WO1997031015A1 - Recepteurs de l'obesite du rat et nucleotides codant ces recepteurs - Google Patents

Recepteurs de l'obesite du rat et nucleotides codant ces recepteurs Download PDF

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Publication number
WO1997031015A1
WO1997031015A1 PCT/US1997/002397 US9702397W WO9731015A1 WO 1997031015 A1 WO1997031015 A1 WO 1997031015A1 US 9702397 W US9702397 W US 9702397W WO 9731015 A1 WO9731015 A1 WO 9731015A1
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Prior art keywords
rat
gene
assay
receptor
nucleic acid
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PCT/US1997/002397
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English (en)
Inventor
John W. Hess
C. Thomas Caskey
Qingyun Liu
Michael S. Phillips
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Merck & Co., Inc.
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Publication date
Priority claimed from GBGB9608473.6A external-priority patent/GB9608473D0/en
Application filed by Merck & Co., Inc. filed Critical Merck & Co., Inc.
Priority to JP53025497A priority Critical patent/JP2002515739A/ja
Priority to EP97905980A priority patent/EP0922052A4/fr
Publication of WO1997031015A1 publication Critical patent/WO1997031015A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
    • C07K14/72—Receptors; Cell surface antigens; Cell surface determinants for hormones

Definitions

  • This invention relates to rat ob receptor proteins, to DNA and RNA sequences encoding them, and to assays using rat receptor proteins.
  • Two different ob alleles have been identified: one mutation causes the premature termination of the leptin peptide resulting in a truncated protein, and the other mutation changes the transcriptional activity of the obesity ⁇ ob) gene, resulting in a reduced amount of circulating leptin.
  • leptin Although the synthesis of leptin occurs in the adipocyte, its ability to decrease food intake and increase metabolic rate appears to be mediated centrally by the hypothalamus. Injection of recombinant leptin into the third ventricle of the brain elicits a similar response as peripheral administration of leptin. Furthermore, the recent cloning of the human receptor for the leptin, the ob-receptor (OB-R), reveals that it is transcribed in the hypothalamus (Tartaglia et al. 1995, Cell 83:1263-1271 ; Stephens et al. 1995, Nature 3.11: 530-532).
  • OB-R ob-receptor
  • The/ ⁇ mutation is a recessive allele that arose spontaneously in the 13M rat strain and was first reported in 1961 (Zucker et al. 1961. 7. Heredity 52: 275-278.
  • the onset of obesity in the fa/fa Zucker rat is at 5-7 weeks of age and progresses with age.
  • the mature fatty rat is approximately twice the weight of lean litter mates and over 40% of its body weight is adipose tissue (Zucker et al. 1962, Proc. Soc. Exp. Biol. Med. 1 10: 165-171 ; Zucker et al. 1963, J.
  • the fa/fa Zucker rat exhibits hypercholesterolemia, hyperlipemia, and hyperglycemia and has been used extensively as an animal model for human cardiovascular disease and diabetes. Most of the fatty Zucker rat colonies have been maintained by outbreeding in order to retain heterozygousity at as many loci as possible. However, certain stocks have been inbred to produce animals such as the Zucker diabetic fatty (ZDF) rat which exhibits a more profound diabetic phenotype than the outbred fa/fa Zucker rat (Clark, et al. 1983, Proc. Soc. Exp. Biol. Med. 173: 68-75).
  • ZDF Zucker diabetic fatty
  • the fa mutation maps to rat chromosome 5 in a region that is syntenic with the db allele on mouse chromosome 4 (Truett, et al. 1991 , Proc. Natl. Acad. Sci. 88: 7806-7809).
  • This observation in conjunction with the similar phenotypes of the fa/fa rat and the dbldb mouse, led to the proposal that the/ ⁇ gene was the rat homologue of the db gene.
