Function Study Of Extracellular Vesicles And LncRNA5512 On Spermatogenesis In Mice

Objective: The spermatogenesis of mammalian male individuals is a process that is precisely regulated in many ways.A large number of regulatory mechanisms during spermatogenesis are not yet clear.Exploring the regulation mechanism during spermatogenesis is of great significance for the diagnosis and treatment of male infertility.We found that there are extracellular vesicles associated with spermatogonial stem cells near the basement membrane of mammalian testis.We explored the regulatory mechanisms during spermatogenesis.In addition,by sequencing mouse transcriptomes,we discovered a new long non-coding RNA which is specifically and highly expressed in the testis,we named it lncRNA5512 and study its function in spermatogenesis in mice.Methods and Results: 1.Primary Sertoli cells were isolated by two-step enzyme digestion.The extracellular vesicles of the testis were obtained by ultracentrifugation,and identified using immunoelectron microscopy,Nanosight,Western Blot,etc.2.Western Blot and q RT-PCR were used to detect the change of GDNF of primary Sertoli cells after testicular extracellular vesicle treatment.It was found that Thy1 + testicular extracellular vesicles can downregulate Sertoli cell GDNF expression.3.CRISPR / Cas9 was used to construct lncRNA5512 knockout mice,and the changes in fertility-related indicators were detected.It was found that lncRNA5512 deletion had no significant effect on spermatogenesis and fertility in male mice.Conclusions: 1.Thy1 + extracellular vesicles from testis inhibit the expression of GDNF in Sertoli cells in vitro.2.The deletion of lncRNA5512 specifically expressed in spermatocytes and round spermatids has no significant effect on spermatogenesis and fertility of male mice.