Abstract

Cattleyak, a hybrid between cattle and yak, can adapt to the harsh environment of the highlands and also exhibit high production performance. However, its male infertility problem has not yet been solved, which limits the application of its hybrid vigor. In this work, we cloned the OTUD6A gene of cattleyak and analyzed its expression patterns and proliferative effect on Sertoli cells and Leydig cells. The expression level of OTUD6A in testicular tissues, Sertoli cells and Leydig cells were significantly lower than those in yak. Over-expression of OTUD6A in Sertoli cells and Leydig cells of cattleyak significantly increased their proliferative activity and upregulated the proliferation-associated genes. Furthermore, OTUD6A overexpression resulted in significant upregulation of tight junction-related genes ( TJP1 , TJP2 , CLDN11 , CLDN6 , OCNLD ) in Sertoli cells of cattleyak, and the expression of testosterone synthesis-related genes ( CYP11A1 and SF1 ) in Leydig cells were significantly downregulated, indicating that OTUD6A can influence cell function by upregulating the genes related to tight junctions in Sertoli cells and downregulating the genes related to testosterone synthesis in Leydig cells. These results indicate that the abnormal expression of OTUD6A in cattleyak may affect the functions of Sertoli cells and Leydig cells, thereby affecting spermatogenesis in cattleyak. This study provided a new theoretical basis for investigating the molecular mechanism by which OTUD6A regulates spermatogenesis in cattleyak and the molecular mechanism of spermatogenic arrest in cattleyak.