Construction And Intervention Of 16P11.2 Microdeletion Autism Spectrum Disorder Mouse Model

Background and objective:Autism spectrum disorder ASD is a group of severe mental and neuro developmental disorders that occur in early childhood,characterised by social communication and interaction impairments,repeated restricted patterns of behaviour,and severe physical disfunction.At present,because the pathogenesis of ASD is not clear,and the prevalence rate is increasing year by year,the pathogenesis and intervention research of ASD have become a hot topic in neuroscience research.16P11.2 bits are slightly missing.Attention deficits and destructive behavioural disorders caused by single copies of 26 genes are one of the most common chromosome aberrations related to ASD.Therefore,the establishment of autism spectral disorder mice with 16P11.2 microdeletion is one of the common models for ASD research at present.At present,the treatment of ASD is mainly to interfere with the patient’s behavior.Changing the patient’s intestinal flora to regulate the behavior of ASD patients is one of the treatment methods.As a weak 4-aminobutyric acid(GABA)A receptor agonist,taurine can improve ASD behavior by affecting GABA signaling and producing neuroactive metabolites.At the same time,neural stem cells and mesenchymal stem cells,as stem cells with self-renewal and multi-differentiation potential,can effectively treat neurological diseases and provide a new way for intervention and treatment of ASD.Method:1.CRISPR/Cas9 system builds a 16P11.2 microdeletion ASD mouse model,and genotypes of mice are identified by PCR and Western blot detection.2.Behavioural test of 16P11.2 microdeletion ASD mice was carried out,and 16P11.2del mice were identified at the animal behaviour level.3.Taurine treats ASD mice and detects the therapeutic effect by behavioural methods.4.Culture 16P11.2del and WT neural stem cells and mesenchymal stem cells.Taurine treats ASD mice and detects the therapeutic effect by behavioural methods.5.Test 16P11.2del and WT neural stem cells and mesenchymal stem cells for cell level identification,including fluorescence staining experiments,fluid staining analysis,proliferation experiments and differentiation experiments,to detect changes in cell development levels in sick mice.Results:1.The results of the open field experiment show that the action time of 16P11.2del mice in the central area is 40% shorter than that of WT mice(P<0.01)2.The number of black marbles embedded in the litter of 2.16P11.2del mice is about 1.5 times the number embedded in WT mice(P<0.001).3.Compared with WT mice,16P11.2del mice have reduced the number of explorations for open arms by 20%(P<0.05).4.16P11.2del mice and WT mice have no significant difference in sniffing time for new and old objects.5.16P11.2del mice have their forefoot off the ground,the number of standing times is 1.25 times that of WT mice(P<0.001),and the time of self-grooming behavior is twice that of WT mice(P<0.001).6.After taurine drug intervention treatment,16P11.2del mice compared with untreated 16P11.2del mice,the action time in the central area was significantly increased,and they were untreated 16P11.2del mice The number of beads buried was significantly reduced to 60% of the original number,and the frequency of standing behavior was significantly reduced to 60% of the original number.7.Compared with the control group,the proliferation ability of NSCs in 16P11.2del mice was reduced by 30%.8.After osteogenic differentiation,the area of calcium nodules in MSCs of16P11.2del mice was reduced by 80% compared with MSCs of wild-type mice,and the degree of osteogenic differentiation was lower.9.The results of adipogenic differentiation showed that the number of lipid droplets in16P11.2del MSCs was 3 times that of WT MSCs.Conclusion:1.Detections of 16P11.2 microdeletion ASD mice commissioned by the company confirmed that the level of gene expression related to this chromosome decreased.Behavioural tests found that there were obvious anxiety behaviours and repeated rigid behaviours compared with the control group mice.It shows the success of 16P11.2microdeletion autism spectrum disorder mice.2.16P11.2 micro-deficient ASD mice were treated by Taurine,and anxiety behaviour and repeated rigid behaviour improved significantly.3.16P11.2 microdeletion inhibits the proliferation ability of NSCs and the osteogenesis ability of BM-MSCs,and enhances the lipid differentiation ability of BM-MSCs.