CN115053861A - Construction method and application of animal model of schizophrenia based on immune activation - Google Patents
Construction method and application of animal model of schizophrenia based on immune activation Download PDFInfo
- Publication number
- CN115053861A CN115053861A CN202210763150.2A CN202210763150A CN115053861A CN 115053861 A CN115053861 A CN 115053861A CN 202210763150 A CN202210763150 A CN 202210763150A CN 115053861 A CN115053861 A CN 115053861A
- Authority
- CN
- China
- Prior art keywords
- schizophrenia
- immune activation
- animal
- animal model
- female
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 201000000980 schizophrenia Diseases 0.000 title claims abstract description 38
- 238000010171 animal model Methods 0.000 title claims abstract description 36
- 230000005934 immune activation Effects 0.000 title claims abstract description 28
- 238000010276 construction Methods 0.000 title claims abstract description 10
- 230000008774 maternal effect Effects 0.000 claims abstract description 19
- 230000035935 pregnancy Effects 0.000 claims abstract description 16
- 239000002158 endotoxin Substances 0.000 claims abstract description 15
- 229920006008 lipopolysaccharide Polymers 0.000 claims abstract description 15
- 241001465754 Metazoa Species 0.000 claims abstract description 13
- 239000003814 drug Substances 0.000 claims abstract description 7
- 229940079593 drug Drugs 0.000 claims abstract description 5
- 238000000034 method Methods 0.000 claims description 19
- 238000012216 screening Methods 0.000 claims description 4
- 230000037396 body weight Effects 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 abstract description 50
- 241000699670 Mus sp. Species 0.000 abstract description 35
- 238000002474 experimental method Methods 0.000 abstract description 20
- 230000003542 behavioural effect Effects 0.000 abstract description 10
- 210000004556 brain Anatomy 0.000 abstract description 4
- 239000013614 RNA sample Substances 0.000 abstract description 3
- 238000012163 sequencing technique Methods 0.000 abstract description 3
- 238000010201 enrichment analysis Methods 0.000 abstract description 2
- 230000007246 mechanism Effects 0.000 abstract description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 15
- 238000012360 testing method Methods 0.000 description 11
- 201000010099 disease Diseases 0.000 description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 8
- 210000005013 brain tissue Anatomy 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 230000037361 pathway Effects 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 238000003559 RNA-seq method Methods 0.000 description 4
- 230000007613 environmental effect Effects 0.000 description 4
- 210000000225 synapse Anatomy 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 208000019901 Anxiety disease Diseases 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 230000006870 function Effects 0.000 description 3
- 230000002068 genetic effect Effects 0.000 description 3
- 208000026350 Inborn Genetic disease Diseases 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 230000036506 anxiety Effects 0.000 description 2
- 230000007529 anxiety like behavior Effects 0.000 description 2
- 230000006399 behavior Effects 0.000 description 2
- 230000031018 biological processes and functions Effects 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 230000008451 emotion Effects 0.000 description 2
- 208000016361 genetic disease Diseases 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 230000013011 mating Effects 0.000 description 2
- 101150028074 2 gene Proteins 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 208000031404 Chromosome Aberrations Diseases 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 206010013654 Drug abuse Diseases 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 238000000729 Fisher's exact test Methods 0.000 description 1
- YQEZLKZALYSWHR-UHFFFAOYSA-N Ketamine Chemical compound C=1C=CC=C(Cl)C=1C1(NC)CCCCC1=O YQEZLKZALYSWHR-UHFFFAOYSA-N 0.000 description 1
- 206010057249 Phagocytosis Diseases 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000030741 antigen processing and presentation Effects 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 238000009227 behaviour therapy Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 210000003850 cellular structure Anatomy 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000035606 childbirth Effects 0.000 description 1
- 231100000005 chromosome aberration Toxicity 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000010219 correlation analysis Methods 0.000 description 1
- 210000004292 cytoskeleton Anatomy 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000008995 epigenetic change Effects 0.000 description 1
- 230000002964 excitative effect Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000033209 immune effector process Effects 0.000 description 1
- 230000008102 immune modulation Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 229960003299 ketamine Drugs 0.000 description 1
- 230000008183 lymphocyte mediated immunity Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 238000012346 open field test Methods 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000008447 perception Effects 0.000 description 1
- 230000008782 phagocytosis Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 230000003234 polygenic effect Effects 0.000 description 1
- 230000001242 postsynaptic effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000003518 presynaptic effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 208000020016 psychiatric disease Diseases 0.000 description 1
