The Study Of Molecular Regulatory Network And Related Candidate Genes In The Formation Of Chicken Cerebral Hernia

The brain,function as the nerve center in the animal body,controlling the physiological activities of organism directly or indirectly.Telencephalon is the most complex part of the brain,once a disease occurs,it will affect the normal life activities of the body.Cerebral hernia defined as a neurological disease in the chicken brain,which usually causing an abnormal behavior and low egg-laying rate in hens.However,little research focused on the formation of cerebral hernia in poultry.In this study,we aimed to compare the formation of telencephalon between chicken(phenotype of cerebral hernia)and chicken(phenotype of control);investigate the molecular network regulating telencephalon formation in transcriptome level,and provide a model to the study of brain disease both in chicken and human.This work will also provide a theoretical basis for further screening related candidate genetic markers to the poultry production.In this study,sample of brain from 28-days chicken(phenotype of cerebral hernia,n=3)and chicken(phenotype of control,n=3)was selected,and the molecular network of phenotypic difference between chicken(phenotype of control)and chicken(phenotype of cerebral hernia)was analyzed and revealed by transcriptome sequencing technique.Candidate genes and signaling pathways regulating the formation of chicken cerebral hernia are screened and identified primarily.This study explored the molecular regulation mechanism of cerebral hernia formation preliminarily and provided a theoretical basis for the breeding improvement of chicken in production.The main findings are as follows:1.The morphology(28-days)difference between cerebral hernia and control phenotype:In this study,the morphological differences of telencephalon in a specific period were compared by Nissl staining.It found that the cell body size of chicken(phenotype of control)telencephalon cells was significantly higher than that in mutant telencephalon tissue in hyperpallium.The above results were also confirmed by immunohistochemistry of Neu N,and it was found that the density and size of neuronal bodies in the hyperpallium of chicken(phenotype of cerebral hernia)telencephalon were significantly lower than those in chicken(phenotype of control).2.Molecular mechanism underlying of cerebral hernia formation in chicken(phenotype of cerebral hernia):(1)The transcriptional level differences of different phenotypes in telencephalon were compared and analyzed by transcriptome sequencing technique.A total of 337 differentially expressed genes were screened by bioinformatics analysis,of which 223 genes were up-regulated in chicken(phenotype of cerebral hernia)telencephalon,114 genes were down-regulated.(2)The enrichment analysis of differential genes by GO and KEGG,it showed that the results were mainly related to phosphorylation,vascular development,cell adhesion,cell proliferation,inflammatory response and other in GO analysis.Through KEGG enrichment analysis,we found that the differential genes were enriched in Wnt pathway,ECM-receptor interaction,Focal adhesion,m TOR pathway and other signaling pathways which related to cell development,cytoskeleton and neurological diseases.It speculated that THBS4,S100A10,GFAP,RICTOR and COL3A1 were candidate genes that regulate phenotypic differences in the telencephalon.Protein-protein interaction network showed that candidate genes were related to each other and regulate the abnormal development of telencephalon.(3)The expression pattern of candidate genes was detected by q PCR,and the results were consistent with transcriptome sequencing.The expression of THBS4,S100A10,GFAP,COL3A1 gene in chicken(phenotype of cerebral hernia)was significantly higher than that in chicken(phenotype of control),and the expression of RICTOR gene in chicken(phenotype of control)was significantly higher than that in chicken(phenotype of cerebral hernia).(4)In this study,it was found that there were significant morphological differences between wild type and chicken(phenotype of cerebral hernia)telencephalon in selected stage(28-days).Through transcriptional sequencing and bioinformatics analysis,it was found that the differentially expressed genes were mainly enriched and related to phosphorylation(CAV1?HSPB1?MMP2?PSEN1?TIMP3),vascular development(RNASE6?MMP2?ANXA2?CYR61?PSEN1?COL1A2?ANXA1),cell proliferation(ANXA1?ANXA2?CAV1?CEBPB?CYR61?EXFABP?HBEGF?HPGDS?MMP2?NFIB?S100A11),gliogenesis(NFIB?PSEN1?ANXA1)and inflammatory response pathways(ANXA1?CCL4?EXFABP?FN1?PSEN1).Through construction protein interaction network and combined with reports,candidate genes which highly associated with cerebral hernia were screened.The purpose of this study was to study the molecular network regulating chicken cerebral hernia in the preliminary transcription level,and to verify the key genes and signaling pathways controlling the formation of cerebral hernia.It provides a certain theoretical basis and evidence for the improvement of chicken breeds in the future.