MORPHOLOGICAL STUDIES IN DIABETES AND METABOLISM

这项提议的目标是使用形态和生化 分析胰岛素受体调节的方法。超微结构研究 在胰岛素敏感细胞上显示出显著的细胞特异性 胰岛素的组织、分布和移动性的异质性 感受器。这些形态观察与细胞特异性相关 在相同细胞的生化结合研究中发现了差异。近期 有证据表明，细胞间的可变性可能存在于这些过程中。 参与胰岛素受体内化和再循环。 这项建议的具体目标是： (1)定性、定量测定和比较胰岛素 受体在不同细胞类型上的组织和分布。 如果有关系，将在被占用的胰岛素之间建立关系 受体和特定的细胞表面结构和成分。 (2)分析细胞胰岛素受体占用后的迁移率 它们显示了占用引起的受体组织或 分发。将使用各种化学试剂试图阻止 受体微聚集，以确定所涉及的机制和 与胰岛素作用的关系。 (3)确定涉及的路线和亚细胞结构 胰岛素的内化、降解或加工和再循环 胰岛素受体。 除了使用单体铁蛋白-胰岛素进行形态研究外， 抗胰岛素受体抗体和免疫细胞化学 生物素标记的胰岛素与胰岛素受体结合后产生抗体 或亲和素结合的胶体金粒子和/或过氧化物酶 用来显示完整的胰岛素受体以及 通透性细胞。这个实验室处于一个独特的位置，可以完成 这一建议是因为我们被证明有能力执行量化 形态及相关生化研究。这些研究将 有助于我们理解胰岛素的作用和可能的作用部位 胰岛素抵抗状态下的结合后缺陷。

The goal of this proposal is to use morphological and biochemical approaches to analyze insulin receptor regulation. Ultrastructural studies on insulin-sensitive cells have shown significant cell-specific heterogeneity in the organization, distribution and mobility of insulin receptors. These morphological observations correlate with cell-specific differences found in biochemical binding studies on the same cells. Recent evidence suggests that cell-to-cell variability may exist in the processes involved in insulin receptor internalization and recycling. The specific aims of this proposal are: (1) to qualitatively and quantitatively determine and compare insulin receptor organization and distribution on different cell types. Relationships, if any, will be established between occupied insulin receptor and specific cell surface structures and components. (2) to analyze the post-occupancy mobility of insulin receptors on cells that demonstrate occupancy-induced changes in receptor organization or distribution. Various chemical reagents will be used in attempts to block receptor microaggregation in order to determine the mechanisms involved and relationships to insulin action. (3) to determine the routes and subcellular structures involved in the internalization, degradation or processing, and recycling of insulin and insulin receptors. In addition to using monomeric ferritin-insulin for morphological studies, immunocytochemistry using both anti-insulin receptor antibody and biotinyl-labeled insulin binding to insulin receptors followed by antibody or avidin conjugated colloidal gold particles and/or peroxidase will be used to demonstrate the insulin receptors in intact as well as permeabilized cells. This laboratory is in a unique position to accomplish this proposal because of our proven capability to perform both quantitative morphological and correlative biochemical studies. These studies will contribute to our understanding of insulin action and of possible sites for post-binding defects in insulin resistant states.