Beyond spermatogenesis: Additional roles of seminal fluid fatty acids in male fertility and age-related reproductive success

超越精子发生：精液脂肪酸在男性生育力和与年龄相关的生殖成功中的其他作用

• 批准号: 10526745
• 负责人: Hock Wee Henry Chung
• 金额: $ 24.34万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-15 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10526745
• 关键词: Address; Affect; Age; Aging; Animal Model; Animals; Assisted Reproductive Technology; Biochemical Pathway; Candidate Disease Gene; Cell membrane; Chemicals; Development; Diagnosis; Drosophila genus; Drosophila melanogaster; Environment; Europe; Fatty Acids; Female; Fertility; Genes; Genetic; Goals; Health; Human; Incidence; Infertility; Invertebrates; Knock-out; Knowledge; Lead; Lipids; Male Genital Organs; Male Infertility; Mammals; Mass Spectrum Analysis; Mediating; Membrane; Methods; Modeling; Names; Organ; Outcome; Partner in relationship; Pathway interactions; Play; Production; Proteins; Publishing; RNA Interference; Reproduction; Reproductive Health; Reproductive Physiology; Research; Role; Seminal fluid; Spermatocytes; Spermatogenesis; System; Testing; Testis; Tissues; Validation; age related; aged; differential expression; enzyme biosynthesis; fatty acid biosynthesis; fatty acid elongases; fertility improvement; gene synthesis; genetic manipulation; idiopathic infertility; improved; innovation; insight; knock-down; lipid biosynthesis; lipidome; lipidomics; male; male fertility; novel; reproductive success; sperm cell; sperm viability; tool; transcriptome; transcriptomics

Project Summary/Abstract In many animals, the seminal fluid plays a major role in male reproductive success. Proteins in the seminal fluid have significant effects on multiple features of male and female reproductive physiology. However, relatively little is known about a second major component of seminal fluid, lipids, beyond their roles as sperm membrane constituents. The overall objectives in this proposal are to i) identify differences in the lipidomic and transcriptomic profiles of seminal fluid in young and aged males; and ii) directly test how manipulating the lipid profile of seminal fluid impacts male reproductive success. This proposal will address the hypothesis that the seminal fluid lipidome influences male fertility and changes with age, thus contributing to the decline of aging-related reproductive success. This hypothesis is supported by the applicants’ published studies showing that fatty acids and related lipids in the seminal fluid of Drosophila males are essential for male fertility. Both transposon-mediated knockout and RNAi knockdown of a key fatty acid biosynthesis enzyme in the male ejaculatory bulb (EB), an organ that is not involved in spermatogenesis, causes a significant decrease in male fertility, strongly supporting the contribution of seminal fluid fatty acids to male reproductive success. In contrast to mammalian models, use of Drosophila allows seminal fluid lipids produced in the EB to be manipulated without interfering with spermatogenesis in the testes. Powerful genetic manipulation in combination with lipidomic and transcriptomic analysis will be used to pursue the following aims: Aim 1 of the proposal will test a prediction of the hypothesis that changes in the composition of seminal fluid fatty acids and other lipids accompany aging-related loss of fertility. Using multiple mass spectrometry methods, the seminal fluid lipidome will be characterized in young and old males in order to identify features of the lipid profile that correlate with high and low fertility. Aim 2 will test a second prediction of the hypothesis, that the expression of fatty acid synthesis genes in the EB changes in aging males, leading to a shift in fatty acid and related lipid composition in the seminal fluid. Transcriptomic analysis of the EB from young and old males followed by functional validation with genetic knockdown will identify genes that underlie age-related changes in seminal fluid lipid content. In Aim 3, a direct test of the hypothesis will be performed by assessing whether male fertility improves or declines upon genetic manipulation of the seminal fluid fatty acid content. The expected outcomes are significant because they are likely to identify new mechanisms that modulate male and female reproductive physiology and may point to previously unrecognized causes of idiopathic male infertility. The proposed study is innovative because it is expected to identify a new class of molecules and novel biochemical pathways that could be used to improve the diagnosis of infertility, enhance assisted reproductive technology, and treat aging-related decline in fertility.

项目概要/摘要 在许多动物中，精液在雄性生殖成功中发挥着重要作用。精液中的蛋白质 对男性和女性生殖生理的多种特征有显着影响。但相对较少 众所周知，精液的第二个主要成分是脂质，除了其作为精子膜的作用之外 选民。该提案的总体目标是 i) 确定脂质组学和转录组学的差异 年轻和老年男性精液的概况； ii) 直接测试如何操纵精液的脂质谱 液体影响男性生殖成功。该提案将解决精液脂质组的假设 影响男性生育能力并随年龄变化，从而导致与衰老相关的生殖能力下降 成功。这一假设得到了申请人发表的研究的支持，该研究表明脂肪酸和相关的 雄性果蝇精液中的脂质对于雄性生育能力至关重要。两种转座子介导的基因敲除 RNAi 敲低男性射精球 (EB) 中的一种关键脂肪酸生物合成酶，该器官是 不参与精子发生，导致男性生育能力显着下降，强烈支持 精液脂肪酸对男性生殖成功的贡献。与哺乳动物模型相比，使用 果蝇允许在不干扰 EB 的情况下操纵 EB 中产生的精液脂质 睾丸中的精子发生。与脂质组学和转录组学相结合的强大遗传操作 分析将用于实现以下目标： 该提案的目标 1 将测试假设的预测 精液脂肪酸和其他脂质成分的变化伴随着与衰老相关的 生育能力。使用多种质谱方法，将表征年轻和成年患者的精液脂质组。 老年男性，以确定与高生育力和低生育力相关的血脂特征。目标 2 将测试 第二个预测假设，即EB中脂肪酸合成基因的表达随衰老而变化 男性，导致精液中脂肪酸和相关脂质成分的变化。转录组分析 来自年轻和年老男性的 EB，随后通过基因敲除进行功能验证，将识别基因 这是精液脂质含量与年龄相关的变化的基础。在目标 3 中，对假设的直接检验将是 通过评估男性生育能力在精液基因操作后是否提高或下降来进行 液体脂肪酸含量。预期结果很重要，因为它们可能会发现新的 调节男性和女性生殖生理的机制，可能指向以前未被认识到的 特发性男性不育症的原因。拟议的研究具有创新性，因为预计会发现一种新的 可用于改善不孕症诊断的一类分子和新的生化途径， 加强辅助生殖技术，治疗与衰老相关的生育能力下降。