- 230000000384 rearing effect Effects 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 208000011117 substance-related disease Diseases 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000009182 swimming Effects 0.000 description 1
- 230000005062 synaptic transmission Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/02—Breeding vertebrates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
- A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2207/00—Modified animals
- A01K2207/20—Animals treated with compounds which are neither proteins nor nucleic acids
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
- A01K2267/035—Animal model for multifactorial diseases
- A01K2267/0356—Animal model for processes and diseases of the central nervous system, e.g. stress, learning, schizophrenia, pain, epilepsy
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Endocrinology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Diabetes (AREA)
- Animal Husbandry (AREA)
- Gastroenterology & Hepatology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Rheumatology (AREA)
- Toxicology (AREA)
- Urology & Nephrology (AREA)
- Epidemiology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention belongs to the field of biological medicine, and particularly relates to a construction method and application of an immune activation-based schizophrenia animal model, wherein the construction method of the animal model comprises the following steps: lipopolysaccharide is utilized to treat pregnant female experimental animals to induce maternal immune activation, and first generation animals bred by the pregnant female experimental animals are the animal models of schizophrenia based on immune activation. The invention carries out behavioral experiments on maternal immune activation female offspring mice at different periods, prepares whole brain RNA samples for transcriptome sequencing, carries out enrichment analysis on differential expression genes and schizophrenia related genes, finds the relation between maternal immune activation and schizophrenia, treats experimental animals in gestation period by lipopolysaccharide based on the mechanism, takes the maternal immune activation female offspring mice as animal models of schizophrenia, and has important significance for researching schizophrenia induced by environment or drugs.
Description
Technical Field
The invention belongs to the field of biological medicines, and particularly relates to a construction method and application of an immune activation-based schizophrenia animal model.
Background
Schizophrenia involves various dysfunctions such as perception, memory, thinking, emotion, language, social intercourse, emotion, will behavior and the like to different degrees, seriously affects the interplay, learning, work, life, self-care ability and the like of patients, and brings huge economic and life pressure to society and families. At present, the incidence rate of schizophrenia is about 1 percent, and the disease is in a trend of increasing year by year, the disease is a polygenic complex genetic disease and is determined by genetic factors and environmental factors, and the genetic factors mainly comprise chromosome aberration, point mutation and epigenetic change; environmental factors mainly include stress, infection, environmental pollution, drug abuse, and the like. In recent years, a plurality of researches show that proinflammatory cytokines can be detected at higher level in blood of mental disease patients, and with the deep research on schizophrenia, animal models become important means and tools for researchers at home and abroad to research pathogenesis and clinical characteristics of the disease. Therefore, establishing a novel schizophrenia model is an important means for researching the disease.
At present, the method for constructing the animal model of schizophrenia generally comprises the steps of directly injecting medicaments (such as ketamine, PCP, MIA-801 and the like) into a mouse body, and then performing behavioral tests on the mouse, the method is single, and related researches show that the disease is a polygene complex genetic disease and is determined by genetic factors and environmental factors.
Disclosure of Invention
Aiming at the problems in the prior art, the invention aims to provide a construction method and application of an immune activation-based schizophrenia animal model.
Based on the above purpose, the technical scheme adopted by the invention is as follows:
in a first aspect, the invention provides a method for constructing an immune-activated schizophrenia-based animal model, which comprises the following steps: lipopolysaccharide is used for treating experimental animals in gestation period to induce maternal immune activation, and the first generation animals bred by the immune-activated experimental animals in gestation period are the animal models of schizophrenia based on immune activation.
Preferably, the method for treating the experimental female animals in gestation period by using lipopolysaccharide comprises the following steps: on the 16 th day of gestation of the experimental animals, the lipopolysaccharide solution was intraperitoneally injected at a dose of 150. mu.g/kg, according to the animal body weight.
Preferably, the concentration of the lipopolysaccharide solution is 1 mg/ml.
Preferably, the first generation animal is a female animal.
Preferably, the experimental animal is a mouse.
