褪黑素及其受体调控绵羊精原干细胞分化为单倍体精子细胞机制的研究

项目名称： 褪黑素及其受体调控绵羊精原干细胞分化为单倍体精子细胞机制的研究

项目编号： No.31501953

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 畜牧学与草地科学

项目作者： 邓守龙

作者单位： 中国科学院动物研究所

项目金额： 20万元

中文摘要： 本研究组建立了绵羊SSCs/睾丸体细胞离体分化系统，可分化出功能性单倍体精子(显微注射成熟卵母细胞后，可获得囊胚)。但绵羊SSCs分化为单倍体精子的效率较低。研究表明短日照动物绵羊睾丸中存在褪黑素(MLT)受体。本研究通过添加外源MLT可提高SSCs体外分化为精子细胞的能力，但其作用机制不明确。阐明精子发生机制是调控SSCs体外分化为单倍体精子的关键。本研究将重点探索MLT及其受体调控绵羊精子发生的分子机制。确定MLT及其受体在睾丸和SSCs分化系统中的来源与定位。通过在建立的分化系统中添加MLT及其受体抑制剂，检测睾丸体细胞内睾酮与cAMP含量、相关激素受体表达与抗氧化凋亡相关因子变化；并通过活体埋植MLT，进行睾丸转录组测序，分析体内精子发生相关基因的变化，明晰MLT通过什么信息通路调控精子发生。从而为阐明绵羊精子发生机制和应用SSCs体外分化出单倍体精细胞进行转基因研究提供理论依据。

中文关键词： 精原干细胞分化；褪黑素；单倍体精子；精子发生；绵羊

英文摘要： We have established an in vitro sheep SSCs/testis somatic cells co-culture system to produce haploid sperms, which were microinjected into mature oocytes and hatched blastocysts. However, the efficiency of sheep haploid sperm production solution was much low. Studies have shown that melatonin receptor present in the testicular cell of ovine (a short day breeding species). The present study was designed to explore the molecular mechanism of MLT effect on sheep SSCs/testis somatic cell system differentiation efficiency in vitro in presence or absence of MLT or its receptor inhibitors. The key point is to confirm cell source of MLT/MLT receptor in the testis and in the co-culture system, and to clarify the involvement of signaling pathways. These studies will provide academic basis for the mechanism of ovine spermatogenesis, and the transgenic studies using the germ cells produced by the in vitro SSCs/testis somatic cell differentiation system.

英文关键词： SSCs differentiation;Melatonin;Haploid spermatids;Spermatogenesis;Sheep