EGG MATURATION: HORMONE RECEPTOR SITES AND UPTAKE

该项目旨在调查已定义的 激素(L-甲基腺嘌呤=1-MA)及其靶细胞(海星卵母细胞) 导致细胞骨架发生剧烈变化，刺激细胞 组织。已知L-MA最初结合在细胞表面。 并被转导到细胞质，在那里IS刺激 成熟期划分完成。这种荷尔蒙-卵母细胞相互作用， 因此，它代表了研究激素细胞的重要模型系统。 一般的表面相互作用，以及类似的激素-卵母细胞 包括人类在内的脊椎动物的相互作用和细胞的调节 组织。该项目代表首席调查员之一 长期目标：阐明细胞分裂的机制。 实验将检验有关形态的特定假说 L-MA受体在卵母细胞表面和细胞内的分布 L-MA的摄取机制。他们将使用分子探测器和标记来 一组形态分析。Tritiated-1-MA将用于本地化 L-MA的初始结合部位及其胞内途径 放射自显影和光学和电子显微镜。流体含量标记物 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激素摄取的机制 这些药物包括荧光市场(荧光黄CH)和 一种用于显微镜分析的细胞化学标记(辣根过氧化物酶)， 并将由氚-胰岛素摄取的动力学分析来补充 使用闪烁计数。这些研究将揭示 激素-卵母细胞相互作用中受体介导的内吞作用。 正常的过程将被两种抑制药物操纵： 卵母细胞成熟拮抗剂和细胞骨架抑制剂 组织和职能。这些操作将测试 基于肌动蛋白和微管蛋白的细胞骨架在激素转导和意志中的作用 探索已知的卵母细胞成熟拮抗剂的亚细胞作用。

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.