Long Noncoding RNA NEAT1Is Associated With The Spermatogenesis In Mice

Spermatogonial stem cells (SSCs) can self-renew to maintain their relative quantity, atthe same time can also differentiate to produce sperm which plays key roles in maintainingmale fertility. Spermatogenesis in a complicated process involving sequential expression ofmany genes. Long non-coding RNA (lncRNA) can regulate sequential gene expressions aswell as many biological functions. But lncRNA function in male reproduction is largelynuknown. lncRNA NEAT1is a core component of paraspeckle, which can mediateprocessing and translation of mRNA. NEAT1can influence fate decisions of humanembryonic stem cell through controlling sequcetial expression of key genes. The object ofthis study was to evaluate the funtion of NEAT1during spermatogenesis. The interferenceshRNA plasmid was successfully constructed. NEAT1-knockdown lentivirus was preparedand injected into the testicular seminiferous tubules of male mice. The main results were asfollows:(1) The bioinformatic analysis of NEAT1was conducted.(2) RT-PCR analysis showed that NEAT1was expressed in mouse testicular tissue andGC-1cells.(3) The shRNA sequence which can interference the expression of NEAT1was designedand its CD513B-U6interference shRNA plasmid was successfully constructed. Finally theinterference lentivirus which can knock down the expression level of NEAT1to69%wasacquired using the third generation of lentivirus packaging system.(4) The testis index from experimental group increased slightly compared to controlgroup but the change was not significant. The proportion of seminiferous tubules withspermatogenesis is86%, declined significantly compared with the control group (97%). Thisresults showed knocking down the expression level of NEAT1will affects spermatogenesis inmice.