Genetics and Proteomics of Mouse Egg Activation

小鼠卵子激活的遗传学和蛋白质组学

• 批准号: 10366090
• 负责人: John C Schimenti
• 金额: $ 23.46万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-15 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10366090
• 关键词: Affect; Animals; Assisted Reproductive Technology; Biochemical; Biological Markers; Biological Models; Calcium; Calcium Signaling; Cell Cycle; Cell Differentiation process; Cell Nucleus; Cells; Chemicals; Data; Deposition; Detection; Development; Diagnosis; Disease; Drosophila genus; Embryo; Embryonic Development; Enzymes; Event; Failure; Fertility; Fertilization; Fertilization in Vitro; Future; Gene Expression; Gene Proteins; Genetic; Genetic Transcription; Genetic study; Genome; Human; Infertility; Invertebrates; Ionophores; Mammals; Maternal Messenger RNA; Mediating; Meiosis; Membrane; Metaphase; Methodology; Modeling; Modification; Molecular; Molecular Genetics; Monitor; Mus; Nature; Nucleic Acids; Oocytes; Oogenesis; Organism; Pathway interactions; Phosphorylation; Phosphotransferases; Placenta; Post-Translational Protein Processing; Procedures; Process; Production; Protein Isoforms; Proteins; Proteome; Proteomics; Rana; Reporting; Role; Sea Urchins; Sterility; Strontium; Testing; Totipotent; Translations; analytical tool; base; calmodulin-dependent protein kinase II; diagnostic tool; egg; female fertility; macromolecule; men; phosphoproteomics; protein expression; sperm cell; tool; transcriptome; translation factor; zygote

PROJECT SUMMARY/ABSTRACT Ovulated human and mouse oocytes are stalled in meiosis II. They are transcriptionally quiescent but have a maternally-loaded transcriptome and proteome. Fertilization triggers “egg activation”, in which a rise(s) of calcium in the oocyte induces several key events that allow transition to embryonic development, namely, meiotic resumption and completion, changes to the egg’s proteome, and genome activation after the first zygotic division. Therefore, egg activation is required for the oocyte to become a totipotent zygote. Despite the essential nature of this process for female fertility, the molecular events of egg activation are not well understood, primarily for technical reasons. Egg activation occurs without new transcription; thus nucleic acids- based ‘omics comparisons are uninformative. The macromolecules that transduce the calcium signal to effect downstream cellular events are not known in humans or any other mammal. Recent studies in MW’s lab exploited technical advantages of the Drosophila model system to show that there is large phospho-modulation of the maternally-provided proteome during egg activation (this also occurs in frogs and sea urchins). We hypothesized, and our genetic data supported, that this posttranslational modification regulated the activity of stored proteins to permit transition of an arrested mature oocyte to a cell that can undertake embryogenesis. We then showed that a calcium-regulated phospho-regulatory enzyme mediates these phospho-changes in the cell cycle machinery, translation factors and other proteins needed to transition the egg to an embryo. In this R21 we propose to test this model for mammalian oocytes, using mouse as a model. Following procedures analogous to those used for Drosophila, we will determine whether there are phosphoproteome changes during mouse egg activation, and which proteins undergo these changes. We will then test the role of CamKII, a calcium-regulated kinase that has been shown to be required for egg activation in mouse, in making these phospho-changes. The results of our studies will lay the groundwork for the field in several ways, including developing phosphoproteomics for mouse oocytes and determining proteins that are phospho-regulated during egg activation. The results will provide information essential for future studies into the roles of the regulated proteins that we identify here, and the effects of specific phosphomodulations during this critical developmental transition. Such fundamental studies will be important for identifying the molecular and genetic bases of human infertilities associated with defective egg activation, providing biomarkers to monitor this process, and potentially for optimizing conditions for assisted reproductive technologies.

项目概要/摘要 排卵的人类和小鼠卵母细胞在减数分裂 II 中停滞，它们在转录上处于静止状态，但已发生减数分裂。 母体负载的转录组和蛋白质组会触发“卵子激活”，其中 卵母细胞中的钙会诱导几个关键事件，从而过渡到胚胎发育，即 减数分裂的恢复和完成、卵子蛋白质组的变化以及第一次减数分裂后的基因组激活 因此，卵母细胞成为全能合子需要卵子激活。 尽管这一过程对于女性生育能力至关重要，但卵子激活的分子事件尚不清楚 理解，主要是由于技术原因，卵子的激活无需新的转录；因此 基于“组学”的比较无法提供传导钙信号的大分子信息。 下游细胞事件在人类或任何其他哺乳动物中尚不清楚。 MW 实验室最近的研究利用果蝇模型系统的技术优势表明： 在卵子激活过程中，母体提供的蛋白质组存在大量磷酸化调节（这种情况也会发生 在青蛙和海胆中）我们勇敢地认为，并且我们的遗传数据支持这种翻译后过程。 修饰调节储存蛋白质的活性，以允许停滞的成熟卵母细胞转变为细胞 然后我们发现了一种钙调节的磷酸调节酶。 介导细胞周期机制、翻译因子和其他蛋白质的磷酸化变化 将卵子转变为胚胎。 在此 R21 中，我们建议使用小鼠作为模型来测试哺乳动物卵母细胞的模型。 类似于用于果蝇的程序，我们将确定是否存在磷酸化蛋白质组 小鼠卵子激活过程中的变化，以及哪些蛋白质经历这些变化，然后我们将测试其作用。 CamKII 是一种钙调节激酶，已被证明是小鼠卵子激活所必需的，在 这些磷酸化变化。 我们的研究结果将从多个方面为该领域奠定基础，包括开发 小鼠卵母细胞的磷酸蛋白质组学和确定卵子过程中磷酸调节的蛋白质 结果将为未来研究受监管的作用提供重要信息。 我们在这里确定的蛋白质，以及在这个关键发育过程中特定磷酸调节的影响 此类基础研究对于确定人类的分子和遗传基础非常重要。 与卵子激活缺陷相关的不孕症，提供生物标志物来监测这一过程，以及 潜在地优化辅助生殖技术的条件。

项目成果

Cilia take the egg on a magic carpet ride.

纤毛带着鸡蛋乘坐魔毯。

• DOI: 10.1073/pnas.2108887118
• 发表时间: 2021
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Suarez,SusanS;Wolfner,MarianaF
• 通讯作者: Wolfner,MarianaF