  • Higher resolution genetic mapping supports the contention that the fa mutation is located in the gene encoding the rat OB-R (Chua et al. Science 271 : 994).
  • FIGURE 1 is the amino acid sequence of the rat OB- receptor.
  • FIGURE 2 is the cDNA sequence of the rat OB-receptor.
  • FIGURE 3 is a table of primers used for the PCR reactions detailed in the Examples.
  • FIGURE 4 shows the gels demonstrating the analysis of the A ⁇ 80 to C mutation identified in the OB-receptor from hypothalamic cDNA and genomic DNA obtained from lean and fa/fa rats.
  • FIGURE 5 compares the amino acid sequence between human cytokine receptor gpl 30 (Humgp 130), the mouse OB-R (MousOBR), human OB-R (HumOBR) and lean rat OB-R (RatOBR).
  • the numbering refers to the location in the protein, and the cytokine motif GXWSXWS can be seen.
  • Substantially free from associated rat membrane proteins means that the rat receptor protein is not in physical contact with any rat membrane proteins.
  • Substantially purified rat OB-receptor means that the rat receptor protein is at least 90% and preferably at least 95% pure.
  • Wild type means that the gene or protein is substantially the same as that found in a rat which is not considered to have a mutation for that gene or protein. It is also referred to as “lean” throughout the specification and claims. 'fa” means that the gene or protein is substantially the same as that found in a rat homologous for the fatty mutation.
  • nucleic acid or amino acid sequence means either it is the same as the reference sequence, or if not exactly the same, contains changes which do not affect its biological activity or function.
  • the/ ⁇ and wild type rat OB-R genes differ by only one nucleotide, they are not considered “substantially the same” as the biological activity and functions of their encoded proteins are very different.
  • the rat OB-R is a member of the cytokine receptor family. Motifs that are characteristic of the cytokine receptors such as the motif WSXWS (where W is the amino acid residue tryptophan, S is the amino acid residue serine and X is any amino acid.) were found to be conserved in the rat OB-R.
  • One aspect of this invention is the molecular cloning of a rat OB-R.
  • the nucleotide sequence for the rat OB-R from both lean and falfa rat hypothalamic cDNA was determined and compared.
  • the falfa rat there was a single nucleotide change, an A to C at nucleotide 880 resulting in an amino acid change at glutamine 269 to proline.
  • the mutation introduces an Msp I site (CCGG) that was utilized to genotype a number of lean control and fatty animals. The results indicate that the mutation is tightly linked to the/ ⁇ allele.
  • rat OB-R alleles i.e. the OB-R containing a glutamine 269 and the allele containing proline 269 are part of this invention, as are all nucleic acids which can encode them.
  • the nucleotide sequence of the wild type rat OB-R cDNA obtained in accordance with this invention has 3650 nucleotides, as shown in FIGURE 2.
  • This DNA sequence contains an open reading frame from nucleotide 75 to 3653 that encodes a protein of 1 162 amino acids.
  • the open reading frame extending from nucleotide 75 to 3653 makes up one aspect of this invention.
  • the wild type and fa receptor proteins contain an extracellular, a transmembrane domain.
  • the extracellular domain extends from amino acids 1 -830; the transmembrane domain is from amino acids 839-860; and the cytoplasmic domain is from amino acids 860-1 162.
  • This invention also includes proteins which lack one or more of these domains. Such deleted proteins are useful tn assays for identifying ligands and their binding activity.
  • Amino acids 1-28 form a signal sequence; thus the mature proteins extend from amino acids 28- 1 162.
