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.

项目摘要 内部受精生物的雄性射精是包括精子和包括精子和 在交配过程中转移到女性的许多其他大分子。男性贡献的研究 生育能力主要集中在精子上，最近，最近出现在精确液中的蛋白质上。而这些 研究极大地提高了我们对填充后相互作用的理解，这是相对较高的发生率 特发性不育症表明，对射精中研究的分子的研究可能会产生新的见解 成为介导生殖结果的新因素。我们以前发现RNA从男性转移 在果蝇亚利桑那州的交配过程中向女性致敬。在此提案中，我们提供资金收集的证据 从我们最近的R21奖中展示了D. arizonae细胞内男性RNA转录本的翻译 女性生殖道。在这项工作中，拟议的研究的目标是建立功能 开创性液体RNA对雌性后反应反应的影响（AIM 1），研究它们如何在 女性（AIM 2），并确定其包装在男性中的来源和机制（AIM 3）。我们 利用D. Arizonae/d的独特方面。 Mojavensis研究系统结合起来制作遗传 可进行操作的操纵实验系统，特别适合研究男性的功能 女性生殖道中的RNA。使用这种强大的系统，我们使用我们的蛋白质组学分析结合了 最近开发的变体辅助SILAC蛋白质组学分析（VASPA）方法以及详细研究 单个MDFTP（男性衍生的女性翻译蛋白）。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将具有广泛的意义，以更好地理解 包括人类在内的各种物种之间的填充后相互作用。