睾丸富集表达的X染色体连锁的长链非编码RNA的鉴定和功能研究

Spermatogenesis in testis is a very complicated but coordinated process, which includes mitosis, meiosis and spermiogenesis. Eventually diploid spermatogonia are differentiated into haploid round spermatids. This process is not only controled by hormones and transcription factors , but also by noncoding RNAs including miRNAs, piRNAs and long noncoding RNAs ( LncRNAs). So far the roles of LncRNAs in spermatogenesis are largely unknown. In our preliminary study, we performed large scale RNA-sequencing analysis on various adult mouse tissues including serveral male reproductive organs such as testis, epididymis, prostate and seminal vesicle as well as 6-day-old mouse testis. We have identified two X-chromosome linked LncRNA clusters, which contained those lncRNAs with enriched expression in testis and without polyA tail. In this study, We will perform a series of in-vitro and in-vivo studies on these two clusters. We will identify their expression pattern in various tissues and various developmental stages as well as various spermatogenic cells and confirm if these lncRNAs escape meiotic sex chromosome inactivation and identify their function. This original study will help to elucidate the role of these novel lncRNAs in spermatogenesis and provide clues for diagnosis and therapy of male infertility.

精原干细胞经过有丝分裂，减数分裂和精子变形三个阶段，产生有尾巴的单倍体精子。这一系列复杂过程既受到激素和转录因子调控，也受到非编码RNA包括长链非编码RNA(LncRNA)调控。目前LncRNA在精子发生中的作用基本上未知。本项目组前期利用去核糖体RNA二代测序技术，对成年小鼠不同组织包括睾丸，前列腺，附睾，精囊腺和新生第6天睾丸进行RNA测序，从中鉴定到一大批睾丸富集表达lncRNA，包括两个X染色体连锁的睾丸富集表达的LncRNA簇，其中大都不带polyA尾。在本项目研究中，我们将深入研究这两个lncRNA簇，包括组织分布和不同发育时期及各级生精细胞表达研究，确定它们是否逃脱减数分裂性染色体失活，从体外和体内两方面研究它们功能。 通过本项目研究，将有助于揭示这些新类型LncRNA在精子发生中作用，为男性不育的诊治提供新思路。

精子发生从精原干细胞开始经历有丝分裂，减数分裂和精子变形，产生有尾巴的单倍体精子。这一复杂过程既受到激素和转录因子调控，也受到非编码RNA包括长链非编码RNA(LncRNA)调控。 目前 LncRNA在精子发生中的作用基本上未知。本项目组利用去核糖体RNA二代测序技术，对成年小鼠不同组织进行RNA测序，从中鉴定到一大批睾丸富集表达lncRNA，包括两个X染色体连锁的睾丸富集表达的LncRNA簇，其中大都不带polyA尾，有可能逃脱了减数分裂时性染色体失活。我们深入研究这些lncRNA的表达特性并用基因敲除的方法研究功能。另外，我们还对一类特殊的LncRNA-环状RNA在睾丸中的表达进行了分析，鉴定到大量睾丸特异表达的环状RNA。本研究丰富了我们对睾丸中的lncRNA和环状RNA的理解，为进一步深入研究精子发生过程的调控机制奠定基础。在本研究支持下，共发表研究论文和综述8篇(其中SCI论文2篇), 培养硕士研究生毕业两名。