  • the mature proteins form yet another aspect of this invention. This differs from the signal sequence of 1 -22 reported for mouse and human OB-r; this may be explained by the use of a different analysis program.
  • rat OB-R nucleotide sequence is 93% identical to the mouse OB-R and 81 % identical to the human OB-R sequences.
  • the deduced amino acid sequence of the rat OB receptor is 93% identical to the mouse and 76% identical to the human OB-R.
  • the size of the open reading frame of the rat OB-receptor of this invention (1 162 amino acids) is similar to that of the human OB- R (1 165 amino acids) reported by Toriaglla et al. 1995, Cell 83: 1 -20. Both the rat OB-R of this invention and the human OB-R contain a large cytoplasmic domain. In contrast, the mouse OB-receptor of 894 amino acids has a relatively short cytoplasmic domain.
  • One of the most notable and surprising aspects of this invention is that there is only a single nucleotide difference between the wild type rat cDNA and the falfa rat cDNA for the OB-R.
  • PCR fragments obtained from falfa cDNA were sequenced.
  • a single nucleotide change relative to the lean cDNA sequence was observed in the hypothalamus.
  • An A to C transversion at bp 880 results in an amino acid change of glutamine to proline at amino acid residue 268. Every tissue examined in the falfa rat was found to be homozygous for this A to C mutation at nucleotide 880.
  • the A to C change in the sequence introduces a Mspl restriction endonuclease site (CCGG) into the sequence, and this is the basis of an assay for presence of the mutation.
  • CCGG Mspl restriction endonuclease site
  • another aspect of this invention is an assay to determine the genotype of a OB-R DNA, suspected of having an A to C mutation at bp 880, comprising digesting the OB-R DNA with Mspl, and comparing the restriction products so producted.
  • the assay comprises generating PCR products of the OB-R DNA, digesting the PCR products with Mspl, and comparin the restriction products so produced with those obtained from . at containing a wild-type OB-R gene.
  • the gene from a rat which has a wild-type OB-R will yield two restriction products, 1774 and 289 bp long.
  • the gene from the / ⁇ rat will have three restriction products: 747, 1027 and 289 bp long. These are easily observed using standard gel techniques.
  • the OB-R gene can be introduced into virtually any host cell using known vectors. Preferred host cells include E. coli as well as mammalian and yeast cell lines.
  • the OB-R gene may be present in the vector in its native form, or it may be under the control of a heterologous promoter, and if desired, one or more enhancers, or other sequences known to regulate transcription or translation.
  • the host cell containing the OB-R gene is cultured, and the OB-R gene is expressed. After a suitable period of time the OB-R protein may be harvested from the cell using conventional separation techniques.
  • a further aspect of this invention is the use of rat OB-R in assays to identify OB-R ligands.
  • a ligand binds to the OB-R, and in vivo may or may not result in an activation of the receptor.
  • Ligands may be agonists of the receptor (i.e. stimulate its activity), antagonists (inhibit its activity) or they may bind with little or no effect upon the receptor activity.
  • the rat OB-R of this invention is exposed to a putative ligand, and the amount of binding is measured.
  • the amount of binding may be measured in many ways; for example, a ligand or the OB-R being investigated may be labeled with a conventional label (such as a radioactive or fluorescent label) and then put in contact with the OB-R under binding conditions. After a suitable time, the unbound ligand is saparated from the OB-R and the amount of ligand which has bound can be measured. This can be performed with either the wild-type OB-R or the/ ⁇ OB-R of this invention; alternatively the amount of binding to the two alleles can be compared.
