 82 61 115 148 377 254 133 CGCG 85 32 115 60 275 248 133 CGCG 111 49 236 266 826 778 323 CGCG 371 27 37 72 44 101 56 CGCT 118 905 634 948 855 668 542 CGCT 341 22 29 39 11 62 23 CGGC 87 66 89 149 216 198 150 CGGC 110 311 620 1099 292 124 687 CGGG 85 259 928 777 314 252 437 CGGG 102 35 35 175 93 365 99 CGGG 109 34 28 63 65 112 96 CGGG 135 100 203 120 91 434 537 CGGG 402 116 116 170 205 226 178 CGGG 490 59 69 116 116 142 100 CGGT 142 207 147 171 201 301 322 111 WO 01/30972 PCT/USOO/29690 6 CLZ_15 CGGT 217 174 116 130 91 87 83 CGGT 476 46 30 29 41 60 53 CGTC 342 71 87 121 79 393 92 CGTG 124 346 240 174 115 144 168 CGTG 234 346 131 129 105 71 119 CGTG 306 796 1334 1296 1163 1164 1114 CGTT 81 42 91 35 129 186 74 CGTT 245 169 161 216 168 402 185 CTAA 268 125 133 121 157 151 201 58 CLZ_43 CTAA 461 120 131 146 185 397 220 CTAC 93 90 73 124 101 146 106 CTAC 359 184 161 249 238 357 258 CTAG 91 48 29 64 113 142 175 CTAG 97 360 331 395 116 102 537 15 CLZ_16 CTAG 171 412 247 167 119 181 142 CTAT 190 61 41 67 59 89 74 49 CLZ_17 CTCA 206 567 522 466 306 370 239 CTCA 313 39 19 47 36 51 55 CTCG 140 90 94 293 259 663 605 CTCG 218 1262 450 734 340 124 208 CTCG 331 59 28 84 49 88 104 CTCG 490 352 257 320 376 616 504 CTCG 498 258 152 234 315 597 488 CTCT 137 503 422 462 762 965 828 CTCT 142 1146 797 1258 1620 1881 1685 CTGA 115 29 30 42 30 130 55 62 CLZ_49 CTGA 450 127 173 228 279 258 265 CTGC 116 0 449 479 212 188 0 57 CLZ 18 CTGC 320 0 60 83 99 104 0 CTGG 84 102 54 62 90 117 126 CTGG 183 269 195 328 321 308 1166 CTTA 86 49 24 69 48 73 52 CTTA 132 58 45 58 60 97 58 CTTA 378 297 350 416 443 747 450 CTTA 494 31 24 39 24 56 44 CTTA 499 10 29 45 42 69 52 CTTC 77 26 30 49 58 64 45 CTTG 83 792 397 700 601 967 1173 CTTG 176 119 75 200 187 192 229 GAAC 78 35 17 117 36 36 51 GAAG 93 122 348 230 116 116 183 GAAG 148 552 569 635 454 343 560 GAAG 196 363 237 448 447 223 350 112 WO 01/30972 PCT/USOO/29690 GAAG 223 44 31 51 63 71 101 GAAG 226 44 31 51 62 71 81 GAAG 231 18 15 30 31 71 85 GACG 79 26 20 38 47 57 62 GACG 97 597 409 195 127 214 160 GACG 423 187 294 260 280 377 377 GACT 155 117 111 137 201 241 147 GAGG 103 136 175 399 79 90 139 GAGG 248 227 82 85 120 112 117 GAGT 367 302 382 345 369 355 326 GATA 345 15 33 31 50 94 30 GATC 95 81 170 177 112 67 130 GATC 356 34 35 67 48 108 42 GATG 300 375 310 202 280 270 293 GATT 91 50 18 32 41 40 55 GCAA 90 211 210 261 303 206 194 GCAA 269 222 90 150 140 218 237 GCAC 92 63 82 119 59 416 266 GCAC 186 282 238 186 308 203 156 GCAT 121 229 260 229 149 166 222 GOAT 439 19 25 28 34 57 35 GCCA 112 189 312 216 134 102 213 GOCA 240 49 47 22 27 119 68 GCCC 79 60 42 40 62 89 101 GCCC 121 62 42 39 57 96 212 GCCC 294 695 144 403 428 422 469 67 CLZ_56 GCCC 324 202 648 578 521 512 802 GCCG 139 57 36 128 115 146 87 GCCG 144 78 39 71 52 101 139 GCCT 84 122 68 102 166 150 165 GCCT 118 403 671 853 366 337 489 GCCT 126 561 294 305 328 188 246 GCGA 180 235 1349 636 733 1018 1159 GCGA 293 1031 312 375 643 332 335 68 CLZ_57 GCGC 325 35 61 60 75 104 95 GCGG 77 65 79 91 73 193 78 GCGG 127 51 50 52 107 161 130 GCGG 254 413 167 190 231 214 251 GCGG 269 842 133 372 326 480 586 GCGG 471 93 130 112 129 149 147 GCGT 140 117 55 78 115 189 159 GCGT 168 701 465 504 599 429 405 GCGT 309 498 282 77 186 71 139 113 WO 01/30972 PCT/USOO/29690 GCTA 109 388 639 619 320 267 550 GCTA 132 990 829 1198 735 669 968 GCTA 223 898 532 586 525 812 522 16 CLZ_22 GCTA 292 444 169 168 171 182 154 GCTC 174 100 26 34 57 109 114 GCTC 202 785 866 512 626 949 593 GCTC 326 752 666 793 862 890 1479 GCTG 78 103 116 427 446 587 312 GCTG 120 1694 2136 2033 1141 1119 1652 GCTG 172 31 31 116 154 114 74 GCTT 233 43 20 62 23 51 63 GGAA 434 49 114 93 142 230 125 GGAC 231 683 585 478 510 254 236 GGAC 472 62 50 62 68 112 120 GGAG 221 423 239 203 217 250 248 GGAG 372 836 772 775 1052 913 641 GGAT 223 1048 1430 1425 1632 944 1461 GGCA 305 155 124 206 194 280 164 7 CLZ_24 GGCA 393 303 544 393 608 725 842 GGCC 113 334 371 479 204 175 240 GGCC 134 838 720 633 537 668 608 GGCC 324 114 115 211 157 238 301 GGCC 418 40 12 32 28 26 52 GGCG 113 235 158 129 129 130 101 GGCG 136 97 61 76 59 125 145 GGCG 315 292 238 445 464 495 366 50 CLZ_26 GGCT 129 491 544 423 199 169 321 69 CLZ_60 GGCT 169 467 563 335 704 1233 1055 GGCT 176 127 173 164 410 407 230 GGGA 172 91 97 67 144 112 183 GGGA 377 307 157 252 269 263 255 GGGC 214 59 62 85 66 252 255 GGGC 286 27 23 34 29 60 71 GGGG 81 670 1443 1269 1095 2164 1645 GGGT 91 63 68 104 267 143 91 GGTA 128 265 198 142 124 153 146 GGTA 184 1209 969 875 1109 836 941 51 CLZ_28 GGTA 257 1016 872 549 492 539 422 GGTG 139 992 884 936 801 733 811 GGTT 100 12 17 35 32 41 94 GTAA 257 86 36 105 119 75 98 GTAC 107 815 975 1034 821 751 1057 GTAG 244 260 237 294 349 736 282 114 WO 01/30972 PCT/USOO/29690 GTAG 459 113 137 168 239 351 199 GTAG 459 113 137 168 239 351 199 GTAG 471 75 68 76 103 172 99 GTCC 87 448 256 218 325 193 176 GTCC 124 111 443 155 139 104 160 GTCC 187 1253 1031 1066 1018 891 778 GTCC 413 28 29 42 35 61 42 GTCG 176 55 58 79 190 130 126 GTCG 228 3085 2559 3211 3000 3470 3051 GTCT 84 19 28 30 43 128 35 GTGC 87 58 106 159 316 867 410 GTGG 125 1407 1734 1004 1276 1047 1475 GTGG 147 821 314 343 174 188 188 GTGG 458 45 22 41 35 26 33 GTTC 491 90 236 206 175 240 176 GTTG 93 156 129 90 93 150 88 GTTG 114 20 37 44 58 75 78 GTTG 378 66 35 74 59 80 73 GTTT 260 49 24 33 42 56 49 GTTT 336 37 42 40 36 139 126 GTTT 339 31 37 40 34 156 108 GTTT 495 36 23 34 54 58 50 TAAA 84 27 25 46 37 60 37 TAAC 114 38 32 50 48 65 41 TAAC 222 411 367 454 384 216 229 TAAC 450 678 538 407 452 753 669 TAAG 386 210 334 126 421 702 301 TACA 119 42 49 73 98 111 103 TACA 129 282 242 227 197 206 180 TACA 200 801 493 438 442 477 324 TACC 99 132 141 88 51 18 102 TACC 129 185 160 327 486 457 247 TACC 169 122 72 83 103 179 255 TACC 344 88 71 89 79 104 183 17 CLZ_32 TACG 274 181 206 160 187 255 578 TACT 151 94 34 53 44 97 132 8 CLZ 33 TACT 188 184 278 1200 581 339 347 TACT 386 36 50 70 56 104 88 TAGA 125 41 88 152 95 195 106 TAGA 134 286 263 214 194 146 152 TAGA 242 32 9 26 37 142 51 TAGC 186 1357 1306 1263 1125 959 889 TAGC 411 56 68 76 76 142 123 115 WO 01/30972 PCT/USOO/29690 TAGC 415 50 60 40 66 127 87 TAGC 464 183 184 166 133 129 106 TAGG 250 461 166 238 189 306 257 TAGT 81 213 160 178 286 473 369 TAGT 97 271 144 246 309 537 299 TATA 98 115 183 488 127 99 230 TATA 382 37 36 49 44 113 39 72 CLZ_65 TATC 159 434 327 334 404 701 2760 TATC 262 119 154 204 168 826 154 71 CLZ_62 TATG 290 135 103 59 121 37 52 TATG 446 201 229 389 325 462 328 9 CLZ 34 TATT 89 156 623 509 129 186 314 TATT 112 50 38 182 101 122 50 TATT 119 43 16 25 52 40 43 TATT 230 403 42 24 31 35 103 TATT 272 59 43 59 57 131 88 TATT 354 44 36 63 42 147 99 TCAA 447 44 38 39 26 85 49 TCAC 134 836 1637 842 57 1228 1047 TCAC 212 777 567 742 688 573 552 TCAC 289 1707 1138 1116 842 943 1123 TCAG 84 56 68 205 125 148 108 MAT 88 88 145 178 409 401 430 18 CLZ_36 TCAT 349 2478 380 1155 1425 903 1832 70 CLZ_64 TCAT 391 314 216 421 391 554 699 TCAT 473 45 22 38 39 53 47 TCCA 106 150 71 193 91 179 385 TCCA 222 400 303 362 613 787 616 TCCA 435 68 78 56 57 241 71 TCCA 439 54 78 56 61 174 71 10 CLZ_37 TCCC 97 381 1687 1532 720 673 1083 TCCC 148 1050 865 963 700 639 685 TCCG 120 0 832 774 566 649 653 TCCG 185 0 311 292 223 206 259 TCCT 98 577 621 882 925 1258 1741 TCCT 144 492 551 427 580 313 410 TCCT 166 740 488 588 605 421 473 TCCT 275 72 20 77 52 108 133 TCGA 255 533 263 431 473 614 575 TCGA 370 167 148 178 194 215 229 TCGC 196 229 155 214 97 412 311 TCGC 328 465 545 856 482 674 773 TCGT 326 32 32 95 33 85 34 116 WO 01/30972 PCT/USOO/29690 TCTA 80 49 65 184 75 563 231 TCTA 217 39 50 160 325 212 84 TCTC 143 341 256 203 262 229 141 TCTT 155 93 80 96 91 252 110 TGAA 240 542 390 530 667 552 540 TGAC 193 1029 566 798 752 902 1048 19 CLZ_42 TGAC 328 194 216 199 314 475 303 TGAT 97 45 44 23 72 158 85 TGAT 138 608 468 542 442 467 498 11 CLZ_38 TGCA 109 339 554 561 473 736 395 TGCA 185 137 83 67 160 382 346 TGCC 163 271 347 93 330 958 407 TGCC 185 1164 1680 573 1081 1145 992 TGCC 343 604 628 832 675 889 1068 TGCG 77 188 156 495 125 366 403 TGCG 111 36 50 76 225 167 155 TGGA 93 173 157 202 253 545 240 TGGA 108 1941 294 2077 1692 1853 2640 TGGA 154 823 1504 1481 1370 1122 673 TGGA 277 50 23 54 56 103 93 TGGA 308 31 32 52 51 149 84 TGGC 105 634 538 630 818 1092 669 TGGC 113 377 259 371 510 524 415 TGGC 160 156 213 282 223 460 320 TGGC 266 468 451 365 280 207 270 TGGC 276 73 81 59 81 251 274 TGGC 494 98 43 27 58 88 122 TGGG 93 33 65 48 55 228 583 TGGG 271 241 591 580 426 642 607 TGGT 103 76 25 97 93 132 236 TGGT 114 339 537 421 221 204 231 TGGT 122 119 145 180 135 341 182 TGGT 158 465 286 403 324 267 348 TGGT 330 666 673 726 770 701 753 TGTA 121 1021 1596 1727 1052 696 1206 TGTA 169 1562 681 624 801 880 753 TGTC 84 160 250 216 410 510 399 TGTC 109 711 704 686 276 149 466 TGTG 315 71 56 83 35 125 73 TGTG 393 430 313 425 528 419 664 TGTG 450 573 554 698 819 1166 654 TGTT 114 335 752 657 875 794 838 TGTT 119 703 .1167 .993 1666 1824 1251 117 WO 01/30972 PCT/USOO/29690 TGTT 453 138 226 333 307 324 287 TTAA 88 149 109 181 377 239 326 TTAA 194 369 115 230 262 391 313 TTAA 312 335 177 159 199 136 167 TTAC 174 287 294 137 192 196 180 TTAG 104 52 51 54 44 112 65 TTAT 106 41 22 50 232 53 44 TTAT 338 486 777 852 875 816 884 TTCC 96 97 140 133 130 370 135 TTCC 104 51 31 109 67 94 78 TTGA 117 20 28 34 38 63 60 TTGC 119 57 52 67 73 117 75 TTGC 299 151 114 68 60 65 59 TTGG 209 704 1160 894 921 857 1215 TTGG 466 60 47 46 71 103 68 12 CLZ_40 TTGT 266 200 52 75 82 67 115 TTGT 302 38 33 72 48 69 79 TTGT 483 53 87 120 110 140 60 TTTA 249 174 32 103 46 55 85 TTTC 85 31 44 34 89 369 100 TTTC 107 50 37 20 65 91 68 TTTC 118 633 721 715 303 257 483 TTTC 153 188 168 113 141 142 270 TTTC 171 663 642 709 704 801 589 TTTC 226 26 31 22 63 63 86 TTTC 277 566 324 375 327 381 278 118 WO 01/30972 PCT/USOO/29690 Zc) 0 CdC -c~ ItI0t C>. CN tn0 0 0 C) CC eq 00 ON 0 0s 0 ~O00 -~ CC er 00. 00) 0A 00 .z o C0~ ~~n -ON cn 0 . K Uc-n co' ~0 -- ~0 ~ UC - ll - cc = 0C "It 0. 0. 9 : " C C 0000 -A Nl N7 . C 0M N - O 0 -n 0- U~1 iiI 119 WO 01/30972 PCT/USOO/29690 0l0 w Cl C l I I NN C1 kN 00 C -4 tn 0 D C) Cl Nl 000 00 _t e" C's 00 en Cl->~C aN kn L) e 00 = v0 0 ,0 0 w a~ 00 0 0J w 0. - - Dc I.. 30 u~ _. en cn U0 q~C ~ 0 0 Ul- kn~ A 'n >e u0 -0. Pm CP - 4 w0 0 00 w/2 C/ C *) 00 ON ON- C 00 11N 0 N O - _00 ON -1 -- cl - en 00 0 C Qn en enm O- C r 00 ON - - - -- 120 WO 01/30972 PCT/USOO/29690 ONON 0 ~ tn C* ON 0 o ~(OO N 11 00N 0 0 In NOr N a., I o I I 00 Nl a., 00 . 0 Lflt 00 - -. O 00 C6 aN I N O 'O S 00 00 N It 00 N N 00 r- N C0 ON ON ON ON, ON ON ON ON*N, u 000 r O - t 0 Cc~ 0N Nq *i -C 0 q ~ H 0 i 00 tC.~O 0 'ITS~ 0O 0i 0 04~' cq So 0 H-H Hl En 00 Hz 0l 0 0 Hn H> U -~0 C)\ ( 0 0 O F- u u \C N1 00 ON N000 0 121 WO 01/30972 PCT/USOO/29690 r- t 00 o> W 0 M -E 00 C > 0 00 Q~". 0 -0 ' n A - s Cl aA 0 n 1. - 0~ 0'.0 - n Zo a- (n a- I 40 rjn U 6 0- 00 eq 0,~ 0 0 U 0 0 (D dH c a* N * C '1 Q -w C14 - cn MI c~-~ 00~.O ~ u uZ c o 0~' 0 0 1 - C 1 -u~ u- Cl Cl u u w U n t- 0 0 H 0 - Cl0 122 WO 01/30972 PCT/USOO/29690 00 "0 0 .H 1 .I 00 et ) 0 co 0 Co in C) \- r 00 0~ 0 a CI0 H ~tn cn ou 123 -.

