Investigating a novel role of ejaculate RNA in fertility

研究射精 RNA 在生育中的新作用

• 批准号: 10718225
• 负责人: Jeremy Matthew Bono
• 金额: $ 60.92万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-18 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10718225
• 关键词: Area; Automobile Driving; Behavior; Biological Assay; Cells; Complex; Copulation; Drosophila genus; Ejaculation; Exploratory/Developmental Grant for Diagnostic Cancer Imaging; Female; Fertility; Funding; Goals; Human; Immunity; Incidence; Individual; Insect Control; Insect Vectors; Insecta; Investigation; Life; Mammals; Mediating; Molecular; Organism; Partner in relationship; Play; Proteins; Proteomics; RNA; Reporting; Reproduction; Reproductive Physiology; Research; Role; Seminal fluid; Source; System; Tissues; Transcript; Transfer RNA; Translating; Translations; Trees; Variant; Work; clinically significant; design; exosome; extracellular vesicles; human disease; human pathogen; idiopathic infertility; insight; interest; macromolecule; male; novel; novel therapeutics; offspring; protein function; reproductive; reproductive outcome; reproductive tract; response; sex; sperm cell

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.

项目概要 体内受精生物体的雄性精液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。男性贡献研究 对生育力的影响主要集中在精子上，最近又集中在精液中的蛋白质上。虽然这些 研究极大地增进了我们对性交后相互作用的理解，即相对较高的发生率 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 研究介导生殖结果的新因素。我们之前发现了雄性RNA的转移 亚利桑那果蝇交配期间对雌性的影响。在本提案中，我们提供了通过资助收集的证据 来自我们最近的 R21 奖，展示了亚利桑那州 D. arizonae 细胞内雄性 RNA 转录本的翻译 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性的 精液 RNA 对雌性交配后反应的影响（目标 1），研究它们如何在 女性（目标 2），并确定它们在男性中的来源和包装机制（目标 3）。我们 利用我们的 D. arizonae/D. 的独特方面。莫哈文西斯研究系统结合起来形成一个基因 易于处理的、可操作的实验系统，特别适合研究男性的功能 女性生殖道内的 RNA。借助这个强大的系统，我们使用我们的技术将蛋白质组分析结合起来 最近开发的变异辅助 SILAC 蛋白质组分析 (VASPA) 方法进行了详细研究 单个 mdFTP（雄性衍生的雌性翻译蛋白）。鉴于 RNA 似乎是保守的 男性射精的特征，我们希望我们的发现对于更好地理解具有广泛的意义 包括人类在内的一系列物种的交配后相互作用。