长链非编码RNA NEAT1对小鼠精子发生的影响

Effect of Long Noncoding RNA NEAT1 on Spermatogenesis in Mice

为研究长链非编码RNA NEAT1对小鼠精子发生的影响,构建针对NEAT1的干扰载体并包装慢病毒,通过将干扰慢病毒注射到成年小鼠睾丸曲细精管和定量PCR,H.&E.染色等方法观测NEAT1对小鼠精子发生的影响。研究结果表明,NEAT1在睾丸组织和GC-1生殖细胞系中表达;成功构建CD513B-U6-NEAT1干扰载体,利用第三代慢病毒包装系统包装获得干扰慢病毒且干扰效果明显(NEAT1转录本水平下调69%);慢病毒注射试验结果显示干扰病毒注射组睾丸指数轻微增加,有精子发生的曲细精管比例由97%减少为86%(P<0.05),表明小鼠精子发生受到影响,NEAT1具有维持雄性小鼠精子发生的作用。

To study the the effect of long noncoding RNA NEAT1 on spermatogenesis in mice,an interference shRNA plasmid and lentivirus was successfully constructed.A method which combined quantitative PCR,H.E.staining and injection of lentivirus into the the seminiferous tubules was applied to detect its effect on the whole process of spermatogenesis in mice.The results showed that NEAT1 expressed in testicular tissue and GC-1spermatogonial cell lines;The design of interference NEAT1 shRNA and constrution of CD513B-U6 interference plasmid were successful.Interference lentivirus was acquired using the third generation of packing system and the interference effect was obvious.The testicular index of interference lentivirus injection group increased slightly when campared to the control group and the percentage of seminiferous tubule with spermatogenesis declined（P〈0.05）,indicating the spermatogenesis process in mice was affected by NEAT1.In conclusion,NEAT1 regulated spermatogenesis in male mice.