WO 01/30972 PCT/USOO/29690 H 00 CNCLenI N N H ci H 0 Q Li L < H H H H < < H < < 00 0 < H H S0 H H H 0 < 0 eH cO L Li 0 Q < HO .9 0 <~ 0 H Li L 0 0 S<H < 0 0 0 0- Li LiO i Li < 0 0 O L Li< L 0 H H H SH Li H 0 0 Li L Li LiLiLi Li 0 H 0 0 0 0 0 00 0 0 00 0 0 00 0 H H H H H H H H 0 00 0 0 00 0 H H H H H H H H 0 0 0 0 0 00 0 uU 0 u 00 (D 0 Q! QD 0Q e -2 o I 04o u o u u u 0 0 (D~ I rA e o ~En o 0o 0 0H bD C >0 E Zi i "Ii , 0 .0s -h =1 0 0. 4 c ~0 =1 00o-6 -: " ; W~ 'n 0 0 E Li Li 00 En 6 tr , > 4)0 f 00 - cri CA 112 WO 01/30972 PCT/USOO/29690 00 0 0 U E < <U U 0 0 0 0 C) H H 0 0 0 0 o) H 0 C) 0 0 H ) .- 0 H 0 0 C)o H 0 C) 0 SH C) < H 0 U U U H C< C)0 0 < H U U U U *H H H H H C) 0 0 0-0 0 0 0 H1H < H H H H H H H H H H U _ c~ ~ I C I 00 U U cl C.' c tu N N NN N 25 Ho H0 w ) CZ) C) C) Cc t12 WO 01/30972 PCT/USOO/29690 ~0 O F-0 0 H u~ 0 H H H < H <0 0 C) 0< <UQ 0 m H 0 < 0 o C) 0 * C) < C) H H < 0 .i 0 C) < 0 H H H & H 0 00o <0o H u~ Q o 0 H C) C < 0 < 0 3 C) < < 0 < 0H ,e < ) O H H C)Q & H H C) C < H H H H H H H H u u 0 0 00 (D 0D SHH HH HHH H . < << 00 .ono -0 og c en u u o-z u ) -- EA o ccO c z CIO o u om A4 Fn p E ~Cn t 2 0 UrO z . o. - ~ ~ C, - s ,CA - .

