NYD-SP27调控绵羊精子发生及获能的分子机制

项目名称： NYD-SP27调控绵羊精子发生及获能的分子机制

项目编号： No.31460683

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 赛务加甫

作者单位： 石河子大学

项目金额： 52万元

中文摘要： PLCζ在生殖发育中发挥重要，其对精子的发生发育及卵母细胞激活等相关研究日益成为生殖发育的研究热点。然而，PLCζ家族中的睾丸发育特异性蛋白-27基因( NYD-SP27)，却被认为是一种新的精子内源性去能因子。已证实NYD-SP27在小鼠精子获能上具有重要作用，但NYD-SP27在绵羊精子获能中的作用未见相关报道。本项目的研究思路为：筛选绵羊NYD-SP27基因的有效shRNA构建干扰表达载体，用脂质体包被后注入绵羊睾丸组织中，检测NYD-SP27表达下调对绵羊精子发生及获能的影响；构建绵羊NYD-SP27基因的真核表达载体用脂质体包被后注入绵羊睾丸组织中，检测NYD-SP27的过表达对精子发生发育及精子相关信号调控通路的影响，并研究NYD-SP27在睾丸或精子中的表达和定位，为揭示NYD-SP27在精子发生及获能中的作用机制提供一定的科学依据。

中文关键词： 绵羊；睾丸发育特异性蛋白-27；磷脂酶C；zeta；精子获能；精子发生

英文摘要： PLCζ plays an important role in reproductive development. The effect on early spermic occuring and development and the mechanism of phospholipase C zeta (PLCζ) for egg activation is becoming a hot topic in studies of reproductive development. However, the specific protein gene 27(NYD-SP27) in the development of testis has been identified as a new intrinsic decapacitation factor (DF) in sperm, which belongs to the PLCζ family. It has been confirmed that NYD-SP27 plays a pivotal role in the regulation of sperm capacitation in mouse, but there are no related reports on sheep so far.The research idea of this study is to screen effective shRNA of sheep NYD-SP27 gene to construct interference expression vectors,and coated it with liposomes ,and then the recombinant plasmid was injected into the sheep testicular tissue ,to detect the effects of down-regulated of NYD-SP27 on sheep spermatogenesis and sperm capacitation;, to detect Eukaryotic expression vector for NYD-SP27 gene was constructed, recombinant plasmid which peridiumed by lipofectamine was injected into the sheep testicular tissue to detect whether over-expressing of NYD-SP27 inhibit the expression of PLCζ in testicular tissue or sperm and its localization. The study would provide a scientific basis for revealing the functionatin of NYD-SP27 for spermatogenesis and sperm capacitation。 To detect the influence on spermatogenesis and regulatory pathway of related signals by the over-expressing of NYD-SP27,and to study the NYD-SP27 expression and localization in the testes or the sperm.

英文关键词： Sheep;NYD-SP27;phospholipaseC zeta;spermcapacitation;spermatogenesis