EGG MATURATION: HORMONE RECEPTOR SITES AND UPTAKE

卵子成熟：激素受体位点和吸收

• 批准号: 3318292
• 负责人: THOMAS E SCHROEDER
• 金额: $ 11.08万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-09-30 至 1986-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3318292
• 关键词: adenine analog; autoradiography; cell cycle; chemical binding; cytoskeleton; egg /ovum; electron microscopy; fluorescence microscopy; hormone metabolism; membrane activity; microscopy; morphology; oogenesis; pinocytosis; receptor mediated endocytosis; saltwater environment; scintillation counter; videotape /videodisc

This project aims to investigate the early interactions between a defined hormone (l-methyladenine = 1-MA) and its target cell (starfish oocyte) that result in dramatic changes of the cytoskeleton and stimulate cell division. It is known that l-MA initially binds at the cell-surface of the oocyte and is transduced to they cytoplasma where is stimulates the completion of maturation divisions. This hormone-oocyte interaction, therefore, represents an important model system for studying hormone-cell surface interactions in general, as well as similar hormone-oocyte interactions in vertebrates including human, and the regulation of cell division. This project represents one of the Principal Investigator's long-term objectives: elucidation of the mechanisms of cell division. Experiments will test specific hypotheses concerning the mophological distribution of l-MA receptor sites on the oocyte surface and the cellular mechanism of l-MA uptake. They will employ molecular probes and markers in a set of morphological analyses. Tritiated-1-MA will be used to localize the initial binding sites of l-MA and reveal its intracellular pathway by autoradiography and light and electron microscopy. Fluid-content markers will be used to explore the existence, pathway and kinetics of endocytosis as the mechanism of hormone uptake in oocytes before and after application of l-MA; these agents include a fluorescent market (Lucifer Yellow CH) and a cytochemical marker (horseradish peroxidase) for microscopic analyses, and will be complemented by kinetic analyses of tritiated-insulin uptake using scintillation counting. These studies will reveal th role of receptor-mediated endocytosis in hormone-oocyte interations. Normal processes will be manipulated with two kinds of inhibitory drugs: antagonists of oocyte maturation and inhibitors of cytoskeletal organization and function. These manipulations will test the roles of the actin- and tubulin-based cytoskeletons in hormone transductiion and will explore the subcellular actions of known antagonists of oocyte maturation.

这个项目旨在研究一个定义的 激素（1-甲基腺嘌呤= 1-MA）及其靶细胞（海星卵母细胞）， 导致细胞骨架的剧烈变化，刺激细胞 师. 已知I-MA最初结合在肿瘤细胞的细胞表面， 卵母细胞，并被转导到他们的细胞质，在那里刺激 成熟分裂的完成。 这种胚胎-卵母细胞的相互作用， 因此，它是一个重要的模型系统， 一般来说，表面相互作用以及类似的受精卵-卵母细胞 包括人类在内的脊椎动物之间的相互作用，以及细胞 师. 该项目代表了主要研究者之一 长期目标：阐明细胞分裂的机制。 实验将检验有关形态学的特定假设。 卵母细胞表面和细胞膜上l-MA受体位点的分布 L-MA摄取机制。 他们将使用分子探针和标记， 一组形态学分析 氚化-1-MA将用于定位 的初始结合位点，并揭示其细胞内途径， 放射自显影和光学和电子显微镜。 液体含量标记物 将用于探索内吞作用的存在、途径和动力学 作为应用前后卵母细胞中激素摄取的机制 这些试剂包括荧光试剂（Lucifer Yellow CH）和 用于显微镜分析的细胞化学标记物（辣根过氧化物酶）， 并将通过氚化胰岛素摄取的动力学分析进行补充 使用闪烁计数。 这些研究将揭示 受体介导的内吞作用。 正常的过程将被两种抑制药物操纵： 卵母细胞成熟的拮抗剂和细胞骨架的抑制剂 组织和功能。 这些操作将测试 肌动蛋白和微管蛋白为基础的细胞骨架在激素转导和意志 探索已知卵母细胞成熟拮抗剂的亚细胞作用。