0 UI.0 q . Wo tn 00o e o o m 0 .2 O\ F .o cn < o 0 C4o u < < (D& H U Io eq C <0 0 H 0 0 N N N N N N N N 126 WO 01/30972 PCT/USOO/29690 00 0 - m N N 00 00 00 00 00 00 u < -< < < < 0 U F-u - 0 0 i 0 u u< < D F- < u 4 u Q << 0< 0 0 0 0 SH < 0 < < 0 O < F 0 Q U F U u U U 0 G U O 0 0 F u oa u e u 00 0 0(0 0 0C0 co -'4 o?? < <<H< H < < < < < U U U U U U U < << < < < al 0 0 - r2 000 0 0 0 0 - ~ -AA 00 0 0 0 o en w ~ C) Uw s.O t; I Ont as6 06r ON ON hCr cj 004 r- -4 (u w 00 F- O nU 0 U F l F- U o U U 0 U O0 0' F - N 0O N 0 e' 0) 3. w 0<t, 0r -. C .~ N N N NN N> U U U U U U U 1 2 .tw u C u8 o u uo s uo u ( O.Nx 'e 273 3 ,3 0 .2 bg WO 01/30972 PCT/USOO/29690 00 00 u H u 0 u (-C) S0 U 00 w 0- 0 0 0 Z U UZ 0 oN m e - "oLn = ' u 4 u u 128 WO 01/30972 PCT/USOO/29690 0 00 t U 110 zH 0 0 CA 0 cn m 0 d w 0 00 u wUo ' H1 Hu u~ ~) -129 WO 01/30972 PCT/USOO/29690 u u u0 U0 u0 u0 u 00 Z Z 0 U UU 00 0 00 00 ~ CC) 2 0 v) 11 1) 00) :3 0 r.000 -bo L)0 0 0 r-0 00 - Cu ~130 WO 01/30972 PCT/USOO/29690 z culu 1 00 *00 .n 00'6 o uo 00 Cun 0 0 kff) C> -,z -0 r U u u c .13 WO 01/30972 PCT/USOO/29690 EXAMPLE 4 Characterization of CLZ 34 5 Male C57Bl/6J mice (20-28 g) were housed as previously described in Example 1. The same experimental paradigm used in Example 1 for clozapine treatment was used for the TOGA analyses described below. 10 The TOGA data shown in Figure 18 was generated with a 5'-PCR primer (C G-A-C-G-G-T-A-T-C-G-G-T-A-T-T; SEQ ID NO: 27) paired with the "universal" 3' primer (SEQ ID NO: 23) labeled with 6-carboxyfluorescein (6FAM, ABI) at the 5' terminus. PCR reaction products were resolved by gel electrophoresis on 4.5% acrylamide gels and fluorescence data acquired on AB1377 automated sequencers. 15 Data were analyzed using GeneScan software (Perkin-Elmer). The results of TOGA analysis using a 5' PCR primer with parsing bases C-G A-C-G-G-T-A-T-C-G-G-T-A-T-T (SEQ ID NO: 27) are shown in Figure 18, which shows PCR products produced from mRNA isolated from the striatum/nucleus 20 accumbens of mice treated with clozapine for various lengths of time as described in Example 1. In Fig. 18, the vertical index line indicates a PCR product of about 89 b.p. that is present in control cells, and whose expression in the striatum/nucleus accumbens of mice treated with clozapine is differntially regulated with acute treatment versus chronic treatment. CLZ_34 is upregulated with clozapine treatment 25 at 45 minutes and 7 hours, but decreases to control level by day 5 and remains at about control level for as long as 12 days, showing a slight increase at day 14. In situ analysis performed using CLZ_34 as a probe revealed that CLZ_34 is expressed ubiquitously throughout the brain (data not shown). 30 CLZ_34 corresponds with GenBank sequence U08262, which is identified as a rat N-methyl-D-aspartate receptor/NMDAR1-2a subunit (NMDAR1). The NMDAR1 receptor is a glutamate receptor involved in the processes underlying learning and memory. In addition, numerous studies show that blockade of glutamate actions by noncompetitive (e.g. MK801 and dextromethorphan) and competitive (e.g. 132 WO 01/30972 PCT/USOO/29690 LY274614) NMDA receptor antagonists blocks or reduces the development of morphine tolerance following long term opiate administration (Trujillo et al., Science, 251, 85-87, (1991); Elliott et al., Pain, 56, 69-75 (1994); Wiesenfeld-Hallin, Z., Neuropsychopharm., 13, 347-56 (1995)). The early change in the level of expression 5 of CLZ_34 which has high homology with an NMDA receptor is interesting in view of the ability of NMDA antagonists to block the development of tolerance to opioids. EXAMPLE 5 10 Figure 19 shows the consensus sequence from the computer generated assembly of the following 4 sequences A1415388: Soares mouse p3NMF19.5 Mus musculus cDNA clone IMAGE:350746 3', mRNA sequence; A1841003: UI-M-AMO ado-e-04-0-UI.sl NIHBMAPMAM Mus musculus cDNA clone UI-M-AMO-ado-e 04-0-U 3', mRNA sequence; A1413353: Soares mouse embryo NbME13.5 14.5 Mus 15 musculus cDNA IMAGE:356159 3', mRNA sequence; A1425991: Soares mouse embryo NbME13.5 14.5 Mus musculus cDNA IMAGE:426077 3', mRNA sequence. (SEQ ID NO: 53) Figure 20 shows the sequence of the EST AF006196: Mus musculus 20 metalloprotease-disintegrin MDC15 mRNA, complete cds. (SEQ ID NO: 54) Figure 21 shows the consensus sequence from the computer generated assembly of the following 3 sequences: C86593: Mus musculus fertilized egg cDNA 3'-end sequence, clone J0229E09 3', mRNA sequence; A1428410: Life Tech mouse 25 embryo 13 5dpc 10666014 Mus musculus cDNA clone IMAGE:553802 3', mRNA sequence; A1561814: Stratagene mouse skin (#937313) Mus musculus cDNA clone IMAGE:1227449 3', mRNA sequence. (SEQ ID NO: 55). 30 EXAMPLE 6 Characterization of CLZ 44 Male C57Bl/6J mice (20-28 g) were housed as previously described in Example 1. The same experimental paradigm used in Example 1 for clozapine treatment was used for the TOGA analyses. The TOGA data was generated with a 5' 35 PCR primer (C-G-A-C-G-G-T-A-T-C-G-G-A-C-G-G; SEQ ID NO:96) paired with 133 WO 01/30972 PCT/USOO/29690 the "universal" 3' primer (SEQ ID NO: 23) labeled with 6-carboxyfluorescein (6FAM, ABI) at the 5' terminus. PCR reaction products were resolved by gel electrophoresis on 4.5% acrylamide gels and fluorescence data acquired on AB1377 automated sequencers. Data were analyzed using GeneScan software (Perkin-Elmer). 5 As shown in Table 1, the results of TOGA analysis indicate that CLZ_44 is slightly up-regulated by clozapine treatment. Tables 2 and 3 show that CLZ_44 is an EST isolated from mouse kidney. In further characterization of CLZ_44, northern blot analyses were performed to determine the pattern of expression in the 10 striatum/nucleus accumbens after 2 weeks of treatment of control mice, clozapine treated mice, haloperidol-treated mice, and ketanserin-treated mice (Figure 22). Ketanserin is a 5HT2A/2c - selective antagonist, and is used to determine serotonorgic involvement in these drug effects. 15 Briefly, an agarose gel containing 2ptg of poly A enriched mRNA as well as size standards was electrophoresed on a 1.5% agarose gel containing formaldehyde, transferred to a biotrans membrane, and prehybridized for 30 minutes in Expresshyb (Clonetech). A CLZ_44 insert (25-100 ng) was labeled with [a- 2 P]-d CTP by oligonucleotide labeling to specific activities of approximately 5x10 8 cpm/tg and 20 added to the prehybridization solution and incubated 1 hour. Filters were washed to high stringency (0.2 X SSC) (1 X SSC: 0.015 M NaCl and 0.0015 M Na citrate) at 68*C then exposed to Kodak X-AR film (Eastman Kodak, Rochester, NY) for up to 1 week. Figure 22 is a graphical representation of the described northern blot analyses. As shown, after 2 weeks of treatment, CLZ_44 was up-regulated with haloperidol and 25 ketanserin, but not clozapine. This suggests that both dopamines D2 and 5HT2M2C receptors are involved in CLZ_44 expression regulation. The lack of effect of clozapine may indicate that antagonism at other receptors (i.e. 5HT 3 , D4, D1) may override the effects of D2, 5HT 2 receptors. 