Es-ATG7 is required for spermatogenesis of Eriocheir sinensis and modulates p53-dependent apoptosis in germ cells

Abstract

Spermatogenesis is a complicated and highly regulated male gamete differentiation process that begins with the proliferation and differentiation of spermatogonia to the release of the mature spermatozoa. The autophagy-related gene <i>atg7</i> has been reported as closely related to spermatogenesis and communication of Sertoli cell-germ cells in mice, including acrosome biogenesis, sperm flagellum development, and ectoplasmic specialization assembly. However, the function of es-ATG7 and its molecular regulatory mechanism during spermatogenesis in Crustacea remain largely unknown. Here, we cloned and identified <i>es-atg7</i> from the testes of the Chinese mitten crabs <i>Eriocheir sinensis</i> and found that the expression of <i>es-atg7</i> was relatively high in testes through semi-quantitative RT-PCR. The dynamic localization of es-ATG7 detected by immunofluorescence may convey information about its role in the spermatogenesis of <i>E. sinensis</i>. Furthermore, a knockdown of <i>es-atg7</i> revealed that the malformed sperm with irregular sperm shape or loose nuclear cup and germ cell apoptosis were increased significantly. Accompanying this, we found an up-regulated expression of es-p53 during spermatogenesis in <i>es-atg7</i> knockdown groups. Altogether, our results indicate that es-ATG7 plays an essential role during spermatogenesis of <i>E. sinensis</i>, and we demonstrated that es-ATG7 acts as an antagonist for p53-dependent apoptosis induction in this process.