  • both the putative ligand and a known ligand are present, and the amount of binding of the putative ligand is compared to the amount of binding to a known ligand.
  • the putative ligand's ability to displace previously bound known ligand may be measured.
  • the assay may be a heterogeneous one, where the OB-R may be bound to a surface, and contacted with putative ligands. Dectection of binding may be by a variety of methods, including labelling, reaction with antibodies, and chomophores.
  • This invention relates to a rat ob receptor which is substantially free from associated rat membrane proteins. It also relates to substantially purified rat ob receptor ("rat OB-R" or "rat OB- receptor") protein.
  • rat OB-R substantially purified rat ob receptor
  • One of the rat OB-Rs of this invention is obtained from a rat which has a wild-type OB-R.
  • Another rat OB-R of this invention is obtained from a rat which has the/ ⁇ mutation.
  • nucleic acids which encode a rat OB receptor.
  • the nucleic acid may be any nucleic acid which can encode a protein, such as genomic DNA, cDNA, or any of the various forms of RNA.
  • the nucleic acid is cDN A.
  • This invention also includes vectors containing a rat OB-R gene, host cells containing the vectors, and methods of making substantially pure rat OB-R protein comprising the steps of introducing a vector comprising a rat OB-R gene into a host cell, and cultivating the host cell under appropriate conditions such that rat OB-R is produced.
  • the rat OB-R so produced may be harvested from the host cells in conventional ways.
  • Yet another aspect of this invention are assays which employ a rat OB-R.
  • various molecules, suspected of being rat OB-R ligands are contacted with a rat OB-R, and their binding is detected.
  • agonists, antagonists, and ligand mimetics may be identified.
  • a further aspect of this invention are the ligands so indentified. The following non-limiting Examples are presented to better illustrate the invention.
  • Tissues were collected from lean and falfa Zucker rats and snap frozen in liquid nitrogen.
  • the tissues collected included: hypothalamus, pituitary, lung, liver, kidney, heart, adrenal glands, smooth muscle, skeletal muscle, and adipose tissue.
  • the tissues were homogenized with a Brinkmann Polytron homogenizer in the presence of guanadinium isothiocyanate.
  • mRNA was prepared from hypothalamus, lung, and kidney according to the instructions provided with the messenger RNA isolation kit (Stratagene, La Jolla, CA).
  • cDNA was prepared from approximately 2 ⁇ g of mRNA with the messenger RNA isolation kit (Stratagene, La Jolla, CA).
  • the first strand cDNA synthesis was primed using 1 ug of oligo(dT)i 2-18 primer and 25 ng of random hexamers per reaction.
  • Second strand cDNA sythesis was performed according to the manufacturer's instructions. The quality of the cDNA was assessed by labeling an aliqout (l/10 tn ) of the second strand reaction with approximately 1 ⁇ Ci of [a-32p]dCTP (3000 Ci/mmol). The labeled products were separated on an agarose gel and detected by autoradiography.
  • the initial portion of the rat OB receptor was obtained by PCR using degenerate primers based on the mouse and human OB- receptor amino acid sequences.
  • the fragments of interest were amplified as long polymerase chain reaction (PCR) products by a modifying the method of Barnes (1994, Proc. Natl. Acad. Sci. 91 :2216- 2220, which is hereby inco ⁇ orated by reference, hi order to obtain the required long PCR fragments, Taq Extender (Stratagene, La Jolla CA.) and the Expand Long Template PCR System (Boehringer Mannheim, Indianapolis, IN) were used in combination.