30 EXAMPLE 7 Characterization of CLZ 38 Male C57Bl/6J mice (20-28 g) were housed as previously described in Example 1. The same experimental paradigm used in Example 1 for clozapine 134 WO 01/30972 PCT/USOO/29690 treatment was used for the TOGA analyses. The TOGA data was generated with a 5' PCR primer (C-G-A-C-G-G-T-A-T-C-G-G-T-G-C-A; SEQ ID NO: 97) paired with the "universal" 3' primer (SEQ ID NO: 23) labeled with 6-carboxyfluorescein (6FAM, ABI) at the 5' terminus. PCR reaction products were resolved by gel 5 electrophoresis on 4.5% acrylamide gels and fluorescence data acquired on AB1377 automated sequencers. Data were analyzed using GeneScan software (Perkin-Elmer). Tables 2 and 3 show that CLZ_38 is an oligodendrocyte-specific protein mRNA. In further characterization of CLZ_38, northern blot analyses were 10 performed to determine the pattern of expression in the striatum/nucleus accumbens of control miceand mice treated with clozapine for 45 minutes, 7 hours, 5 days, and 2 weeks (Figure 23). Briefly, an agarose gel containing 2pg of poly A enriched mRNA as well as 15 size standards was electrophoresed on a 1.5% agarose gel containing formaldehyde, transferred to a biotrans membrane, and prehybridized for 30 minutes in Expresshyb (Clonetech). A CLZ_38 insert (25-100 ng) was labeled with [a- 2 P]-d CTP by oligonucleotide labeling to specific activities of approximately 5x 108 cpm/pg and added to the prehybridization solution and incubated 1 hour. Filters were washed to 20 high stringency (0.2 X SSC) (1 X SSC: 0.015 M NaCl and 0.0015 M Na citrate) at 68*C then exposed to Kodak X-AR film (Eastman Kodak, Rochester, NY) for up to 1 week. Figure 23 is a graphical representation of the described northern blot analyses. As shown, the pattern of CLZ_38 expression in clozapine-treated animals was similar to the pattern observed with TOGA analysis. 25 EXAMPLE 8 Characterization of CLZ 16 Male C57Bl/6J mice (20-28 g) were housed as previously described in Example 1. The same experimental paradigm used in Example 1 for clozapine 30 treatment was used for the TOGA analyses. The TOGA data was generated with a 5' PCR primer (C-G-A-C-G-G-T-A-T-C-G-G-C-T-A-G; SEQ ID NO: 97) paired with the "universal" 3' primer (SEQ ID NO: 23) labeled with 6-carboxyfluorescein (6FAM, ABI) at the 5' terminus. PCR reaction products were resolved by gel 135 WO 01/30972 PCT/USOO/29690 electrophoresis on 4.5% acrylamide gels and fluorescence data acquired on AB1377 automated sequencers. Data were analyzed using GeneScan software (Perkin-Elmer). As shown in Table 1, the results of TOGA analysis indicate that CLZ_16 is 5 slightly down-regulated by clozapine treatment. Tables 2 and 3 show that CLZ_16 is an arm-repeat protein. In further characterization of CLZ_16, in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_16 were performed to show the pattern of CLZ_16 mRNA expression in mouse anterior brain (24B) and posterior brain (24A). Control mice and mice treated with 7.5 mg/kg 10 clozapine were sacrificed after two weeks. In situ hybridization was performed on free-floating sections (25 piM thick). Coronal sections were hybridized at 55*C for 16 hour with an 35 S-labeled, single-stranded antisense cRNA probe of CLZ_16 at 10 7 cpm/ml. 15 The probe was synthesized from the 3'-ended cDNA TOGA clone using the Maxiscript Transcription Kit (Ambion, Austin, TX). Excess probe was removed by washing with 2 X SSC (I X SSC = 0.015 M NaCI/0.0015 M Na citrate) containing 14 mM p-mercaptoethanol (30 minutes), followed by incubation with 4 pig/ml ribonuclease in 0.5 M NaCI/0.05 M EDTA/0.05 M Tris-HCl, pH 7.5, for 1 hour at 370 20 C. High stringency washes were carried out at 55*C for 2 hours in 0.5 X SSC/50% formamide/0.01 M p-mercaptoethanol, and then at 68*C for 1 hour in 0.1 X SSC/0.01 M P-mercaptoethanol/0.5% sarkosyl. Slices were mounted onto gelatin-coated slides and dehydrated with ethanol and chloroform before autoradiography. Slides were exposed for 1-4 days to Kodak X-AR film and then dipped in Ilford K-5 emulsion. 25 After 4 weeks, slides were developed with Kodak D19 developer, fixed, and counterstained with Richardson's blue stain. As shown in Figure 24A and B, CLZ_16 mRNA is expressed ubiquitously throughout mouse brain. Figure 24A shows dense labelling in the cortex and 30 surrounding the hippocampal formation as well as moderate labelling in the dorsal thalamus and posterior brain. Figure 24B shows uniform labelling throughout. 136 WO 01/30972 PCT/USOO/29690 EXAMPLE 9 Characterization of CLZ 17 Male C57Bl/6J mice (20-28 g) were housed as previously described in Example 1. The same experimental paradigm used in Example 1 for clozapine 5 treatment was used for the TOGA analyses. The TOGA data was generated with a 5' PCR primer (C-G-A-C-G-G-T-A-T-C-G-G-C-T-C-A; SEQ ID NO: 99) paired with the "universal" 3' primer (SEQ ID NO: 23) labeled with 6-carboxyfluorescein (6FAM, ABI) at the 5' terminus. PCR reaction products were resolved by gel electrophoresis on 4.5% acrylamide gels and fluorescence data acquired on AB1377 10 automated sequencers. Data were analyzed using GeneScan software (Perkin-Elmer). As shown in Table 1, the results of TOGA analysis indicate that CLZ_17 is slightly down-regulated by clozapine treatment. Table 4 shows that CLZ_17 matches several ESTs isolated from mouse tissue. In further characterization of CLZ_ 17, in 15 situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_17 were performed to show the pattern of CLZ_17 mRNA expression in mouse sections from anterior (25B) and posterior regions of the brain (25A). In situ hybridization was performed on free-floating sections (25 piM thick) 20 taken from control mice and mice treated with 7.5 mg/kg clozapine for 2 weeks. Coronal sections were hybridized at 55*C for 16 hour with an 35 S-labeled, single stranded antisense cRNA probe of CLZ_17 at 107 cpm/ml. The probe was synthesized from the 3'-ended cDNA TOGA clone using the Maxiscript Transcription Kit (Ambion, Austin, TX). Excess probe was removed by washing as previously 25 described in Example 8. Slices were mounted onto gelatin-coated slides and dehydrated with ethanol and chloroform before autoradiography. Slides were exposed for 1-4 days to Kodak X-AR film and then dipped in Ilford K-5 emulsion. After 4 weeks, slides were developed with Kodak D19 developer, fixed, and counterstained with Richardson's blue stain. 30 Figure 25A-B shows an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_17, showing the pattern of CLZ_17 mRNA expression in a coronal sections from posterior (25A) and anterior (25B) 137 WO 01/30972 PCT/USOO/29690 regions of mouse brain. As shown, CLZ_17 mRNA is expressed in the cortex, hippocampus, striatum, and amygdala. 5 EXAMPLE 10 Characterization of CLZ 24 Male C57B1/6J mice (20-28 g) were housed as previously described in Example 1. The same experimental paradigm used in Example 1 for clozapine 10 treatment was used for the TOGA analyses. The TOGA data was generated with a 5' PCR primer (C-G-A-C-G-G-T-A-T-C-G-G-G-G-C-A; SEQ ID NO: 100) paired with the "universal" 3' primer (SEQ ID NO: 23) labeled with 6-carboxyfluorescein (6FAM, ABI) at the 5' terminus. PCR reaction products were resolved by gel electrophoresis on 4.5% acrylamide gels and fluorescence data acquired on AB1377 15 automated sequencers. Data were analyzed using GeneScan software (Perkin-Elmer). As shown in Table 1, the results of TOGA analysis indicate that CLZ_24 is up-regulated by clozapine treatment. Tables 2 and 3 show that CLZ_24 is an EST isolated from rat tissue. In further characterization of CLZ_24, in situ hybridization 20 analysis using an antisense cRNA probe directed against the 3' end of CLZ_24 were performed to show the pattern of CLZ_24 mRNA expression in mouse anterior brain (26B) and posterior brain (26A) In situ hybridization was performed on free-floating sections (25 IM thick) 25 obtained from comtrol mica nd mice treated with 7.5 mg/kg clozapine for 2 weeks. Coronal sections were hybridized at 55 0 C for 16 hour with an 35 S-labeled, single stranded antisense cRNA probe of CLZ_24 at 107 cpm/ml. The probe was synthesized from the 3'-ended cDNA TOGA clone using the Maxiscript Transcription Kit (Ambion, Austin, TX). Excess probe was removed by washing as previously 30 described in Example 8. Slices were mounted onto gelatin-coated slides and dehydrated with ethanol and chloroform before autoradiography. Slides were exposed for 1-4 days to Kodak X-AR film and then dipped in Ilford K-5 emulsion. After 4 weeks, slides were developed with Kodak D19 developer, fixed, and counterstained with Richardson's blue stain. 