Oocyte development in Drosophila

果蝇卵母细胞的发育

• 批准号: 8837022
• 负责人: Lynn COOLEY
• 金额: $ 61.62万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-07-01 至 2017-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8837022
• 关键词: Adult; Affect; Animals; Apoptosis; Biological; Biological Models; Biology; Cell physiology; Cells; Communication; Cytokinesis; Cytoplasm; Cytoskeleton; Data; Development; Drosophila genus; Drosophila melanogaster; Egg Shell; Egg Yolk Proteins; Ensure; Epithelium; Excision; Exposure to; Food; Funding; Gametogenesis; Genetic; Genetic Recombination; Genetic Transcription; Germ Cells; Giant Cells; Goals; Growth; Health; Hemolymph; Insulin; Insulin Signaling Pathway; Kinesin; Knowledge; Maintenance; Measures; Metabolic; Metabolism; Methods; Microtubules; Modeling; Monitor; Movement; Mus; Nurses; Nutrient; Nutritional; Oocytes; Oogenesis; Ovary; Pathway interactions; Pattern; Peptides; Pharmacologic Substance; Phenotype; Play; Production; Proteins; RNA Interference; Recovery; Research; Role; Shapes; Signal Transduction; Somatic Cell; Spermatocytes; Staging; Starvation; Stem cells; Stress; System; Techniques; Testing; Time; Tissues; Vitellogenesis; base; cost; egg; fly; genetic approach; in vivo; knock-down; mature animal; mutant; novel strategies; nutrition; overexpression; particle; research study; response; sperm cell; tool

DESCRIPTION (provided by applicant): The long-term goal of this project is to elucidate mechanisms of gamete development using Drosophila oogenesis as a model system, and the powerful tools available for genetic and cell biological approaches in Drosophila. A significant health concern is the survival of germline cells in adults exposed to environmental stresses such as poor nutrition and exposure to aggressive pharmaceuticals. Research carried out in this project will impact understanding of protective mechanisms used by immature gametes to ensure their survival through harsh conditions. This project will also investigate a fundamental and conserved aspect of gamete development - some or all of animal gametogenesis occurs in syncytial clusters of cells connected by intercellular bridges called ring canals. Mechanisms modulating intercellular movement of cytoplasm through germline ring canals in response to nutrient availability will be examined. Ring canals are also present between somatic follicle cells of the Drosophila egg chamber, although their function is not known. This project will investigate movement of cytoplasmic components through somatic ring canals, and use novel approaches for disrupting somatic ring canals to determine their function. This information will impact understanding of intercellular movement through small ring canals similar to those connecting developing spermatocytes. In addition, tools developed in this research will be useful for examining somatic ring canals in other Drosophila tissues. Aim 1 will use genetic approaches to determine the mechanism of the starvation response in oocyte development, by manipulating components of the Drosophila Insulin/insulin-like (IIS)/Tor pathway. Previtellogenic egg chambers mount a response to poor nutrition that includes enlarged P bodies and reorganization of the microtubules. Multiple components of the IIS/Tor pathway will be expressed or knocked down, individually or in combinations, in either the germline or the follicle cell tissues to characterize the consequences in starved egg chambers. New approaches will allow tracking of egg chambers that have been starved to examine their recovery in detail, and mutants affecting P body assembly will be used determine the consequences of crippling the starvation response. Aim 2 will use new cell marking techniques and fluorescent proteins to illuminate the function of ring canals in the follicle cell epithelium that surrounds the egg chamber. Experiments will determine whether ring canals serve to equilibrate protein levels between transcriptionally uncoordinated cells. New methods for targeted disruption of somatic ring canals will allow phenotypic analysis of disrupted syncytia, which have been elusive. The importance of the role that follicle cell syncytia play during egg chamber development will be better defined following these studies, and these new methods can then be applied to other somatic syncytia involved in development in Drosophila.

描述（由申请人提供）：该项目的长期目标是以果蝇卵子发生为模型系统阐明配子发育的机制，以及可用于果蝇遗传和细胞生物学方法的强大工具。一个重要的健康问题是暴露于环境压力（如营养不良和暴露于攻击性药物）的成年人生殖细胞的存活。在这个项目中进行的研究将影响对未成熟配子所使用的保护机制的理解，以确保它们在恶劣条件下的生存。该项目还将研究配子发育的一个基本和保守的方面-部分或全部动物配子发生在由细胞间桥连接的合胞体细胞簇中，称为环形通道。调节细胞质通过生殖细胞环道响应营养供应的细胞间运动的机制将被检查。环管也存在于体细胞滤泡细胞之间 果蝇卵室，虽然它们的功能尚不清楚。本项目将研究细胞质成分通过体细胞环管的运动，并使用新的方法破坏体细胞环管，以确定其功能。这一信息将影响理解细胞间运动通过小环管道类似于那些连接发展中的精母细胞。此外，在这项研究中开发的工具将是有用的检查体细胞环管在其他果蝇组织。 目的1将利用遗传学方法，通过操纵果蝇胰岛素/胰岛素样（IIS）/Tor通路的组分，确定卵母细胞发育中饥饿反应的机制。Previtellogenic卵室安装响应营养不良，包括扩大P机构和重组的微管。IIS/Tor途径的多个组分将在种系或卵泡细胞组织中单独或组合表达或敲低，以表征饥饿卵室中的结果。新的方法将允许跟踪已经饥饿的卵室，以详细检查它们的恢复情况，并将使用影响P体组装的突变体来确定削弱饥饿反应的后果。 目的2将使用新的细胞标记技术和荧光蛋白来阐明围绕卵腔的卵泡细胞上皮中的环形通道的功能。实验将确定环形通道是否用于平衡转录不协调细胞之间的蛋白质水平。新的方法有针对性地破坏体细胞环管将允许表型分析破坏合胞体，这一直是难以捉摸的。卵泡细胞合胞体在卵室发育过程中所起作用的重要性将在这些研究之后得到更好的定义，这些新方法可以应用于果蝇发育中涉及的其他体细胞合胞体。