Effects of SPATA6 on proliferation, apoptosis and steroidogenesis of Hu sheep Leydig cells in vitro.

This study aimed to investigate the expression pattern of spermatogenesis associated protein 6 (SPATA6) in Hu sheep testis and to ascertain the effects of SPATA6 on sheep Leydig cells (LCs) function linked to spermatogenesis. In the present study, we detected a 1970 bp cDNA fragment of SPATA6 included a 1467 bp coding sequence which encoded 487 amino acids. Meanwhile, sheep SPATA6 shared 51.70%-97.41% amino acid sequences with its orthologs compared with other species. In addition, SPATA6 was highly expressed in testis and localized in cytoplasm and nucleus of LCs as well as spermatogenic cells at different stages. Compared to the negative control (NC), SPATA6 interference promoted apoptosis of LCs with the increase of BAX/BCL-2 mRNA and protein levels, while the results of SPATA6 overexpression were on the contrary. Meanwhile, cell cycle was blocked at G2/M phase and CDK1 and CCNB1 were down-regulated after SPATA6 interference. SPATA6 overexpression induced cell cycle transfer G0/G1 into S and G2/M phase with upregulation of CDK1, CDK4, CCND1 and CCND2. Moreover, the secretion of testosterone hormone and the expression of StAR in LCs with SPATA6 overexpression were significantly promoted. Overall, our data suggest that SPATA6 is an important functional molecule of spermatogenesis, via regulating the proliferation, apoptosis and testosterone biosynthesis of Hu sheep LCs. These findings will enhance the understanding of the roles of SPATA6 in sheep spermatogenesis.