138 WO 01/30972 PCT/USOO/29690 Figure 26A-B shows an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_24, showing the pattern of CLZ_24 mRNA expression in a coronal section through the hemispheres (26A) and cross 5 section through the brainstem (26B) in mouse brain. As shown, CLZ_24 mRNA is ubiquitously expressed in the cortex. EXAMPLE 11 10 Characterization of CLZ 26 Male C57B1/6J mice (20-28 g) were housed as previously described in Example 1. The same experimental paradigm used in Example 1 for clozapine treatment was used for the TOGA analyses. The TOGA data was generated with a 5' PCR primer (C-G-A-C-G-G-T-A-T-C-G-G-G-G-C-T; SEQ ID NO: 101) paired with 15 the "universal" 3' primer (SEQ ID NO: 23) labeled with 6-carboxyfluorescein (6FAM, ABI) at the 5' terminus. PCR reaction products were resolved by gel electrophoresis on 4.5% acrylamide gels and fluorescence data acquired on AB1377 automated sequencers. Data were analyzed using GeneScan software (Perkin-Elmer). 20 As shown in Table 1, the results of TOGA analysis indicate that CLZ_26 is slightly down-regulated by clozapine treatment. Table 4 shows that CLZ_26 is a metalloprotease-disintegrin MDC15 mRNA. In further characterization of CLZ_26, in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_26 were performed to show the pattern of CLZ_26 mRNA expression in 25 mouse anterior brain (27B) and posterior brain (27A). In situ hybridization was performed on free-floating coronal sections (25 pLM thick) with an 35 S-labeled, single-stranded antisense cRNA probe of CLZ_26 using the methods described in the above examples. 30 Figure 27A-B is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_26, showing the pattern of CLZ_26 mRNA expression in a coronal section of the hemispheres at the level of hippocampal 139 WO 01/30972 PCT/USOO/29690 formation (27A) and coronal section of the hemispheres at the level of striatum (27B) in mouse brain. As shown, CLZ_26 mRNA is ubiquitously expressed in the cortex. 5 EXAMPLE 12 Characterization of CLZ 28 Male C57Bl/6J mice (20-28 g) were housed as previously described in Example 1. The same experimental paradigm used in Example 1 for clozapine treatment was used for the TOGA analyses. The TOGA data was generated with a 5' 10 PCR primer (C-G-A-C-G-G-T-A-T-C-G-G-G-G-T-A; SEQ ID NO: 102) paired with the "universal" 3' primer (SEQ ID NO: 23) labeled with 6-carboxyfluorescein (6FAM, ABI) at the 5' terminus. PCR reaction products were resolved by gel electrophoresis on 4.5% acrylamide gels and fluorescence data acquired on AB1377 automated sequencers. Data were analyzed using GeneScan software (Perkin-Elmer). 15 As shown in Table 1, the results of TOGA analysis indicate that CLZ_28 is down-regulated by clozapine treatment. Table 4 shows that CLZ_28 matches several ESTs isolated from mouse tissue. In further characterization of CLZ_28, in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of 20 CLZ_28 were performed to show the pattern of CLZ_28 mRNA expression in mouse anterior brain (28B) and posterior brain (28A). In situ hybridization was performed on free-floating coronal sections (25 p.M thick) with an 35 S-labeled, single-stranded antisense cRNA probe of CLZ_28 using 25 the methods described in the above examples. Figure 28A-B is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_28, showing the pattern of CLZ_28 mRNA 30 expression in a coronal section through the hemispheres at the level of hippocampus (28A) and coronal section through the posterior region of hemispheres (28B) in mouse brain. As shown in Figure 28A and B, CLZ_28 mRNA is expressed ubiquitously in the cortex. 140 WO 01/30972 PCT/USOO/29690 EXAMPLE 13 Characterization of CLZ 3 Male C57B1/6J mice (20-28 g) were housed as previously described in Example 1. The same experimental paradigm used in Example 1 for clozapine 5 treatment was used for the TOGA analyses. The TOGA data was generated with a 5' PCR primer (C-G-A-C-G-G-T-A-T-C-G-G-A-G-T-A; SEQ ID NO: 94) paired with the "universal" 3' primer (SEQ ID NO: 23) labeled with 6-carboxyfluorescein (6FAM, ABI) at the 5' terminus. PCR reaction products were resolved by gel electrophoresis on 4.5% acrylamide gels and fluorescence data acquired on AB1377 10 automated sequencers. Data were analyzed using GeneScan software (Perkin-Elmer). As shown in Table 1, the results of TOGA analysis indicate that CLZ_3 is up regulated by clozapine treatment. Tables 2 and 3 show that CLZ_3 is a seine protease HTRA mRNA. In further characterization of CLZ_3, in situ hybridization 15 analysis using an antisense cRNA probe directed against the 3' end of CLZ_3 were performed to show the pattern of CLZ_3 mRNA expression in mouse anterior brain (29B) and posterior brain (29A). In situ hybridization was performed on free-floating coronal sections (25 tM 20 thick) with an 35 S-labeled, single-stranded antisense cRNA probe of CLZ_3 using the methods described in the above examples. Figure 29A-B is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_3, showing the pattern of CLZ_3 mRNA 25 expression in a coronal section through the hemispheres at level of hippocampus (29A) and cross section through midbrain (29B) in mouse brain. As shown in Figure 29A and B, CLZ_3 mRNA is expressed in the cortex, thalamus, hippocampus, striatum, and amygdala. 30 EXAMPLE 14 Characterization of CLZ 34 Male C57B1/6J mice (20-28 g) were housed as previously described in Example 1. The same experimental paradigm used in Example 1 for clozapine 141 WO 01/30972 PCT/USOO/29690 treatment was used for the TOGA analyses. The TOGA data was generated with a 5' PCR primer (C-G-A-C-G-G-T-A-T-C-G-G-T-A-T-T; SEQ ID NO: 103) paired with the "universal" 3' primer (SEQ ID NO: 23) labeled with 6-carboxyfluorescein (6FAM, ABI) at the 5' terminus. PCR reaction products were resolved by gel 5 electrophoresis on 4.5% acrylamide gels and fluorescence data acquired on AB1377 automated sequencers. Data were analyzed using GeneScan software (Perkin-Elmer). As shown in Table 1, the results of TOGA analysis indicate that CLZ_34 is up-regulated by clozapine treatment. Tables 2 and 3 show that CLZ_34 is an N 10 methyl-D-aspartate receptor NMDAR1-2a subunit mRNA. In further characterization of CLZ_34, in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_34 were performed to show the pattern of CLZ_34 mRNA expression in mouse anterior brain (30B) and posterior brain (30A). 15 In situ hybridization was performed on free-floating coronal sections (25 pM thick) with an 35 S-labeled, single-stranded antisense cRNA probe of CLZ_34 using the methods described in the above examples. 20 Figure 30A-B is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_34, showing the pattern of CLZ_34 mRNA expression in a coronal section through the hemispheres at the level of hippocampus (30A) and cross section through the midbrain (30B) in mouse brain. As shown in Figure 30A and B, CLZ_34 mRNA is ubiquitously expressed. 25 EXAMPLE 15 Characterization of CLZ 43 Male C57Bl/6J mice (20-28 g) were housed as previously described in Example 1. The same experimental paradigm used in Example 1 for clozapine 30 treatment was used for the TOGA analyses. The TOGA data was generated with a 5' PCR primer (C-G-A-C-G-G-T-A-T-C-G-G-C-T-A-A; SEQ ID NO: 104) paired with the "universal" 3' primer (SEQ ID NO: 23) labeled with 6-carboxyfluorescein (6FAM, ABI) at the 5' terminus. PCR reaction products were resolved by gel 142 WO 01/30972 PCT/USOO/29690 electrophoresis on 4.5% acrylamide gels and fluorescence data acquired on AB1377 automated sequencers. Data were analyzed using GeneScan software (Perkin-Elmer). As shown in Table 1, the results of TOGA analysis indicate that CLZ_43 is 5 up-regulated by clozapine treatment. Tables 2 and 3 show that CLZ_43 matches an EST isolated from mouse tissue that matches oxysterol binding protein family member. In further characterization of CLZ_43, in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_43 were performed to show the pattern of CLZ_43 mRNA expression in mouse anterior brain (31 C), 10 midbrain (31A), and posterior brain (31B). In situ hybridization was performed on free-floating coronal sections (25 pLM thick) with an 35 S-labeled, single-stranded antisense cRNA probe of CLZ_43 using the methods described in the above examples. 