In a second aspect, the invention provides an application of an immune-activated schizophrenia-based animal model in preparation or screening of drugs for treating schizophrenia, wherein the immune-activated schizophrenia-based animal model is constructed by the method.
Compared with the prior art, the invention has the following beneficial effects:
the invention firstly carries out behavioral experiments on maternal immune activation female offspring mice at different periods, prepares a whole brain RNA sample for transcriptome sequencing, carries out enrichment analysis on differentially expressed genes and schizophrenia related genes, finds the relation between maternal immune activation and schizophrenia, treats gestational experimental animals by lipopolysaccharide based on the mechanism, takes the maternal immune activation female offspring mice as animal models of schizophrenia, and has important significance for researching schizophrenia induced by environment or drugs.
Drawings
FIG. 1 is a plan view of a maternal immune activation experiment;
FIG. 2 is a plan view of a maternal immune activation experiment;
FIG. 3 shows the results of an elevated plus maze experiment;
FIG. 4 is a heat map of differentially expressed genes in brain tissue at four time points during the development of maternal immune-activated female offspring;
FIG. 5 shows the enrichment of genes differentially expressed from schizophrenia related genes;
FIG. 6 shows the immune-related pathways and biological functions of the enrichment of differentially expressed genes in brain tissue of four time points of maternal immune activation of female offspring.
Detailed Description
To better illustrate the objects, aspects and advantages of the present invention, the present invention will be further described with reference to specific examples. It will be understood by those skilled in the art that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The test methods used in the examples are all conventional methods unless otherwise specified; the materials, reagents and the like used are commercially available unless otherwise specified.
The female mice used in the examples were provided by the southern medical university animal center, and the rearing conditions were in accordance with the animal room requirements, the temperature was 21-24 ℃, the relative humidity was 50-70%, and the mice were reared in autoclaved water, and had free access to food and water. The feed cage bedding was autoclaved and sterilized and was changed twice a week. All the operations meet the requirements related to southern medical university laboratory animal ethical Specification.
Example 1 construction of animal models and phenotypic validation
Selecting wild SPF female mice and male mice (8 weeks old) to mate, judging the pregnancy time of the mice according to vaginal emboli of the mice, selecting 20 pregnant female mice, randomly dividing the pregnant female mice into two groups with equal quantity, wherein one group is subjected to intraperitoneal injection of LPS solution according to the weight of the female mice and the dosage of 150ug/kg on the 16 th day of pregnancy of the female mice so as to simulate bacterial infection and induce the immune activation of a parent, and the LPS solution is prepared from normal saline with the concentration of 1mg/ml and can be diluted and injected according to the weight condition of animals in actual use.
Another group of pregnant females injected an equal volume of normal saline on day 16 of gestation.
After the injection is finished, the mice are placed in a mouse cage for continuous breeding, the mice are waited for production, after offspring mice are weaned, female offspring are picked out for breeding, and the female offspring are grouped for experiment.
The first generation female mice after birth were subjected to behavioural tests at different developmental time points (puberty, 7 days after delivery and 21 days after delivery), the technical route of which is shown in fig. 1, such as a bead burying test, an open field test, an elevated plus maze test, a sugar water consumption test, a forced swimming test and a water maze test, and the anxiety-like phenotype of the mice at different time points is found to be obvious.
Behavioral experiments were performed at week 8 (Preg0) including 8 female mice in the experimental group (L _ Preg0) and 8 female mice in the control group (C _ Preg 0); the experimental group female mice and the control group female mice are first generation female mice delivered by immune activation and physiological saline treated pregnant mice respectively; the 8 th week after birth of the first generation female mouse is adolescence.
Mating with male mice in a cage at the 8 th week of the growth of the first generation female mice, and performing a behavioral experiment including an experimental group (L _ AD7) and a control group (C _ AD7) 7 days after birth (AD 7);
mating with male mice in cages at week 8, and performing behavioral experiments on day 21 postpartum (AD21), including an experimental group (L _ AD21) and a control group (C _ AD 21);
behavioral experiments were performed on the 84 th day of birth (P84, corresponding in time to AD7) on mice mated with cages, including L _ P84 and control C _ P84.
