| The process of testicular development and spermatogenesis needs the synergistic roles of a variety of cell types including germ cells,Leydig cells and Sertoli cells,which is controlled by extremely complex and strict molecular spatiotemporal regulation.However,the understanding of molecular mechanisms underlying testicular development remains largely unclear.Recently,as a new kind of long chain noncoding RNA,circular RNA has been found in testis or seminal fluid of human and cattle and may be involved in the regulation of testicular development and spermatogenesis.However,the expression profile and potential biological function of circular RNA in the testis of boar are not clear.N6-methyladenosine(m6A)modification is the most common modification in eukaryotic mRNA,which is involved in many physiological and pathological processes,such as cell differentiation,growth,development,and ontogeny.However,little is known about the relationship between m6A and testicular development and spermatogenesis.In this study,we selected testicular tissues of Landrace boar at different developmental stages and used high-throughput sequencing technology and bioinformatics to reveal the expression characteristics and expression differences of circular RNA and m6A in testis at different stages.Objective to predict and analyze the potential biological functions of differentially expressed circular RNA and m6A modification in testicular development and spermatogenesis.The discovery of its potential regulatory genes and signaling pathways may provide us with new clues and theoretical hypotheses for the study of spermatogenesis,testicular development,and male animal reproduction.H&E staining showed that the seminiferous tubules of testis of D30 boars were not fully developed,and there was no obvious lumen in testis;The testicular volume of D210 boars increased significantly,the seminiferous tubules developed completely,contained a large number of different types of germ cells,and produced a large number of sperm.Total RNA-seq was used to detect the expression profile of circular RNA in the testis before and after sexual maturation.High throughput sequencing revealed that there was abundant circular RNA in testis.34521 and 31803 circular RNA were identified in testis before and after sexual maturation.Bioinformatics analysis showed that these circular RNAs were widely distributed on autosomal and sex chromosomes,and their length was mainly between 100 and 10000 nt(58.97%),mainly from the exon region of the source genes.Most of the circular RNAs were formed by 1-7 exons(86.54%),and some of the source genes could produce multiple circular RNAs.2326 differentially expressed circular RNAs from 1526 source genes were found during testicular maturation in boars,of which 1003 were up regulated and 1323 down regulated in the testicular tissues of D210 boars.In addition,GO function analysis of source genes of differentially expressed circular RNA showed that these circular RNAs were mainly related to spermatogenesis,positive regulation of hormone biosynthesis process,germ cell development,male meiosis,and other biological processes.KEGG pathway enrichment analysis showed that they were involved in stem cell pluripotency regulation,tight junction,adhesion junction,camp and other signaling pathways.Therefore,the above results indicate that there are a large number of circular RNAs expressed in the boar testis before and after sexual maturity,which show a developmental stage specific dynamic expression pattern,and they are closely related to biological processes such as testicular development and spermatogenesis.These results will provide a potential molecular marker for testicular development and selection of elite boars.To study the dynamic changes of the methylation status during the development and maturation of the testis,the whole transcriptome m6A map of was measured before and after the sexual maturity.We describe the wide distribution pattern of m6A methylation modification in testis development and analyze the relationship between gene expression and m6A methylation modification,and identify the genes of differences m6A methylation modification,which may be related to the biological function differences of testis before and after sexual maturity.These results provide a basis for determining the presence and map of m6A methylation in testis and expand our understanding of the role of m6A in mammalian organ development and growth. |
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