Oocyte development in Drosophila

果蝇卵母细胞的发育

• 批准号: 8369693
• 负责人: Lynn COOLEY
• 金额: $ 61.43万
• 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 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8369693
• 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; 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. PUBLIC HEALTH RELEVANCE: Eggs and sperm are the ultimate stem cells, providing the starting point for entire adult animals. This project will study fundamental aspects of egg development using the fruit fly Drosophila melanogaster as a model system and the powerful experimental tools available in flies. The research will help our understanding of how immature eggs protect themselves during periods of poor nutrition.

描述（由申请人提供）：该项目的长期目标是利用果蝇卵子发生作为模型系统来阐明配子发育机制，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（例如营养不良和接触侵蚀性药物）的成年人中生殖细胞的存活。该项目中进行的研究将影响对未成熟配子使用的保护机制的理解，以确保其在恶劣条件下生存。该项目还将研究配子发育的一个基本且保守的方面——部分或全部动物配子发生发生在由称为环管的细胞间桥连接的合胞体簇中。将检查通过种系环管调节细胞质的细胞间运动以响应营养物质可用性的机制。体卵泡细胞之间也存在环管 果蝇卵室的结构，尽管其功能尚不清楚。该项目将研究细胞质成分通过体细胞环管的运动，并使用破坏体细胞环管的新方法来确定其功能。这些信息将影响对通过类似于连接发育中的精母细胞的小环管的细胞间运动的理解。此外，本研究开发的工具将有助于检查其他果蝇组织的体细胞环管。 目标 1 将使用遗传方法通过操纵果蝇胰岛素/类胰岛素 (IIS)/Tor 通路的成分来确定卵母细胞发育中饥饿反应的机制。卵黄发生前的卵室对营养不良做出反应，包括 P 体增大和微管重组。 IIS/Tor 通路的多个成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后果。新方法将允许跟踪已经饥饿的卵室，以详细检查其恢复情况，并且将使用影响 P 体组装的突变体来确定削弱饥饿反应的后果。 目标 2 将使用新的细胞标记技术和荧光蛋白来阐明卵室周围的卵泡细胞上皮中环管的功能。实验将确定环管是否有助于平衡转录不协调细胞之间的蛋白质水平。定向破坏体细胞环管的新方法将允许对破坏的合胞体进行表型分析，而这一直是难以捉摸的。这些研究将更好地确定滤泡细胞合胞体在卵室发育过程中发挥的作用的重要性，并且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。 公共健康相关性：卵子和精子是最终的干细胞，为整个成年动物提供了起点。该项目将使用果蝇果蝇作为模型系统以及果蝇中可用的强大实验工具来研究卵发育的基本方面。这项研究将有助于我们了解未成熟的鸡蛋在营养不良期间如何保护自己。