  • PCR polymerase chain reaction
  • the standard PCR reaction mix in a final volume of 20 ⁇ l, contained 5 ng of template (lean rat cDNA), 100 ng of primers, 500 ⁇ M dNTPs, 1 X Buffer 3 from the Expand kit, 0.1 ⁇ l each of Taq Polymerase and Taq Expander. Reactants were assembled in thin walled reaction tubes.
  • the amplification protocol wasl cycle of 92°C for 30 sec, followed by 32 cycles at 92°C for 30 sec, 45°C for 1 min. and 68°C for 3 min. using a Perkin-Elmer (Norwalk, CT) 9600 Thermal Cycler.
  • This strategy produced a series of PCR products with the largest being approximately 2.2 Kbp amplified from primers ROBR 2 and ROBR 8. These products were subcloned for DNA sequence analysis as described below.
  • PCR products of the appropriate size were prepared for subcloning by separation on an agarose gel, excising the band, and extracting the DNA using Prep-A-Gene (BioRad, Richmond, CA). PCR products were ligated into pCRTMII (Invitrogen, San Diego, CA) according to the instructions provided by the manufacturer. The ligation was transformed into INVaF cells and plated on Luria-Bertani plates containing 100 ⁇ g/ml ampicillin and X-Gal (32 ⁇ l of 50 mg/ml X- Gal (Promega, Madison, WI). White colonies were picked and grown overnight in Luria -Bertani broth plus 100 ⁇ g/ml ampicillin. Plasmid DNAs were prepared using the Wizard miniprep kit (Promega, Madison, WI). Inserts were analyzed by digesting the plasmid DNA with EcoRI and separating the restriction endonulease digestion products on an agarose gel.
  • Plasmid DNA was prepared for DNA sequencing by ethanol precipitation and resuspending in water to achieve a final DNA concentration of 100 ⁇ g/ml.
  • DNA sequence analysis was performed using the ABI PRISMTM dye terminator cycle sequencing ready reaction kit with AmpliTaq DNA polymerase, FS. The initial DNA sequence analysis was performed with Ml 3 forward and reverse primers, subsequently primers based on the rat OB-R sequence were utilized. Following amplification in a Perkin-Elmer 9600, the extension products were purified and analyzed on an ABI PRISM 377 automated sequencer (Perkin Elmer, Norwalk, CT). DNA sequence data was analyzed with the Sequencher program. Due to the unknown genotype of the lean Zucker rat for the/ ⁇ allele, either (+/+ or +/fa) the DNA sequence of multiple subclones of each fragment was analyzed to determine the cDN A sequence of the lean rat OB-R.
  • ROBR 2-8 PCR fragment, rat specific primers ROBR 10 (5'-CTG CAC TTA ACC TGG CCT ATC-3') and ROBR 17 (5'-GGC CAG AAC TGT AAC AGT GTG-3') were synthesized.
  • PCR products were amplified from rat lean hypothalamus, lean lung, falfa hypothalamus and falfa kidney cDNAs.
  • the PCR conditions used for this reaction were a PCR reaction mix with a total volume of 50 ⁇ l containing 5 ng of template (various rat cDNAs mentioned above), 200 ng of primers.
  • Reactants were assembled in thin walled reaction tubes for the Perkin Elmer 9600 Thermal cycler.
  • the amplification protocol was 1 cycle of 92°C for 30 sec, followed by 32 cycles at 92°C for 30 sec, 45°C for 1 min. and 68°C for 4 min. using a Perkin Elmer 9600 Thermal Cycler.
  • Products were then purified, removing all nucleotides and primers, using the QLAquick PCR purification kit according to the manufacturer's specified protocols and resuspended in 30 ⁇ l of water.
  • the second PCR step was then performed using the first PCR reaction as the template and a nested rat specific primer paired with the original 3' primer as outlined above.
  • the reaction conditions were a 50 ⁇ l reaction containing 5 ⁇ l of template (from the purified PCR product), 200 ng of primers, 500 ⁇ M dNTPs, 1 X Buffer 3 from the Expand kit, 0.25 ⁇ l each of Taq Polymerase and Taq Expander. Reactants were assembled in thin walled reaction tubes for the Perkin Elmer 9600 Thermal cycler.