And extracting brain tissue RNA from female offspring subjected to LPS-induced maternal immune activation and a control group at different time points (Preg0, Preg19, AD2 and AD21), and sending the brain tissue RNA to a company for RNA-seq, as shown in figure 2, because the body of a mouse is relatively bloated in 19 days of gestation and relatively weak in 2 days of childbirth, the experimental study on behaviors is inconvenient. We wanted to study brain transcriptomics changes during pregnancy and just after delivery of mice, so we performed behavioural experiments 7 days post-partum (AD7), RNA-seq 2 days post-partum (AD2), and increased the time point of 19 days gestation (Preg19) of female offspring in samples of RNA-seq.
In the behavioural experiment, take the overhead cross maze experiment of mouse as an example, before the experiment begins, move the mouse to the experiment room from the animal room to adaptation environmental time needs to reach 1 hour and more, need to guarantee that the experimental apparatus is clean and free from extraneous odour, enclose overhead cross maze device with the curtain and avoid external interference, erect the camera, record the position of mouse, data such as dwell time with video tracking software (Biobserve, Fort Lee, NJ). During formal testing, a mouse to be tested is placed at the intersection of the open arm and the closed arm of the elevated plus maze, the test time is 5 minutes facing the open arm, and after the test of the mouse is finished each time, 75% alcohol is sprayed on the elevated plus maze device to clean the excrement of the mouse. According to the time of the mouse staying at the open arm and the closed arm recorded by the software, the percentage of the time of staying at the open arm is counted, and the lower the proportion is, the more anxiety the mouse is.
The results of the experiments in each group are shown in fig. 3, fig. 3A shows that the proportion of the open-arm residence time of the elevated cross maze experiment in the group C _ Preg0 and the proportion of the open-arm residence time of the elevated cross maze experiment in the group L _ Preg0 in the group L _ Preg0 are reduced, and the difference is statistically significant (n is 8, p is less than 0.05). It was shown that maternal immune-activated female offspring showed anxiety-like behavior at the same time period of adolescence 8 weeks (L _ Preg0) compared to control mice (C _ Preg 0).
Fig. 3B shows the results of the proportion of open-arm residence time to total time in the elevated maze experiments in C _ P84 group and L _ P84 group, and the proportion of open-arm residence time in L _ P84 group is decreased, and the difference is statistically significant (n is 8, P is less than 0.05). Maternal immune-activated female offspring were shown to exhibit anxiety-like behavior at day 84 of birth in adulthood (L _ P84) compared to control mice in the same period (C _ P84).
Fig. 3C shows the proportion of the open-arm residence time in the elevated plus maze experiments of the C _ AD7 group and the L _ AD7 group, and the C _ P84 group and the L _ P84 group in the total time, and the percentage of the open-arm residence time in the L _ AD7 group is reduced less than that in the C _ AD7 group, and the difference is statistically significant (n is 8, P is less than 0.05), and the difference in the L _ AD7 group is not statistically significant (n is 8, P is more than 0.05) than that in the L _ P84 group. Fig. 3D shows the proportion of open-arm residence time in the elevated plus maze test in groups C _ AD21 and L _ AD21 to the total time, and the percentage of open-arm residence time in group L _ AD7 was reduced less than that in group C _ AD7, with statistical differences (n is 8, p is < 0.05). Female offspring mice showing maternal immune activation showed anxiety at this time point of 7 days post partum, but the process of pregnancy delivery did not constitute a "second hit".
Example 2 Gene correlation analysis of animal models
Female offspring activated by LPS-induced maternal immunity and control group extracted brain tissue RNA at different time points (Preg0, Preg19, AD2, AD21) and sent to the company for RNA-seq.
Mouse whole brain RNA samples were prepared for transcriptome sequencing. For the identification of the model at the gene level, the differential expression of the gene was calculated at each time point using DEseq2(v1.24.0) and edgeR (v2.36.8) (based on the R language platform), and the gene with false positive discovery rate (FDR) less than 0.05 in both methods was taken as the differentially expressed gene.
