Studies On The Genome-wide DNA Methylation Changes And Transcriptomic Profiling Of Testis In Weizhu Boars

Testicular development and spermatogenesis after the initiation of puberty in male mammals is an important physiological process and is of great significance for the study of reproductive health.DNA methylation is one of the most common epigenetic modifications in organisms,regulating the expression of genes and thus affecting animal physiology.In this study,the testis of the Weizhu boars before and after the puberty(75 days old and 110 days old)was used as the material.The histological morphology of the testis was observed by tissue section,and whole-genome bisulfite sequencing(WGBS)and RNA-seq technology combined with bioinmatic tools were used to study methylation and transcriptome changes,screen differential methylation and differentially expressed genes.Through the combined analysis of methylation and transcriptome data,we select some candidate genes and key pathways.It provides a theoretical basis for revealing the genetic mechanism of testicular development and spermatogenesis in warts.The research results are as follows:(1)From the tissue section of the testis of the porcine pig,it can be seen that the 75-day-old boar has not yet produced sperm,and the testicular tissue structure of the 110-day-old boar has basically developed,similar to the mature boar,combined with the performance of the boar.75 days and 110 days of age can be used as two time points before and after the puberty.(2)There are 13373 regions in the differentially Methylated Regions(DMR)in two periods.The anchoring genes are carried out in the DMRs region.A total of 3,330 genes are found,and the methylation levels of different functional regions of the genome,promoter and internal The inclusions and 3’UTR(Untranslated Regions)were higher than the 5’UTR and exon regions,and a similar situation was found for methylation density analysis.In this study,we used GO enrichment analysis to find a number of cellular component enrichment related to the biological functions of the study,including protein binding,anatomical structure development,regulation of GTPase activity,blood vessel development and so on,which are involved in testicular development.The axon guidance pathway was only found in KEGG as a significant associated.(3)More than 12,700 transcripts were detected by RNA-seq.Through detection and screening(|log2.foldchange|≥1,p<0.05),239 up-regulated genes and 300 down-regulated genes were obtained.we used GO enrichment analysis to find a number of cellular component enrichment related to the biological functions related to testicular development and spermatogenesis,including biological regulation,regulation of cellular processes,cell differentiation,reproductive process,gamete generation,spermatogenesis,male gamete generation,and so on.Pathway enrichment analysis was led to protein digestion and absorption,metabolic pathways,GnRH signaling pathway,and estrogen signaling pathway,ECM-receptor interaction and so on.(4)Through differential genome-binding analysis of genome-wide methylation and transcriptomics,a total of 197 differential genes were found for GO enrichment,and the GO pathway of testicular development and spermatogenesis-related biological processes was found to have ATP hydrolytically coupled protons.ATP hydrolysis coupled proton transport,ATP hydrolysis coupled transmembrane transport,regulation of Wnt signaling pathway,and negative regulation of Wnt signaling pathway,regulation of epithelial cell migration,Wnt signaling pathway,cell-cell signaling by wnt,and the like.Pathway enrichment analysis of differential genes yielded 35 pathway enrichment pathways.These pathways are associated with testicular development and spermatogenesis,including the Wnt signaling pathway,the cAMP signaling pathway,the PI3K-Akt signaling pathway,ECM-receptor interaction,insulin secretion,focal adhesion and so on.At the same time,28 genes with differentially methylated and differentially expressed promoter regions were found.