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.

项目摘要/摘要 排卵的人和小鼠卵母细胞在减数分裂II中停滞不前。他们是转录静止的，但是 主要负载的转录组和蛋白质组。受精触发了“卵子激活”，其中升高 卵母细胞中的钙诱导了几个关键事件，这些事件允许过渡到胚胎发育，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激活的基因组激活 合子司。因此，卵母细胞成为全能的合子需要卵激活。尽管有 女性生育过程的基本性质，卵子激活的分子事件不好 理解，主要是出于技术原因。卵激活发生没有新的转录；因此核酸 - 基于“ OMICS比较”是无信息的。转导钙信号的大分子作用 下游细胞事件在人类或任何其他哺乳动物中尚不清楚。 MW实验室的最新研究探讨了果蝇模型系统的技术优势，以表明 在鸡蛋激活过程中，主要提供的蛋白质组有大量的磷酸调节（这也发生 在青蛙和海胆中）。我们假设我们的遗传数据是这种翻译后的 修改调节储存蛋白的活性，以允许被捕的成熟卵母细胞过渡到细胞 可以进行胚胎发生。然后，我们表明钙调节的磷酸化调节酶 介导细胞周期机械中的这些磷酸变化，翻译因子和其他所需的蛋白质 将鸡蛋过渡到胚胎。 在此R21中，我们建议使用小鼠作为模型来测试该模型的哺乳动物卵母细胞。下列的 类似于果蝇类似的程序，我们将确定是否存在磷蛋白酶 小鼠卵激活期间的变化，哪些蛋白质会发生这些变化。然后，我们将测试 CAMKII是一种钙调节的激酶 这些磷酸变化。 我们的研究结果将以几种方式为该领域奠定基础，包括开发 小鼠卵母细胞的磷酸蛋白质组学，并确定卵子在卵中受到磷酸化的蛋白质 激活。结果将为未来研究受监管的作用提供必不可少的信息 我们在这里识别的蛋白质以及在这种关键发育过程中特定磷酸化的影响 过渡。这样的基本研究对于识别人类的分子和遗传基础很重要 与有缺陷的卵子激活相关的上等实用性，提供生物标志物来监测此过程，并 有可能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