The two methods of the edgeR method and the DESeq2 are used for screening out a wien graph of the differential expression genes, and the fact that the intersection number of the differential expression genes screened by the two methods is higher is shown. For example, in the Preg0 group, 1932 differentially expressed genes were obtained by the edgeR method, 2057 differentially expressed genes were obtained by the DESeq2 method, 1875 genes were intersected among the differentially expressed genes obtained by the two methods, and the occupation ratio was up to 90%, and similar results were also found at other time points, indicating the reliability of downstream analysis using the intersection of the results obtained by the two methods as the differentially expressed genes. In addition, the expression heatmap of differentially expressed genes (fig. 4) suggested that the differentially expressed genes were expressed substantially uniformly within the group, but that there was a significant difference between the experimental group and the control group, which also indicates that the differentially expressed genes were highly reliable.
1449 schizophrenia candidate genes are collected from an SZGR database (https:// bioinfo. uth. edu/SZGR /), the proportion of the schizophrenia candidate genes in the background genes is used as expected disease gene proportion (expected) with the proportion of the expression genes at each time point as background genes, the proportion of the schizophrenia candidate genes in the background genes in the four time point groups is used as disease gene proportion, and a unilateral Fisher's exact test (based on an R language platform) is carried out by using a fisher' test function (based on the R language platform) to calculate the enrichment condition of the different expression genes of the above candidate genes in the different time point groups. The standard error was calculated by randomly repeating the extraction of 80% of the genes among the differentially expressed genes 1000 times to calculate the proportion of disease genes, and the results are shown in fig. 5, which indicates that the differentially expressed genes of Preg0 group and Preg19 group are enriched in genes related to schizophrenia.
Based on the R language platform, the clusterProfiler package (v3.12.0) was used to analyze the function of the differentially expressed genes and the enrichment pathway. Further explores the relationship between maternal immune activation and schizophrenia. The Preg0 time point differentially expressed gene enrichment pathway, cellular components, and cellular functions were found to be involved in both immune and synaptic transmission, as shown in fig. 6, fig. 6A is a KEGG pathway enriched for four time points differentially expressed genes. The KEGG signal pathways enriched by the differential expression genes in each time point group and related pathways such as virus infection, tuberculosis, cell phagocytosis and the like indicate that the differential expression genes mainly participate in the immune response of brain tissues. Fig. 6B shows GO cell fractions enriched for differentially expressed genes at four time points, where only Preg0 group differentially expressed genes enriched for synapse-associated cell fractions such as presynaptic membrane, neuronal synapses, excitatory synapses, and postsynaptic specialization, while other time points (Preg19, AD2, AD21) did not enrich for synapse-associated functions. The differentially expressed genes at four time points are all enriched in related cell components such as cell membranes, cytoskeletons and the like. Figure 6C GO biological process of differential expression gene enrichment at four time points. The biological processes involved include viral feedback, antigen processing, lymphocyte-mediated immunity, immune responses, and modulation of immune effector processes, most of which are associated with immunity. The results further prove that the model constructed by the invention is an animal model of schizophrenia from a gene level.
In conclusion, the schizophrenia animal model is constructed by the immune-activated parent for the first time, the behavioural screening is carried out on the female offspring mice immune-activated by the parent, and researches show that the brain tissue differential expression gene of the offspring female mice is related to the enrichment condition of the schizophrenia-related gene, so that the model construction method is effective and feasible.
Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
Claims (6)
1. A construction method of an animal model of schizophrenia based on immune activation is characterized by comprising the following steps: lipopolysaccharide is used for treating experimental animals in gestation period to induce maternal immune activation, and the first generation animals bred by the immune-activated experimental animals in gestation period are the animal models of schizophrenia based on immune activation.
2. The construction method according to claim 1, wherein the method for treating the experimental animal in gestation period by using lipopolysaccharide comprises: on the 16 th day of gestation of the experimental animals, the lipopolysaccharide solution was intraperitoneally injected at a dose of 150. mu.g/kg, according to the animal body weight.
