Investigating a novel role of ejaculate RNA in fertility

项目摘要 内分泌生物的雄性射精是多种成分的混合物，包括精子和 在交配过程中转移到雌性体内的许多其他大分子。关于男性贡献的研究 对生育力的研究主要集中在精子上，最近则集中在精液中的蛋白质上。虽然这些 研究极大地推进了我们对交配后相互作用的理解， 特发性不孕症表明，对精液中未充分研究的分子的研究可能会产生新的见解， 转化为新的调节生殖结果的因素。我们之前发现雄性的RNA转移 在亚利桑那果蝇交配时对雌性产生了影响。在这份提案中，我们提供了通过资助收集的证据 我们最近获得的R21奖项证明了雄性RNA转录本在D.亚利桑那 女性生殖道在这项工作的基础上，拟议研究的目标是建立功能 精液RNA对雌性交配后反应的影响（目的1），研究它们在交配后反应中的作用。 女性（目标2），并确定来源和机制，它们被包装在男性（目标3）。我们 利用我们的D的独特方面。arizonae/亚氏拟谷盗D.莫哈文斯研究系统，联合收割机，使一个基因 一个易于处理的，操纵性的实验系统，特别适合于研究男性的功能， 女性生殖道内的RNA。有了这个强大的系统，我们联合收割机结合蛋白质组学分析，使用我们的 最近开发的变体辅助SILAC蛋白质组学分析（VASPA）方法， mdFTPs（男性衍生的女性翻译蛋白）。鉴于RNA似乎是一种保守的 男性射精的特征，我们希望我们的发现将有广泛的意义，更好地了解 包括人类在内的一系列物种交配后的相互作用。

PROJECT SUMMARY The male ejaculate of internally fertilizing organisms is a diverse mixture of components including sperm and numerous other macromolecules that are transferred to females during mating. Research on male contributions to fertility has mainly focused on sperm, and, more recently on proteins present in the seminal fluid. While these studies have greatly advanced our understanding of postcopulatory interactions, the relatively high incidence of idiopathic infertility suggests that investigation of understudied molecules in the ejaculate may yield novel insights into novel factors mediating reproductive outcomes. We previously discovered the transfer of RNA from males to females during copulation in Drosophila arizonae. In this proposal, we provide evidence gathered with funding from our recent R21 award demonstrating translation of male RNA transcripts within cells of the D. arizonae female reproductive tract. Building off this work, the goals of the proposed research are to establish functional effects of seminal fluid RNAs on the female postmating response (Aim 1), investigate how they function in females (Aim 2), and identify the source and mechanisms by which they are packaged in males (Aim 3). We capitalize on unique aspects of our D. arizonae/D. mojavensis study system that combine to make a genetically tractable, manipulative experimental system that is particularly well suited to investigate the function of male RNA within the female reproductive tract. With this powerful system we combine proteomic analyses using our recently developed Variant Assisted SILAC Proteomic Analysis (VASPA) approach with detailed investigations of individual mdFTPs (male-derived female translated proteins). Given that RNA appears to be a conserved feature of male ejaculates, we expect our findings will have broad significance for better understanding postcopulatory interactions across a range of species, including humans.