15 Figure 3 1A-C is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_43, showing the pattern of CLZ_43 mRNA expression in coronal sections of the hemispheres showing in the cortex, and intense lebelling in the striatum (31A-C) in mouse brain. Comparison with brain sections 20 obtained from control mice showed that CLZ_43 expression is increased approximately 10-fold by chronic treatment (2 weeks) with clozapine. Following the cloning of the mouse DST CLZ_43, a BLAST analysis was performed. A human homology was identified as a 5556 b.p. GenBank entry 25 (AB040884, also known as KIAA1451). An oligonucleotide was chosen from this sequence and used to isolate the remaining 5' end of the human gene from an adult human brain cDNA plasmid library. Using the method described below, a 1717 b.p. cDNA clone (SEQ ID NO:103) was isolated that overlaps the human sequence. This clone provides an additional (novel) 512 b.p. at the 5' end of the GenBank entry. 30 Sequence analysis suggests the position of the methionine start codon for the open reading frame is at base 562 of the 1717 b.p. clone (SEQ ID NO: 108). The open reading frame of the 1717 b.p. clone encodes a 385 amino acid peptide (SEQ ID NO: 108, SEQ ID NO: 109). 143 WO 01/30972 PCT/USOO/29690 The following methods were used to isolate the 1717 b.p. cDNA clone. The target pool was a cDNA plasmid library created from adult human brain RNA. The oligonucleotide sequence used for hybridization was 5' - AAC AAG TCC GTC CTG GCA TGG-3' (SEQ ID NO:88). The clone was isolated using the methods prescribed 5 by the manufacturer of the GeneTrapper kit (Life Technologies, Inc.). Capture oligonucleotide were prepared by end-labeling the oligonucleotide with biotin-14 dCTP using terminal deoxynucloetidyl transferase. The cDNA plasmid pool was converted from double-stranded cDNA to single-stranded cDNA through the specific action of GeneIl protein and exonuclease III. The single-stranded cDNA pool was 10 combined with the end-labelled oligonucleotide and hybridization was allowed to occur at room temperature for 30 minutes. The reaction was then mixed with strepavidin-coated magnetic beads. The single-stranded cDNA plasmids that hybridized to the oligonucleotide were purified using a magnet to retain the magnetic beads in the reaction tube while all of the unbound components were washed away. 15 The single-stranded plasmid DNA was released from the oligonucleotide and repaired back into a double-stranded plasmid using a fresh sample of the capture oligonucleotide and DNA polymerase. The repaired plasmids were transformed into bacteria and plated on an agar plate. The following day, bacterial colonies were individually picked and grown overnight. Plasmid DNA was prepared from these 20 mini-preparations and subjected to sequence analysis. Homology matches with a human genome database have identified 7 exons spread across more than 22,000 b.p. Further it has been determined that CLZ_43 maps to chromosome 12, which is not a chromosome previously linked to 25 schizophrenia. The sequence data reveals that the open reading frame encodes a protein of 472 amino acids (SEQ ID NO: 110). Comparison with protein databases indicate that the protein is novel and is a member of a class of proteins that binds lipids, especially oxysterols. 30 The observation that, of thousands of proteins expressed by the striatum, apoD and a novel oxysterol binding protein are among the few modulated by neuroleptic drugs strengthens the hypothesis that schizophrenia is a disease of brain sterol homeostasis, and thus may have etiologies as diverse as atherosclerosis. The brain has by far more cholesterol and 24S-hydroxysterol than any organ other than the 144 WO 01/30972 PCT/USOO/29690 adrenal glands, and the special importance of the membrane activities of neurons and their myelinating cells are self-evident. The lipid bilayer of the membrane is made up of glycerolphopholipids and cholesterol, and variations in composition and hydrocarbon chain saturation state determine membrane order and fluidity. These 5 properties affect the binding of extrinsic membrane proteins and, thus, second messenger signaling. As we have shown previously, a large percentage of the mRNAs highly enriched in the striatum encode proteins that regulate second messenger signaling along the inner membrane. Thus, a panneural or panorganismic disruption in lipid metabolism might manifest first as a striatal disease. As of now, 10 this is a somewhat impressionistic concept. Working out the nature of the neuroleptic drug effects on membrane properties may bring the issue into greater focus. EXAMPLE 16 Characterization of CLZ 44 15 Male C57B1/6J mice (20-28 g) were housed as previously described in Example 1. The same experimental paradigm used in Example 1 for clozapine treatment was used for the TOGA analyses. The TOGA data was generated with a 5' PCR primer (C-G-A-C-G-G-T-A-T-C-G-G-A-C-G-G; SEQ ID NO: 105) paired with the "universal" 3' primer (SEQ ID NO: 23) labeled with 6-carboxyfluorescein 20 (6FAM, ABI) at the 5' terminus. PCR reaction products were resolved by gel electrophoresis on 4.5% acrylamide gels and fluorescence data acquired on AB1377 automated sequencers. Data were analyzed using GeneScan software (Perkin-Elmer). As shown in Table 1, the results of TOGA analysis indicate that CLZ_44 is 25 up-regulated by clozapine treatment. Tables 2 and 3 show that CLZ_44 matches an EST isolated from mouse tissue. In further characterization of CLZ_44, in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_44 were performed to show the pattern of CLZ_44 mRNA expression in mouse anterior brain (32A) and posterior brain (32B). 30 In situ hybridization was performed on free-floating coronal sections (25 pLM thick) with an 3 5 S-labeled, single-stranded antisense cRNA probe of CLZ_44 using the methods described in the above examples. 145 WO 01/30972 PCT/USOO/29690 Figure 32A-B is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_44, showing the pattern of CLZ_44 mRNA expression in a coronal section showing labelling in the hippocampus, hypothalamus, and temporal cortex (32A) and coronal section showing cortical labelling (32B) in 5 mouse brain. EXAMPLE 17 Characterization of CLZ 64 Male C57B1/6J mice (20-28 g) were housed as previously described in 10 Example 1. The same experimental paradigm used in Example 1 for clozapine treatment was used for the TOGA analyses. The TOGA data was generated with a 5' PCR primer (C-G-A-C-G-G-T-A-T-C-G-G-T-C-A-T; SEQ ID NO: 106) paired with the "universal" 3' primer (SEQ ID NO: 23) labeled with 6-carboxyfluorescein (6FAM, ABI) at the 5' terminus. PCR reaction products were resolved by gel 15 electrophoresis on 4.5% acrylamide gels and fluorescence data acquired on AB1377 automated sequencers. Data were analyzed using GeneScan software (Perkin-Elmer). As shown in Table 1, the results of TOGA analysis indicate that CLZ_64 is up-regulated by chronic clozapine treatment. Tables 2 and 3 show that CLZ_64 20 matches an EST isolated from mouse tissue and shares homolgy with mitochondrial enoyl-CoA hydratase mRNA. In further characterization of CLZ_64, in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_64 were performed to show the pattern of CLZ_64 mRNA expression in mouse anterior brain (33B) and mid-brain (33A). 25 In situ hybridization was performed on free-floating coronal sections (25 piM thick) with an 35 S-labeled, single-stranded antisense cRNA probe of CLZ_64 using the methods described in the above examples. 30 Figure 33A-B is an in situ hybridization analysis using an antisense cRNA probe directed against the 3' end of CLZ_64, showing the pattern of CLZ_64 mRNA expression in different coronal sections of the hemispheres in mouse brain. As shown in Figure 33A and B, CLZ_64 mRNA is ubiquitously expressed. 146

