The Effect of Splicing Factor hnRNPF on Spermatogenesis in Mongolian Horse
The aim of this study was to investigate the effect of alternative splicing factor heterogeneous nuclear ribonucleoprotein F(hnRNPF)on spermatogenesis in Mongolian horses.Testicular sertoli cells were isolated and cultured from 3-year-old Mongolian horse.The hnRNPF overexpression lentiviral vector was constructed and transfected into the sertoli cells of Mongolian horse testis.CCK-8 was used to detect the proliferation of cells at 0 h,24 h,48 h and 72 h after transfection,and the optimal infection time was determined with 3 duplication per group.The RNA of transfected and untransfected Mongolian horse sertoli cells was extracted,and the primers of genes known to be related to spermatogenesis were designed for qRT-PCR to detect the expression of spermatogenesis-related genes in the two types of cells.The results of agarose gel electrophoresis showed that the hnRNPF lentivirus overexpression vector was successfully constructed.CCK-8 assay showed that the cells transfected with the expression lentiviral vector had the highest activity at 72 hours,and the activity of transfected cells was higher than that of untransfected cells,indicating that the overexpression of hnRNPF could promote the proliferation of Mongolian horse sertoli cells.qRT-PCR results showed that the expression levels of spermatogenesis-related genes(PKMYT1,CDC25C,YWHAZ,BUB1,BTRC,CCNE1,CALM1,PLK1,REC8,MAPK3,and ADCY7)were significantly different between the two types of cells.All were highly expressed in cells transfected with hnRNPF overexpression vector.The results suggest that hnRNPF may directly or indirectly promote spermatogenesis in Mongolian horses,which provides a new idea for improving the sperm count and semen quality of Mongolian horses.
万方数据
维普
