Expression Of 12 MicroRNAs And Target Genes In Xiang Pig Testis Tissue

The present paper chosen twelve microRNA as candidates based on the sequencing data of microRNAs software in testes of Xiang pig.The target genes of twelve miRNAs were predicted by the miRSystem and used to analysis the GO and KEGG through Kobas and DAVID databases.The changes of twelve miRNAs level were determined by RT-qPCR in different stages of porcine testes.At the same time,seven target genes associated with reproduction trait were verified.The main results of this study were as follows:(1)Target genes of 543 microRNAs were deduced.A total of 2834 targets of twelve microRNA were checked out using miRsystem software.These genes were gathered into 3685 GO entries and 34 signaling pathways by GO and KEGG analysis,which involved in the biological processes including testicular development and spermatogenesis.It was interested that several targets of twelve miRNAs were enriched in the MAPK signaling pathway.(2)According to the histological changes of the testes,six groups of Xiang pig were divided into four stages including immature in one month old,puberty in 2 ~ 3month old,sexual maturity in 4 ~ 6 month old and old age in 48 month.(3)By using a stem-loop primer for reverse transcription,the expression levels of twelve microRNAs in testis were much diversity.A total of ten miRNA showed negative control for the immature testes,which were miR-222-3p,miR-16-5p,let-7f-5p,miR-181b-5p,miR-424-5p,miR-23a-3p,miR-185-5p,miR-29a-3p,miR-29c-3p,miR-1-3p.Three ones,let-7f-5p,miR-222-3p and miR-29c-3p,effected in puberty testes of Xiang pig.In stage of sexual maturation,the important molecular were mi R-449b-5p,miR-16-5p,let-7f-5p,miR-140-3p,miR-29c-3p and mi R-1-3p,while it was miR-185-5p,miR-222-3p,mi R-29c-3p and miR-1-3p in old age of testes.(4)When both of miR-16-5p and miR-449b-5p reached the highest expression level,it could be taken as a molecular marker for sexual maturity of the testis.The highest value miR-222-3p suggested that the testis reached the old stage for elimination.(5)The expression levels of MAPK1 and MAP2K1 genes in testes of 2 and 6months were much higher than the others.(6)It was miR-449b-5p that might effect on the spermatogenesis from immature to mature stages of testes by the MAPK signal pathway.In this study,we analyzed the developmental changes of twelve miRNAs in porcine testis.The results would be helpful for the understanding of the miRNA regulation mechanism of testis development and for the breeding of male pig.