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.

项目摘要/摘要 在许多动物中，第二液在男性生殖成功中起着重要作用。第二液中的蛋白质 对男性和女性生殖生理学的多种特征有重大影响。但是，相对较少 已知第二个流体的第二个主要成分，即脂质，超出其作为精子膜的角色 成分。该提案的总体目标是i）确定脂质组和转录组的差异 年轻男性和老年男性第二液的特征； ii）直接测试如何操纵第二个 流体影响男性生殖成功。该提议将解决第二液脂肪组的假设 影响男性生育能力和随着年龄的变化，从而导致与衰老相关的生殖的下降 成功。申请人已发表的研究表明，这一假设表明脂肪酸和相关 果蝇雄性第二液中的脂质对于雄性生育至关重要。两个转座子介导的淘汰赛 雄性射精灯泡（EB）中的钥匙脂肪酸生物合成酶的RNAi敲低是一种器官 不参与精子发生，导致男性生育能力显着降低，强烈支持 半流体脂肪酸对男性生殖成功的贡献。与哺乳动物模型相反，使用 果蝇允许在EB中产生的半流体脂质进行操纵，而不会干扰 测试中的精子发生。强大的遗传操纵与脂质组和转录组结合 分析将用于追求以下目的：该提案的目标1将检验假设的预测 半流体脂肪酸和其他脂质的组成的变化涉及与衰老相关的损失 生育能力。使用多种质谱法，第二液脂肪组将在Young和 老男性以识别与高生育率相关的脂质剖面的特征。 AIM 2将测试 假设的第二个预测，即EB中脂肪酸合成基因的表达在衰老中的变化中 雄性，导致第二液中脂肪酸和相关脂质组成的转移。转录组分析 来自年轻男性和老年男性的EB，然后用遗传敲低的功能验证识别基因 这是半流体脂质含量的与年龄相关的变化。在AIM 3中，对假设的直接检验将是 通过评估男性生育能力在第二次遗传操作后是否改善还是下降 流体脂肪酸含量。预期的结果很重要，因为它们可能会识别新的结果 调节男性和女性生殖生理的机制，可能指出以前未被认可的机制 特发性男性不育症的原因。拟议的研究具有创新性，因为它有望确定一个新的 一类分子和新型的生化途径，可用于改善不育的诊断， 增强辅助生殖技术，并治疗与衰老相关的生育能力下降。