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Regulation of microtubule organizing centers during mammalian gametogenesis

哺乳动物配子发生过程中微管组织中心的调节

基本信息

批准号：
10321962
负责人：
Philip W Jordan
金额：
$ 28.83万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-01-01 至 2022-05-20
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10321962
关键词：
Address Affect Aneuploidy Biogenesis Bispecific Antibody 2B1 Cell Cycle Cells Centrioles Centrosome Chromosome Pairing Chromosome Segregation Chromosome abnormality Chromosomes Clinical Congenital Abnormality DNA biosynthesis Data Development Diagnosis Ensure Event Excision Fertility Fertilization Gametogenesis Genetic Diseases Genetic Recombination Germ Cells Haploidy Human Hyperactivity Infertility Knowledge Mediating Meiosis Microtubule-Organizing Center Mitotic Modeling Modification Mus Mutation Oocytes Oogenesis PLK1 gene Phase Transition Phosphorylation Phosphotransferases Pregnancy Procentriole Process Property Proteins Regulation Reporting Research Risk Role S phase Series Sister Chromatid Spermatocytes Spermatogenesis System Techniques Testing Yeasts aurora kinase A conditional knockout egg mental development mouse model next generation novel prevent segregation sexual dimorphism sperm cell tool yeast two hybrid system

项目摘要

PROJECT SUMMARY Regulation of microtubule organizing centers during mammalian gametogenesis Establishment of bipolar spindles during meiotic divisions ensures accurate chromosome segregation. Characterization of microtubule organizing center (MTOC) dynamics will help understand causes of gamete aneuploidy. The processes required for the formation of bipolar MTOCs are sexually dimorphic. Chromosome segregation during spermatogenesis is mediated by MTOCs containing centrioles that duplicate once prior to meiosis I and again prior to meiosis II. In contrast, oocytes form multiple acentriolar MTOC fragments that coalesce together to form bipolar spindles. We have developed new research tools and adapted novel techniques to define and compare MTOC processes between mammalian spermatogenesis and oogenesis. Aim 1 of our proposal focuses on determining key regulators of centriole duplication. Polo-like kinase 4 (PLK4) is known as the “master regulator” of centriole duplication in mitotic cells. In addition, SAS4 is a key component of the centriole. However, very little is known about the control of centriole duplication during gametogenesis. In Aim 1A, we use conditional knockout mouse models to help determine the requirements for PLK4 and SAS4 during centriole duplication in spermatocytes. Furthermore, we will use these models to address whether they have functions during oogenesis. In Aim 1B we will discover the novel PLK4 interaction partners and phosphorylation targets that are critical for temporal regulation of centriole duplication during spermatogenesis. The processes of centrosome maturation and separation required for bipolar spindle formation during meiosis are mostly undefined. In Aim 2 we will assess key stages of centrosome biogenesis during spermatogenesis. PLK1 and Aurora A kinases have both been shown to regulate centrosome maturation in mitotically dividing cells in an overlapping manner. However, their roles during meiosis, particularly relating to centrosome biogenesis, are yet to be elucidated. In Aim 2A, we will use conditional knockout mouse models to discover the functions of PLK1 and Aurora A kinases during gametogenesis, with focus on MTOC processes and chromosome segregation. In Aim 2B we will determine how PLK1 regulates Aurora A kinase activity to avoid centriole overduplication. We will also discover novel germ cell specific centrosomal components that are targeted by PLK1 and Aurora A kinases to ensure proficient centrosome biogenesis. By defining the novel processes required for centrosome and acentriolar MTOC biogenesis during mammalian meiosis we will develop new concepts of how meiotic chromosome dynamics and segregation are regulated. Our proposed research will contribute to diagnosing causes of gamete aneuploidy and help with efforts to reduce these events that cause birth defects, affect physical and mental development, and increase the risk of infertility.

项目概要哺乳动物配子发生过程中微管组织中心的调节减数分裂期间双极纺锤体的建立确保了准确的染色体分离。微管组织中心 (MTOC) 动力学的表征将有助于了解配子的原因非整倍体。双极 MTOC 形成所需的过程是两性二态的。染色体精子发生过程中的分离是由含有中心粒的 MTOC 介导的，该中心粒在精子发生前复制一次减数分裂 I 和减数分裂 II 之前再次进行。相反，卵母细胞形成多个无心粒 MTOC 片段，结合在一起形成双极纺锤体。我们开发了新的研究工具并采用了新技术来定义和比较 MTOC 哺乳动物精子发生和卵子发生之间的过程。我们提案的目标 1 重点是确定中心粒复制的关键调节因子。 Polo样激酶4（PLK4）被称为中心粒的“主调节器” 有丝分裂细胞中的重复。此外，SAS4是中心粒的关键组成部分。然而，人们知之甚少关于配子发生过程中中心粒复制的控制。在目标1A中，我们使用条件敲除小鼠模型可帮助确定精母细胞中心粒复制期间对 PLK4 和 SAS4 的需求。此外，我们将使用这些模型来解决它们在卵子发生过程中是否具有功能。在目标 1B 中，我们将发现对时间至关重要的新型 PLK4 相互作用伙伴和磷酸化靶点精子发生过程中中心粒复制的调节。减数分裂过程中双极纺锤体形成所需的中心体成熟和分离过程大多是未定义的。在目标 2 中，我们将评估精子发生过程中中心体生物发生的关键阶段。 PLK1 和 Aurora A 激酶均已被证明可以调节有丝分裂细胞的中心体成熟以重叠的方式。然而，它们在减数分裂过程中的作用，特别是与中心体生物发生相关的作用，尚待阐明。在Aim 2A中，我们将使用条件敲除小鼠模型来发现配子发生过程中的 PLK1 和 Aurora A 激酶，重点关注 MTOC 过程和染色体隔离。在目标 2B 中，我们将确定 PLK1 如何调节 Aurora A 激酶活性以避免中心粒过度重复。我们还将发现新的生殖细胞特异性中心体成分，这些成分是 PLK1 和 Aurora A 激酶可确保熟练的中心体生物发生。通过定义哺乳动物过程中中心体和无心粒 MTOC 生物发生所需的新过程减数分裂我们将开发如何调节减数分裂染色体动力学和分离的新概念。我们提出的研究将有助于诊断配子非整倍体的原因，并有助于努力减少配子非整倍体的发生。这些事件会导致出生缺陷，影响身心发育，并增加不孕的风险。