Claims (54)

1. An isolated nucleic acid molecule comprising a polynucleotide chosen 5 from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO: 49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO: 57, 10 SEQ ID NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72 and SEQ ID NO:107.

2. An isolated polypeptide encoded by a polynucleotide chosen from the 15 group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO: 49, SEQ ID NO:50, SEQ ID NO:51, SEQ ID NO:52, SEQ ID NO: 57, SEQ ID 20 NO:58, SEQ ID NO:59, SEQ ID NO:60, SEQ ID NO:61, SEQ ID NO:62, SEQ ID NO:63, SEQ ID NO:64, SEQ ID NO:65, SEQ ID NO:66, SEQ ID NO:67, SEQ ID NO:68, SEQ ID NO:69, SEQ ID NO:70, SEQ ID NO:71, SEQ ID NO:72 and SEQ ID NO:107.

3. An isolated polypeptide of SEQ ID NO:109. 25

4. An isolated polypeptide of SEQ ID NO: 110.

5. An isolated nucleic acid molecule comprising a polynucleotide at least 95% identical to the isolated nucleic acid molecule of claim 1.

6. An isolated nucleic acid molecule at least ten bases in length that is hybridizable to the isolated nucleic acid molecule of claim 1 under stringent 30 conditions.

7. An isolated nucleic acid molecule encoding the polypeptide of claim 2.

8. An isolated nucleic acid molecule encoding a fragment of the polypeptide of claim 2. 147 WO 01/30972 PCT/USOO/29690

9. An isolated nucleic acid molecule encoding a polypeptide epitope of the polypeptide of claim 2.

10. The polypeptide of claim 2 wherein the polypeptide has biological activity. 5

11. An isolated nucleic acid encoding a species homologue of the polypeptide of claim 2.

12. The isolated nucleic acid molecule of claim 1, wherein the nucleotide sequence comprises sequential nucleotide deletions from either the 5' end or the 3'end. 10

13. A recombinant vector comprising the isolated nucleic acid molecule of claim 1.

14. A recombinant host cell comprising the isolated nucleic acid molecule of claim 1.

15. A method of making the recombinant host cell of claim 14. 15

16. The recombinant host cell of claim 14 comprising vector sequences.

17. The isolated polypeptide of claim 2, wherein the isolated polypeptide comprises sequential amino acid deletions from either the C-terminus or the N terminus.

