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：
10542735
负责人：
Philip W Jordan
金额：
$ 34.22万
依托单位：
HENRY M. JACKSON FDN FOR THE ADV MIL/MED
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 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10542735
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 Heterozygote Homologous Gene Human Hyperactivity Infertility Knockout Mice 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.

项目摘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划分期间的双极纺锤体确保准确的染色体分离。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性。双极MTOC形成所需的过程是性二态的。染色体精子发生过程中的分离是由含有中心元素的MTOC介导的，这些中心元素在减数分裂I和减数分裂II之前。相反，卵母细胞形成多个acentriolar mToc片段合并在一起形成双极纺锤体。我们已经开发了新的研究工具和改编的新技术来定义和比较MTOC 哺乳动物的精子发生和卵子发生之间的过程。目标1我们的提案重点是确定中心复制的主要调节剂。 polo样激酶4（PLK4）被称为Centriole的“主调节器” 有丝分裂细胞的复制。此外，SAS4是中心位的关键组成部分。但是，很少知道关于在配子发生过程中控制中心重复的控制。在AIM 1A中，我们使用有条件的淘汰鼠标模型有助于确定精子细胞中心重复期间PLK4和SAS4的要求。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AIM 1B中将发现新型的PLK4相互作用伙伴和磷酸化目标，这对于临时性至关重要精子发生过程中中心重复的调节。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的过程大多是不确定的。在AIM 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激酶均已证明可以调节有丝分裂分裂细胞中的集中成熟以重叠的方式。但是，它们在减数分裂中的作用，尤其是与中心体生物发生有关的尚未阐明。在AIM 2A中，我们将使用条件敲除鼠标模型来发现配子发生过程中PLK1和Aurora A激酶，重点放在MTOC过程和染色体上隔离。在AIM 2B中，我们将确定PLK1如何调节Aurora A激酶活性以避免中心过多的。我们还将发现新型生殖细胞特异性中心成分。 PLK1和Aurora A激酶，以确保熟练的中心体生物发生。通过定义哺乳动物期间中心体和acentriolar MTOC生物发生所需的新过程减数分裂我们将开发新的概念，即如何调节减数分裂染色体动力学和隔离。我们提出的研究将有助于配子的诊断性原因，并有助于减少努力这些导致先天缺陷的事件，影响身体和智力发展，并增加不孕症的风险。