Pramef12基因在出生后小鼠睾丸精子发生过程中的功能研究

项目名称： Pramef12基因在出生后小鼠睾丸精子发生过程中的功能研究

项目编号： No.81501316

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 谢文海

作者单位： 山东理工大学

项目金额： 18万元

中文摘要： 由于男性不育导致的不孕不育问题逐年递增，男性生殖健康受到越来越多的关注。人类精子发生过程涉及4000多个基因的表达，任何一个相关基因的异常表达都会引起精子发生障碍。通过前期高通量实验，我们发现了一个与精原细胞凋亡相关的基因Pramef12，随着睾丸的发育动态表达，过表达Pramef12能够增强精原干细胞对凋亡诱导试剂的敏感性。在此基础上，本项目拟在体外通过在精原干细胞中过表达和沉默Pramef12研究其功能；比较过表达、沉默与对照组精原干细胞的表达谱数据寻找差异基因并预测Pramef12调控网络；在体内通过构建Pramef12条件敲除小鼠验证其功能。采用生物信息学预测与传统分子生物学实验相结合的方法，探究Pramef12参与调控精原细胞凋亡的分子机制。本研究将有助于我们了解精子发生过程中精原细胞凋亡的分子机制，为男性不育疾病的诊断、治疗和预防提供新的认识和理论依据。

中文关键词： Pramef12基因；精子发生；细胞凋亡；转录调控；基因敲除

英文摘要： Male reproductive health has attracted increasing attention due to the annual growth of male infertility. More than 4000 genes are involved in the process of human spermatogenesis. Any abnormal expression of the related genes will cause spermatogenesiss disturbance. According to the previous high throughput experiment, we found that pramef12 is related to the apoptosis of spermatogonia. The expression of pramef12 changes dynamically during the development of testis. Overexpression of pramef12 enhances the sensitivity of spermatogonial stem cell to apoptosis-inducing agent. Based on those findings, we plan to study the function of pramef12 in vitro by overexpression and knock-down in spermatogonial stem cell. Differently expressed genes are found by comparing gene expression profiles from overexpression group and knock-down group to control group respectively. The regulation network of pramef12 will be predicted by the interactions among those different expressed genes. The function of pramef12 is verified by constructing pramef12 germline conditional knockout mice in vivo. The molecular mechanisms of pramef12 involved in apoptosis of spermatogonia will be further elucidated by combining bioinformatics prediction methods and traditional molecular biological experiments. This study will help us to understand the molecular mechanisms of spermatogonia apoptosis during spermatogenesis and provide new insights and theoretic foundation for clinical diagnosis, treatment and prevention of male infertility.

英文关键词： Pramef12;Spermatogenesis;Cell apoptosis;Transcriptional regulation;Gene knockout