18. An isolated antibody that binds specifically to the isolated polypeptide 20 of claim 2.

19. An isolated antibody that binds specifically to the isolated polypeptide of claim 3.

20. An isolated antibody that binds specifically to the isolated polypeptide of claim 4. 25

21. The isolated antibody of claims 16, 17 or 18 wherein the antibody is a monoclonal antibody.

22. The isolated antibody of claims 16, 17 or 18 wherein the antibody is a polyclonal antibody.

23. A recombinant host cell that expresses the isolated polypeptides of 30 claim 2, 3 or 4.

24. An isolated polypeptide produced by the steps of: (a) culturing the recombinant host cell of claim 14 under conditions such that said polypeptide is expressed; and (b) isolating the polypeptide. 148 WO 01/30972 PCT/USOO/29690

25. A method for preventing, treating, modulating, or ameliorating a medical condition, comprising administering to a mammalian subject a therapeutically effective amount of the polypeptide of claims 2, 3 or 4, or the polynucleotide of claim 1. 5

26. The method of claim 25 wherein the medical condition is a neuropsychiatric disorder.

27. A method for preventing, treating, modulating, or ameliorating a medical condition comprising administering to a mammalian subject a therapeutically effective amount of the antibody of claims 18, 19 or 20. 10

28. The method of claim 27 wherein the medical condition is a neuropsychiatric disorder.

29. A method of diagnosing a pathological condition or a susceptibility to a pathological condition in a subject comprising: (a) determining the presence or absence of a mutation in the polynucleotide 15 of claim 1; and (b) diagnosing a pathological condition or a susceptibility to a pathological condition based on the presence or absence of said mutation.

30. The method of claim 29 wherein the pathological condition is a neuropsychiatric disorder. 20 30. A method of diagnosing a pathological condition or a susceptibility to a pathological condition in a subject comprising detecting an alteration in expression of a polypeptide encoded by the polynucleotide of claim 1, wherein the presence of an alteration in expression of the polypeptide is indicative of the pathological condition or susceptibility to the pathological condition. 25

31. The method of claim 30 wherein the alteration in expression is an increase in the amount of expression or a decrease in the amount of expression.

32. The method of claim 30 wherein the pathological condition is a neuropsychiatric disorder.

33. The method of claim 32 wherein the method further comprises the 30 steps of: obtaining a first biological sample from a patient suspected of having a neuropsychiatric disorder and obtaining a second sample from a suitable comparable control source; (a) determining the amount of at least one polypeptide encoded by a polynucleotide of claim lin the first and second sample; and 149 WO 01/30972 PCT/USOO/29690 (b) comparing the amount of the polypeptide in the first and second samples; wherein a patient is diagnosed as having a neuropsychiatric disorder if the amount of the polypeptide in the first sample is greater than or less than the amount of 5 the polypeptide in the second sample.

34. The use of the polynucleotide of claim 1 or polypeptide of claims 2, 3 or 4 for the manufacture of a medicament for the treatment of a neuropsychiatric disorder.

35. The use of the antibody of claims 18, 19 or 20 for the manufacture of a 10 medicament for the treatment of a neuropsychiatric disorder.

36. A method for identifying a binding partner to the polypeptide of claims 2, 3 or 4 comprising: (a) contacting the polypeptide of claim 2, 3 or 4 with a binding partner; and (b) determining whether the binding partner effects an activity of the 15 polypeptide.

37. The gene corresponding to the cDNA sequence of the isolated nucleic acid of claim 1.

38. A method of identifying an activity of an expressed polypeptide in a biological assay, wherein the method comprises: 20 (a) expressing the polypeptide of claims 2, 3 or 4 in a cell; (b) isolating the expressed polypeptide; (c) testing the expressed polypeptide for an activity in a biological assay; and (d) identifying the activity of the expressed polypeptide based on the test results. 25

39. A substantially pure isolated DNA molecule suitable for use as a probe for genes regulated by neuroleptics, chosen from the group consisting of the DNA molecules identified in Table 1, having a 5' partial nucleotide sequence and length as described by their digital address, and having a characteristic regulation pattern by neuroleptics. 30

40. A kit for detecting the presence of the polypeptide of the claims 2, 3 or 4 in a mammalian tissue sample comprising a first antibody which immunoreacts with a mammalian protein encoded by a gene corresponding to the polynucleotide of claim 1 or with a polypeptide encoded by the polynucleotide of claim 2, 3 or 4 in an amount sufficient for at least one assay and suitable packaging material. 150 WO 01/30972 PCT/USOO/29690

41. A kit of claim 40 further comprising a second antibody that binds to the first antibody.

42. The kit of claim 41 wherein the second antibody is labeled.

43. The kit of claim 42 wherein the label comprises enzymes, 5 radioisotopes, fluorescent compounds, colloidal metals, chemiluminescent compounds, phosphorescent compounds, or bioluminescent compounds.

44. A kit for detecting the presence of a genes encoding an protein comprising a polynucleotide of claim 1, or fragment thereof having at least 10 contiguous bases, in an amount sufficient for at least one assay, and suitable 10 packaging material.

45. A method for detecting the presence of a nucleic acid encoding a protein in a mammalian tissue sample, comprising the steps of: (a) hybridizing a polynucleotide of claim 1 or fragment thereof having at least 10 contiguous bases, with the nucleic acid of the sample; and 15 (b) detecting the presence of the hybridization product.

46. A method of diagnosing a neuropsychiatric disorder or a susceptibility to a neuropsychiatric disorder in a subject comprising: (a) determining the presence or absence of a mutation in apolipoprotein D polynucleotide; and 20 (b) diagnosing a neuropsychiatric disorder or a susceptibility to a neuropsychiatric disorder based on the presence or absence of said mutation.

47. A method of diagnosing a neuropsychiatric disorder or a susceptibility to a neuropyschiatric disorder in a subject comprising: (a) determining the presence or amount of expression of apolipoprotein D 25 polypeptide in a biological sample; and (b) diagnosing a neuropsychiatric disorder or a susceptibility to a neuropsychiatric disorder based on the presence or amount of expression of the apolipoprotein D polypeptide.

48. The method of claims 46 or 47 wherein the neuropsychiatric disorder 30 is schizophrenia.

49. The method of claims 46 or 47 wherein the neuropsychiatric disorder is bipolar disorder.

50. A method of diagnosing a neuropsychiatric disorder or a susceptibility to a neuropsychiatric disorder in a subject comprising: 151 WO 01/30972 PCT/USOO/29690 (a) determining the presence or absence of a mutation in the polynucleotide or polynucleotide fragment of SEQ ID NO: 2 and (b) diagnosing a neuropsychiatric disorder or a susceptibility to a neuropsychiatric disorder based on the presence or absence of said mutation. 5

51. A method of diagnosing a neuropsychiatric disorder or a susceptibility to a neuropsychiatric disorder in a subject comprising: (a) determining the presence or amount of expression of the polypeptide comprising an amino acid sequence at least 95% identical to a polypeptide fragment of a translation of SEQ ID NO: 2 in a biological sample; and 10 (b) diagnosing a neuropsychiatric disorder or a susceptibility to a neuropsychiatric disorder based on the presence or amount of expression of the polypeptide.

52. The method of claims 50 or 51 wherein the neuropsychiatric disorder is schizophrenia. 15

53. The method of claims 50 or 51 wherein the neuropsychiatric disorder is bipolar disorder.

54. The method of claims 50 or 51wherein the neuropsychiatric disorder is addiction-related behavior. 20 152