Comparative analysis of molecular events in mammalian oocytes

哺乳动物卵母细胞分子事件的比较分析

• 批准号: 10755189
• 负责人: AHMED BALBOULA
• 金额: $ 32.5万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-04 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10755189
• 关键词: Ablation; Age; Aneuploidy; Animals; Area; Assisted Reproductive Technology; Behavior; Biological; Biological Assay; Birth; CDC42 gene; Cattle; Cell Nucleus; Cell division; Chromatin; Chromosome Segregation; Chromosomes; Clinical; Competence; Confocal Microscopy; Congenital Abnormality; Cues; Data; Defect; Development; Domestic Animals; Embryo; Embryo Loss; Ensure; Event; Exposure to; Failure; Family suidae; Female; Fertility; Gene Expression; Generations; Genetic; Germ Cells; Goals; Human; Incidence; Infertility; Insurance; Kinetochores; Knowledge; Lasers; Lead; Life; Meiosis; Meiotic Prophase I; Microtubules; Molecular; Molecular Analysis; Mus; National Institute of Child Health and Human Development; Oocytes; Ovulation; Peripheral; Personal Satisfaction; Positioning Attribute; Pregnancy; Process; Puberty; Public Health; Reproductive Health; Research; Risk Factors; Risk Reduction; Rodent; Science; Signal Transduction; Spontaneous abortion; Vesicle; Woman; Work; chromosome missegregation; chromosome number abnormality; comparative; developmental disease; early pregnancy loss; egg; female fertility; fetal; genetic approach; improved; innovation; male; migration; multi-photon; multiple omics; novel; novel strategies; offspring; oocyte maturation; oocyte quality; premature; sperm cell; success; superresolution microscopy; transcriptomics

Project summary Egg generation entails a unique cell division, Meiosis I (a process also known as oocyte maturation), during which the oocyte acquires its developmental competence. For poorly understood reasons, mammalian oocytes (including human, bovine and porcine oocytes) are notoriously prone to reduced quality. Because oocyte quality is a major determinant of embryo developmental competence and pregnancy success, the long-term research goal of this application is to dissect basic molecular mechanisms that regulate meiosis I in human, bovine and porcine oocytes to understand why female gametes are notoriously prone to reduced quality. Work from our group has demonstrated that preferential initial nucleus and spindle positioning at the center of mouse oocytes is an insurance mechanism to avoid the premature exposure of the spindle to the cortical signaling that hinders proper chromosome-microtubule attachments, thereby protecting against aneuploidy, the major genetic cause of infertility and congenital defects. These findings implicate initial peripheral nucleus and spindle positioning as a major risk factor for aneuploidy in mice. Strikingly, in contrast to mouse oocytes, the majority of human, bovine and porcine oocytes have a peripheral nucleus and, subsequently, peripheral spindle formation. Interestingly, in human oocytes, peripheral nucleus positioning correlates with poor maturation rates. How the nucleus/spindle behaves when it is initially positioned peripherally in those species (humans, pigs and cattle) and whether this behavior correlates with aneuploidy are unknown. This proposal builds on these findings via two complementary albeit independent aims: (1) determine the underlying molecular mechanisms regulating nucleus behavior in human, bovine and porcine oocytes, and (2) determine whether nucleus/spindle positioning relates to aneuploidy. To carry out these aims, we will employ super-resolution microscopy, multiphoton laser ablation, genetic, optochemical and integrative multiomic approaches. These conceptually and technically innovative aims are expected to have a broad impact on the field by filling a substantial gap in our knowledge of how the nucleus/spindle behaves in human, bovine and porcine oocytes to ensure the development of good-quality gametes with clear clinical implications for assisted reproductive technologies. Such understanding is relevant to human and animal reproductive health because mammalian oocytes are notoriously associated with poor developmental competence and aneuploidy, major causes of early pregnancy loss (a high priority area of the Fertility and Infertility Branch of the NICHD).

项目摘要 鸡蛋产生需要独特的细胞分裂，减数分裂I（也称为卵母细胞成熟的过程），在此期间，卵母细胞具有其发育能力。出于鲜为人知的原因，哺乳动物卵母细胞（包括人，牛和猪卵母细胞）众所周知，质量降低。由于卵母细胞质量是胚胎发育能力和妊娠成功的主要决定因素，因此该应用的长期研究目标是剖析基本的分子机制，这些基本分子机制可以调节人，牛和猪卵母细胞中的减数分裂I，以了解为什么众所周知，女性配子众所周知，众所周知，女性配子会降低质量降低质量。我们小组的工作表明，在小鼠卵母细胞中心的优先初始核和主轴定位是一种保险机制，旨在避免纺锤体过早暴露于皮质信号中，以阻碍适当的染色体 - 微管附件，从而降低适当的染色体，从而防止动脉粥样硬化。 不育和先天性缺陷。这些发现暗示初始外周核和纺锤体定位是小鼠非整倍性的主要危险因素。令人惊讶的是，与小鼠卵母细胞相比，大多数人，牛和猪卵母细胞具有外周核，随后是外周纺锤体的形成。有趣的是，在 人卵母细胞，外周核定位与成熟率较差有关。当核最初将其外周放在这些物种（人类，猪和牛）中时，该核的表现如何以及这种行为是否与非整倍性相关。 该提案通过两个互补的发现建立在这些发现的基础上：（1）确定调节人，牛和猪卵母细胞中核行为的潜在分子机制，以及（2）确定核/纺锤体的位置是否与舌相关。为了实现这些目标，我们将采用超分辨率显微镜，多光子激光消融，遗传，验光和综合多构方法。这些在概念和技术上创新的目标有望通过在人，牛和猪牛肉卵母细胞中填补核/纺锤体的表现，从而对该领域产生广泛的影响，以确保对辅助生殖技术的清晰临床含义的开发，以确保具有清晰的临床意义。这种理解与人类和动物的生殖健康有关，因为众所周知，哺乳动物卵母细胞与发育能力差和非整倍性相关，这是早期妊娠早期丧失的主要原因（NICHD的生育能力和不育分支的高优先区域）。