3. The method of claim 2, wherein the lipopolysaccharide solution has a concentration of 1 mg/ml.
4. The method of claim 1, wherein the first generation animal is a female animal.
5. The method of claim 1, wherein the experimental animal is a mouse.
6. The application of an immune activation-based schizophrenia animal model in preparing or screening drugs for treating schizophrenia is characterized in that the immune activation-based schizophrenia animal model is constructed by the method of any one of claims 1-5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210763150.2A CN115053861A (en) | 2022-06-30 | 2022-06-30 | Construction method and application of animal model of schizophrenia based on immune activation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210763150.2A CN115053861A (en) | 2022-06-30 | 2022-06-30 | Construction method and application of animal model of schizophrenia based on immune activation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN115053861A true CN115053861A (en) | 2022-09-16 |
Family
ID=83204837
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210763150.2A Pending CN115053861A (en) | 2022-06-30 | 2022-06-30 | Construction method and application of animal model of schizophrenia based on immune activation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115053861A (en) |
Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007131041A2 (en) * | 2006-05-02 | 2007-11-15 | Corcept Therapeutics, Inc. | The use of a glucocorticoid receptor ii antagonist to treat depression in patients taking il-2 |
| US20080176240A1 (en) * | 2007-01-19 | 2008-07-24 | Smithkline Beecham Corporation | Genes associated with schizophrenia identified using a whole genome scan |
| US20110201565A1 (en) * | 2008-10-30 | 2011-08-18 | University Of South Florida | Luteolin and diosmin/diosmetin as novel stat3 inhibitors for treating autism |
| CN105164264A (en) * | 2012-12-12 | 2015-12-16 | 布罗德研究所有限公司 | Delivery, engineering and optimization of systems, methods and compositions for sequence manipulation and therapeutic applications |
| CN107760713A (en) * | 2016-08-23 | 2018-03-06 | 中国科学院上海药物研究所 | A kind of method for building up of non-human mammal neuropsychiatric disease animal model and application thereof |
| CN107801692A (en) * | 2017-10-23 | 2018-03-16 | 昆明医科大学 | A kind of ketamine induction animal model of schizophrenia and its Mechanism Study |
| TW201831175A (en) * | 2017-02-27 | 2018-09-01 | 中原大學 | Uses of n-acetylcysteine for the prophylaxis of preterm birth, still birth, fetal death, and/or fetal brain injury |
| CN110583572A (en) * | 2019-09-18 | 2019-12-20 | 南方医科大学南方医院 | Construction method for simultaneously constructing memory impairment model, anxiety model and strong aggressive mouse model |
| CN110693894A (en) * | 2019-11-18 | 2020-01-17 | 贵州中医药大学 | Application of neuroprotective effect of dipsacoside VI in treatment of depression |
| JP2020015710A (en) * | 2018-07-27 | 2020-01-30 | 永喜 廣澤 | Method for preventing, improving and treating intractable diseases by pathogenic microorganisms and the like, lps, and quorum sensing |
| CN114128672A (en) * | 2021-11-21 | 2022-03-04 | 河南省儿童医院郑州儿童医院 | Construction method of autism rat model |
| US20220072101A1 (en) * | 2019-01-11 | 2022-03-10 | President and Fellows of Haravrd College | Harnessing inflammation to treat neurodevelopmental disorders |
-
2022
- 2022-06-30 CN CN202210763150.2A patent/CN115053861A/en active Pending
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007131041A2 (en) * | 2006-05-02 | 2007-11-15 | Corcept Therapeutics, Inc. | The use of a glucocorticoid receptor ii antagonist to treat depression in patients taking il-2 |
| US20080176240A1 (en) * | 2007-01-19 | 2008-07-24 | Smithkline Beecham Corporation | Genes associated with schizophrenia identified using a whole genome scan |
| US20110201565A1 (en) * | 2008-10-30 | 2011-08-18 | University Of South Florida | Luteolin and diosmin/diosmetin as novel stat3 inhibitors for treating autism |
| CN105164264A (en) * | 2012-12-12 | 2015-12-16 | 布罗德研究所有限公司 | Delivery, engineering and optimization of systems, methods and compositions for sequence manipulation and therapeutic applications |
| CN107760713A (en) * | 2016-08-23 | 2018-03-06 | 中国科学院上海药物研究所 | A kind of method for building up of non-human mammal neuropsychiatric disease animal model and application thereof |
| TW201831175A (en) * | 2017-02-27 | 2018-09-01 | 中原大學 | Uses of n-acetylcysteine for the prophylaxis of preterm birth, still birth, fetal death, and/or fetal brain injury |