  • the amplification protocol was 1 cycle of 92°C for 30 sec, followed by 25 cycles at 92°C for 30 sec, 45°C for 1 min. and 68°C for 4 min. using a Perkin Elmer 9600 Thermal Cycler.
  • the largest fragment that was generated using the strategy was a fragment produced from ROBR 16 and HOBR I R that was approximately 1500 bp in length.
  • the mouse 3' UTR which presumably encodes a smaller isoform generated by alternative splicing, produced a fragment that was about 650 bp long.
  • the 5' end of the rat OB receptor was obtained by using semi-nested PCR in a manner analogous to that described above for the 3' end.
  • the rat specific primers are the 3' primers that were combined with primers from the 5' UTRs of the human OB-receptor.
  • the primers utilized were HOBR I F (5'-CTT ATG CTG GGA TGT GCC-3 ) and HOBR 1 F-2 (5 -TCG TGG CAT TAT CCT TCA G-3 ) paired with either ROBR 1 1 (5'-GAT AGG CCA GGT TAA GTG CAG-3 ) or ROBR 12 (5'-GAG TGC GGA GCA GTT TTG AC-3).
  • the largest product, HOBR 1F-2 and ROBR 1 1 yielded a 500 bp fragment that covers the region and induces an initiator methionine codon.
  • PCR fragments obtained from falfa cDNA were prepared for DNA sequence analysis by separating the PCR products on an agarose gel, excising the band of interest, and extracting the DNA using Prep-A-Gene (BioRad). Sequencing results of the PCR product generated from falfa hypothalamic cDNA identified a single nucleotide change relative to the lean cDNA sequence. An A to C transversion at bp 880 results in an amino acid change of glutamine to proline at amino acid residue 268. The A to C change in the sequence introduces a Mspl restriction endonuclease site (CCGG) into the sequence.
  • CCGG Mspl restriction endonuclease site
  • falfa cDNAs contained an additional Msp I site identified during the sequencing of ROBR 10/17 and generated products of 747, 1027, and 289. Thus, every tissue examined in e falfa rat was homozygous for the A to C mutation at nucleotide 880.
  • Genomic DNA was prepared from a 2 cm portion of the tail from ten lean and ten falfa Zucker rats and 2 lean and 5 falfa ZDF rats.
  • the tissue was digested overnight at 55°C using 0.3 ⁇ g of Proteinase K in 0.7 ml buffer containing 50 mM Tris, pH 8.0, 100 mM EDTA, and 0.5% SDS.
  • the DNA was extracted two times with phenol/chloroform and one time with chloroform.
  • the DNA was precipitated by adding NaCl to achieve a concentration of 0.3M and then adding an equal volume of 100% ethanol.
  • the DNA was transferred to a 70% wash and then resuspended in 10 mM Tris, 1 mM EDTA.
  • Genomic DNA obtained as outlined above from various sources, was diluted in water to a final concentration of approximately 100 ng/ul.
  • the reaction conditions were a 20 ⁇ l reaction containing I ⁇ l of genomic DNA template, 100 ng of primers, 500 ⁇ M dNTPs, 1 X Buffer 3 from the Expand kit, 0.25 ⁇ l each of Taq Polymerase and Taq Expander. Reactants were assembled in Perkin Elmer 0.5 ml thin walled reaction tubes.
  • the amplification protocol for a Perkin Elmer 480 Thermal Cycler was 32 cycles of 92°C for 30 sec, 54°C for 1 min. and 68°C for 5 min.
  • the products were then analyzed on a 1 % agarose gel.
  • the PCR products contained an endogenous Msp I site that cleaves the fragment somewhere in the intron and produces a 700 bp fragment.
  • the Msp I restriction endonuclease digestion of the 1800 bp ROBR 27/28 PCR product from a homozygous lean rat yields two fragments of 1 100 bp and the endogenous 700 bp fragment.
  • Msp I digestion of PCR products from ⁇ falfa ROBR 27/28 PCR amplification which contains the A to C mutation, introduces an additional Msp I site that cleaves the 1 100 bp band to produce a 950 bp and a small fragment of 130 bp.

