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.

项目总结 体内受精有机体的雄性射精是一种多种成分的混合物，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 对生育能力的研究主要集中在精子上，最近又集中在精液中的蛋白质上。而这些 研究极大地提高了我们对交配后相互作用的理解，相对较高的发病率 特发性不孕症提示，对射精中研究不足的分子的研究可能会产生新的见解。 转化为调节生殖结果的新因素。我们之前发现了从男性身上转移RNA 在果蝇交配过程中传给雌性。在这项提案中，我们提供了用资金收集的证据 从我们最近的R21奖中证明了雄性RNA转录本在Arizonae细胞内的翻译 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 精液RNA对雌性交配后反应的影响(目标1)，研究它们在 雌性(目标2)，并确定它们在雄性体内的来源和机制(目标3)。我们 利用我们的D.arizonae/D.mojvensis研究系统的独特方面，这些研究系统结合在一起，在遗传上 易于操控的实验系统，特别适合研究男性的功能 雌性生殖道内的核糖核酸。有了这个强大的系统，我们使用我们的 新近发展的变异辅助SILAC蛋白质组学分析(VASPA)方法及其详细研究 个体mdFTPs(男性来源的女性翻译蛋白)。鉴于RNA似乎是一种保守的 关于男性射精的特征，我们希望我们的发现将对更好地理解 交配后的相互作用跨越了一系列物种，包括人类。