HORMONE ACTION IN THE CENTRAL NERVOUS SYSTEM

中枢神经系统中的激素作用

• 批准号: 3080921
• 负责人: ALAN P FARWELL
• 金额: $ 7.66万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 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 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3080921
• 关键词: actins; affinity chromatography; autoradiography; axon; binding proteins; cell differentiation; cell free system; cell growth regulation; central nervous system; cytoskeleton; electron microscopy; gel electrophoresis; hormone regulation /control mechanism; immunocytochemistry; laboratory rat; radiotracer; thyroid hormones; thyroxine; tissue /cell culture

The goal of this project is the characterization of the mechanism(s) by which thyroid hormone regulates actin polymerization and, thus, affects the integrity of the actin cytoskeleton in cells of the Central Nervous System. Because alterations in polymerization of the actin cytoskeleton would significantly affects the complex interactions between the cytoskeletal network and the extracellular matrix, resulting in attenuation of neuronal growth and development, studies of the interactions of T4 with the actin cytoskeleton will provide insight into thyroid hormone's role in the growth and differentiation of neurons. The model system to be examined is the T4- dependent regulation of type II iodothyronine 5'-deiodinase (5'D-II) in cultured, cAMP-stimulated glial cells. T4 modulates levels of this important cerebrocortical enzyme by decreasing the biological half-life of 5'D-II through a novel, extra-nuclear mode of action. Disruption of the microfilaments blocks this T4-mediated action, indicating that the actin cytoskeleton is a likely site for regulation by thyroid hormone. 5'D-II can be affinity labeled by a T4 derivative and, thus, can be visualized in the cell using immunocytochemical and histochemical techniques. The morphological characterization of the interactions between T4, actin and 5'D-II will be accomplished during the first phase of this project. The second phase of the project will involve the identification of the mediator(s) of thyroid hormone's effects on the cytoskeleton. Actin and T4 affinity chromatography will be used to isolate potential actin- binding proteins which will then be evaluated in a cell free system to asses their ability to affect actin polymerization. Protein(s) that bind to filaments actin and T4 and that demonstrate a T4-modulated effect on actin polymerization will be characterized further with 2D SDS-PAGE and peptide mapping and compared to information available on actin-binding proteins previously described. The identification and characterization of the mechanism(s) by which T4 interacts with the cytoskeleton, and thereby regulates the turnover of a short-lived membrane protein, will then allow the study of the more fundamental questions regarding thyroid hormone's influence on, and participation in, the growth of developing neurons and the subsequent establishment of cell-cell and cell-substratum attachments.

该项目的目标是通过以下方式对机制进行定性： 甲状腺激素调节肌动蛋白聚合，从而影响 中枢神经系统细胞中肌动蛋白细胞骨架的完整性 系统 因为肌动蛋白细胞骨架聚合的改变 会显著影响 细胞骨架网络和细胞外基质，导致 神经生长和发育的衰减， T4与肌动蛋白细胞骨架的相互作用将提供深入了解 甲状腺激素在神经元生长和分化中的作用。 的 要检查的模型系统是T4依赖的II型调节 培养的cAMP刺激的神经胶质细胞中的碘甲腺原氨酸5 '-脱碘酶（5' D-II） 细胞 T4通过以下方式调节这种重要的肾上腺皮质酶的水平： 通过一种新的， 核外作用模式。 微丝阻滞的破坏 这种T4介导的作用，表明肌动蛋白细胞骨架是一种 可能是受甲状腺激素调节的部位。 5 ′ D-II可以是亲和的 标记的T4衍生物，因此，可以在细胞中可视化， 免疫细胞化学和组织化学技术。 形态 T4，肌动蛋白和5 'D-II之间的相互作用的表征将是 在这个项目的第一阶段完成。 第二阶段 该项目将涉及确定甲状腺激素的介体， 激素对细胞骨架的影响。 肌动蛋白和T4亲和力 层析将被用来分离潜在的肌动蛋白结合蛋白 然后将在无细胞系统中进行评估， 影响肌动蛋白聚合。与肌动蛋白丝结合的蛋白质 和T4，证明了T4对肌动蛋白聚合的调节作用 将用2D SDS-PAGE和肽图谱进一步表征， 与之前关于肌动蛋白结合蛋白的信息相比， 介绍了 通过以下方式确定和表征机制： 其中T4与细胞骨架相互作用，从而调节 一种短寿命的膜蛋白的周转，然后将允许研究 关于甲状腺激素的影响， 参与神经元的生长以及随后的 细胞-细胞和细胞-基质附着的建立。