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Abstract

Gène récepteur de l'obésité du rat (ob) isolé et cloné. Deux différents allèles ont été identifiés: le type sauvage et l'allèle fa, qui diffère du précédent seulement par une paire de base. Le changement d'une paire de base introduit toutefois un site de restriction MspI dans la séquence ADN et entraîne également une modification d'acide aminé. L'invention porte également sur de nouveaux récepteurs, sur des vecteurs renfermant l'acide nucléique codant ces récepteurs, sur des cellules hôtes transformées par ce gène et sur des dosages mettant en oeuvre ce gène ou cette protéine pour identifier de nouveaux ligands.
PCT/US1997/002397 1996-02-22 1997-02-18 Recepteurs de l'obesite du rat et nucleotides codant ces recepteurs WO1997031015A1 (fr)

Priority Applications (2)

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JP53025497A JP2002515739A (ja) 1996-02-22 1997-02-18 ラットｏｂレセプター及びそれらをコードするヌクレオチド
EP97905980A EP0922052A4 (fr) 1996-02-22 1997-02-18 Recepteurs de l'obesite du rat et nucleotides codant ces recepteurs

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
US9040596P 1996-02-22 1996-02-22
US60/090,405 1996-02-22
US1396996P 1996-03-22 1996-03-22
US60/013,969 1996-03-22
GBGB9608473.6A GB9608473D0 (en) 1996-04-25 1996-04-25 Rat OB receptors and nucleotides encoding them
GB9608473.6 1996-04-25

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0900282A1 (fr) * 1996-05-06 1999-03-10 Merck & Co., Inc. ISOFORMES DE RECEPTEUR DU GENE $i(OB) ET ACIDES NUCLEIQUES LES CODANT
US5935810A (en) * 1994-08-17 1999-08-10 The Rockefeller University Mammalian ob polypeptides capable of modulating body weight, corresponding nucleic acids, and diagnostic and therapeutic uses thereof
US6258944B1 (en) * 1996-05-06 2001-07-10 Merck & Co., Inc. OB receptor isoforms and nucleic acids encoding them
US7063958B1 (en) 1996-01-16 2006-06-20 The Rockefeller University Nucleic acids db, the receptor for leptin
US7084252B1 (en) 1996-01-16 2006-08-01 The Rockefeller University DB, the receptor for leptin
US7148004B1 (en) 1997-01-16 2006-12-12 The Rockefeller University Oligonucleotides of the OB-R isoforms and methods of diagnosing body weight
US7619079B2 (en) 1996-02-14 2009-11-17 The Rockefeller University Db, the receptor for leptin, nucleic acids encoding the receptor, and uses thereof

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 26 April 1995, Volume 209, Number 3, MURAKAMI et al., "Cloning of Rat OBESE cDNA and its Expression in Obese Rats", pages 944-952. *
CELL, 29 December 1995, Volume 83, TARTAGLIA et al., "Identification and Expression Cloning of a Leptin Receptor, OB-R", pages 1263-1271. *
NATURE GENETICS, May 1996, Volume 13, No. 1, PHILLIPS et al., "Leptin Receptor Missense Mutation in the Fatty Zucker Rat", pages 18-19. *
See also references of EP0922052A4 *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5935810A (en) * 1994-08-17 1999-08-10 The Rockefeller University Mammalian ob polypeptides capable of modulating body weight, corresponding nucleic acids, and diagnostic and therapeutic uses thereof
US7521258B2 (en) 1994-08-17 2009-04-21 The Rockefeller University Methods of detecting, measuring, and evaluating modulators of body weight in biological samples, and diagnostic, monitoring, and therapeutic uses thereof
US7063958B1 (en) 1996-01-16 2006-06-20 The Rockefeller University Nucleic acids db, the receptor for leptin
US7084252B1 (en) 1996-01-16 2006-08-01 The Rockefeller University DB, the receptor for leptin
US7612171B2 (en) 1996-01-16 2009-11-03 The Rockefeller University DB, the receptor for leptin, nucleic acids encoding the receptor, and uses thereof
US7812137B2 (en) 1996-01-16 2010-10-12 The Rockefeller University Db, the receptor for leptin, nucleic acids encoding the receptor, and uses thereof
US7619079B2 (en) 1996-02-14 2009-11-17 The Rockefeller University Db, the receptor for leptin, nucleic acids encoding the receptor, and uses thereof
EP0900282A1 (fr) * 1996-05-06 1999-03-10 Merck & Co., Inc. ISOFORMES DE RECEPTEUR DU GENE $i(OB) ET ACIDES NUCLEIQUES LES CODANT
EP0900282A4 (fr) * 1996-05-06 2001-04-11 Merck & Co Inc Isoformes de recepteur du gene ob et acides nucleiques les codant
US6258944B1 (en) * 1996-05-06 2001-07-10 Merck & Co., Inc. OB receptor isoforms and nucleic acids encoding them
US7148004B1 (en) 1997-01-16 2006-12-12 The Rockefeller University Oligonucleotides of the OB-R isoforms and methods of diagnosing body weight

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JP2002515739A (ja) 2002-05-28
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EP0922052A1 (fr) 1999-06-16

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