| CN107801692A (en) * | 2017-10-23 | 2018-03-16 | 昆明医科大学 | A kind of ketamine induction animal model of schizophrenia and its Mechanism Study |
| JP2020015710A (en) * | 2018-07-27 | 2020-01-30 | 永喜 廣澤 | Method for preventing, improving and treating intractable diseases by pathogenic microorganisms and the like, lps, and quorum sensing |
| US20220072101A1 (en) * | 2019-01-11 | 2022-03-10 | President and Fellows of Haravrd College | Harnessing inflammation to treat neurodevelopmental disorders |
| CN110583572A (en) * | 2019-09-18 | 2019-12-20 | 南方医科大学南方医院 | Construction method for simultaneously constructing memory impairment model, anxiety model and strong aggressive mouse model |
| CN110693894A (en) * | 2019-11-18 | 2020-01-17 | 贵州中医药大学 | Application of neuroprotective effect of dipsacoside VI in treatment of depression |
| CN114128672A (en) * | 2021-11-21 | 2022-03-04 | 河南省儿童医院郑州儿童医院 | Construction method of autism rat model |
Non-Patent Citations (4)
| Title |
|---|
| AMANDA C. KENTNER等: "Environmental enrichment rescues the effects of early life inflammation", 《BRAIN, BEHAVIOR, AND IMMUNITY》 * |
| E.J.CONNORS等: "Environmental enrichment mitigates the sex-specific effects of gestational inflammation on social engagement and the hypothalamic pituitary adrenal axis-feedback system", 《BRAIN, BEHAVIOR, AND IMMUNITY》 * |
| 冯飞等: "双向情感障碍的内表型研究进展", 《汕头大学医学院学报》 * |
| 吕静妍等: "精神分裂症啮齿类动物模型的构建", 《国际精神病学杂志》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Koolhaas et al. | Animal behavior and well-being symposium: Interaction between coping style/personality, stress, and welfare: Relevance for domestic farm animals | |
| Kiani et al. | Ethical considerations regarding animal experimentation | |
| CA2796822C (en) | Measurement and comparison of immune diversity by high-throughput sequencing | |
| Hau | Animal models for human diseases: an overview | |
| Uekita et al. | Hippocampus lesions induced deficits in social and spatial recognition in Octodon degus | |
| Holmes | Sibling recognition in thirteen-lined ground squirrels: effects of genetic relatedness, rearing association, and olfaction | |
| Li et al. | Maternal immune activation alters adult behavior, gut microbiome and juvenile brain oscillations in ferrets | |
| Samineni et al. | Cellular, circuit and transcriptional framework for modulation of itch in the central amygdala | |
| CN114038574A (en) | Drug relocation system and method based on heterogeneous association network deep learning | |
| Patterson | Infectious behavior: brain-immune connections in autism, schizophrenia, and depression | |
| CN112245596A (en) | Method for screening drug effect substances based on zebra fish inflammatory bowel disease model | |
| Santana-Coelho et al. | Advancing autism research from mice to marmosets: Behavioral development of offspring following prenatal maternal immune activation | |
| CN109276711B (en) | Application of manganese type high-stability superoxide dismutase in improving autism and product | |
| Seelke et al. | Fatherhood alters gene expression within the MPOA | |
| CN115053861A (en) | Construction method and application of animal model of schizophrenia based on immune activation | |
| CN111388684A (en) | Method for evaluating anti-Alzheimer disease efficacy of drug by using transgenic zebra fish | |
| Vasilevskaya et al. | Local experimental intracerebral hemorrhage in rats | |
| Barkan et al. | Neuronal recruitment in adult zebra finch brain during a reproductive cycle | |
| O'Connor et al. | Adolescent environmental enrichment induces social resilience and alters neural gene expression in a selectively bred rodent model with anxious phenotype | |
| CA2964359A1 (en) | Methods for conducting stimulus-response studies with induced pluripotent stem cells derived from perinatal cells or tissues | |
| CN110305949A (en) | The analysis of schizophrenia disease mouse model hippocampus mRNA sequence and kit | |
| Pillay et al. | Mating strategy predicts the occurrence of the Bruce effect in the vlei rat Otomys irroratus | |
| CN110396541A (en) | Schizophrenia disease mouse model hippocampus lncRNA sequencing analysis and kit | |
| Zhang et al. | Integration of genetic and Immune Infiltration insights into Data Mining of multiple sclerosis pathogenesis | |
| CN116179601B (en) | Method for constructing autism disease animal model with social disorder and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20220916 |
|
| RJ01 | Rejection of invention patent application after publication |