Administrative Supplement request for GM117155

GM117155 行政补充申请

• 批准号: 10790343
• 负责人: Philip W Jordan
• 金额: $ 10.62万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 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/10790343
• 关键词: Address; Administrative Supplement; Affect; Aneuploidy; Biogenesis; Cell Cycle; Centrioles; Centrosome; Chemicals; Chromosome Segregation; Chromosome abnormality; Chromosomes; Clinical; Congenital Abnormality; Cryoultramicrotomy; DNA biosynthesis; Development; Diagnosis; Down Syndrome; Ensure; Equipment; Event; Failure; Fertilization; Funding; Gametogenesis; Genetic; Genetic Diseases; Germ Cells; Goals; Gonadal structure; Haploidy; Homologous Gene; Human; Infertility; Knowledge; Marketing; Mediating; Meiosis; Methods; Microtubule-Organizing Center; Modeling; Mus; Mutation; Oocytes; Oogenesis; Organ Procurements; Ovary; Phosphotransferases; Pregnancy; Pregnancy loss; Process; Protocols documentation; Regulation; Reporting; Research; Resolution; Risk; Sampling; Sister Chromatid; Spermatocytes; Spermatogenesis; System; Techniques; Testis; Tissue Expansion; cryostat; egg; experimental study; improved; mental development; novel; prevent; repaired; segregation; sexual dimorphism; small molecule inhibitor; sperm cell; superresolution microscopy; theories; tool

PROJECT SUMMARY The establishment of bipolar spindles during meiotic divisions ensures accurate chromosome segregation. Characterization of microtubule organizing center (MTOC) dynamics will help understand the 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. In Aim 1, we will assess meiotic progression in the absence of key microtubule organizing center (MTOC) regulators. In Aim 2, we aim to discover and characterize novel components of MTOCs during oogenesis and spermatogenesis. In order to achieve our objectives, we have developed a method to increase the magnification of spermatocytes and oocytes by combining cryosectioning with tissue expansion and super-resolution microscopy to enable us to view fine structural features of MTOCs. In addition, with donated, deidentified human gonad samples that we receive via approved protocols with organ procurement organizations, we are performing short-term culture experiments and chemical modulation using specific small molecule inhibitors against cell cycle kinases. Our aims can only be addressed with the appropriate equipment. Unfortunately, our cryostat recently broke down without an option for repair (discontinued model). Therefore, we are requesting a new, improved cryostat system to alleviate this issue. The Epredia Cryostar NX70 we propose to obtain with equipment supplement funding is optimal for our needs, a superior product, and will solve our current cryosectioning issue. 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 在减数分裂期间双极纺锤体的建立确保了染色体的精确分离。 微管组织中心（MTOC）动力学的表征将有助于理解配子的原因 非整倍性形成双极MTOC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粒的MTOC介导的，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。相反，卵母细胞形成多个无中心MTOC片段， 结合在一起形成双极纺锤体。 我们已经开发了新的研究工具和采用新的技术来定义和比较MTOC 哺乳动物精子发生和卵子发生之间的过程。在目标1中，我们将评估减数分裂进程 在缺乏关键微管组织中心（MTOC）调节剂的情况下。在目标2中，我们旨在发现和 表征卵子发生和精子发生过程中MTOC的新组分。为了实现我们 目的，我们已经开发了一种方法，以增加放大的精母细胞和卵母细胞， 结合冷冻切片、组织扩张和超分辨率显微镜， MTOC的结构特征。此外，通过捐赠的，去识别的人类性腺样本，我们通过 与器官采购组织批准的协议，我们正在进行短期培养实验 和使用针对细胞周期激酶的特异性小分子抑制剂的化学调节。 我们的目标只能通过适当的设备来实现。不幸的是我们的低温恒温器最近坏了 没有维修选项（停产型号）。因此，我们需要一种新的、改进的低温恒温器 系统来缓解这个问题。我们建议通过设备补充获得的Epredia Cryostar NX70 资金是最适合我们的需求，一个上级产品，并将解决我们目前的冷冻切片问题。 通过定义哺乳动物中中心体和无中心粒MTOC生物发生所需的新过程， 减数分裂我们将发展减数分裂染色体动力学和分离是如何调节的新概念。 我们提出的研究将有助于诊断配子非整倍体的原因，并有助于减少 这些事件导致出生缺陷，影响身心发育，增加不孕风险。