INSULIN INDUCTION OF GLUCOSE TRANSPORTER TRANSLOCATION

申请人提出一个研究和学习计划，为他做好准备， 从事内分泌疾病的研究第一阶段研究 将在Harvey F博士的实验室进行。洛迪什在怀特黑德 生物医学研究所和马萨诸塞州 技术.申请人将研究胰岛素导致 脂肪和肌肉细胞吸收和利用葡萄糖。这一主题具有广泛的 对II型糖尿病的病理生理学的影响， 至于神经内分泌激素的作用， 水重吸收和细胞代谢的控制。 特别地，申请人感兴趣的是通过以下方式阐明该机制： 胰岛素与其受体的结合诱导囊泡融合 含有GLUT 4葡萄糖转运蛋白与质膜。因为 胰岛素敏感性细胞中的囊泡运输 组织，神经突触和酵母，这个问题在几个方面是可以解决的。 程度.申请人将使用培养的3 T3-L1脂肪细胞， 第一阶段，并计划表征已知的蛋白质同源物， 对于其他细胞类型中囊泡转运重要，以及新的 在脂肪细胞分化过程中被强烈诱导的基因。等 基因包括小GTP结合蛋白Rab 3A、Rab 3D、Rab 4和Rab 5， 突触泡蛋白和小突触泡蛋白（VAMP）的同源物， syntaxin和caveolin。申请方将建立一种测定方法， 将GLUT 4易位至质膜，并将构建载体 以驱动脂肪细胞特异性基因表达， 转染和永久细胞系。这些工具可以让他 过表达各种突变基因或反义RNA，并评估 这些干预措施对胰岛素刺激的葡萄糖摄取的影响。的 申请人还将能够分析胰岛素刺激的 几种蛋白质，包括adipsin和一种可能的C1 q同源物， 补体亚单位蛋白在较长的项目中，申请人 他建议使用他开发的工具来寻找新的基因 对胰岛素触发的葡萄糖摄取很重要。

The applicant proposes a program of research and study to prepare him for a career in the investigation of endocrine diseases. The phase 1 research will be conducted in the laboratory of Dr. Harvey F. Lodish at Whitehead Institute for Biomedical Research and Massachusetts Institute of Technology. The applicant will study the mechanism by which insulin causes fat and muscle cells to take up and utilize glucose. This topic has broad implications for the pathophysiology of Type II Diabetes Mellitus, as well as for neuroendocrine hormone action, vasopressin regulation of renal water reabsorption, and control of cellular metabolism. In particular, the applicant is interested in elucidating the mechanism by which binding of insulin to its receptor induces fusion of vesicles containing the GLUT4 glucose transporter with the plasma membrane. Because of the similarities between vesicular trafficking in insulin-responsive tissues, nerve synapses, and yeast, the problem is approachable on several levels. The applicant will work with cultured 3T3-L1 adipocytes during phase I, and plans to characterize homologues of proteins known to be important for vesicle transport in other cell types, as well as novel genes that are strongly induced during adipocyte differentiation. Such genes include the small GTP-binding proteins Rab3A, Rab3D, Rab4 and Rab5, homologues of synaptophysin and synaptobrevin (VAMP), an isoform of syntaxin, and caveolin. The applicant will establish an assay for translocation of GLUT4 to the plasma membrane, and will construct a vector to drive adipocyte-specific gene expression in both transient transfections and permanent cell lines. These tools will allow him to overexpress various mutant genes or antisense RNAs, and to assess the effect of these interventions on insulin-stimulated glucose uptake. The applicant will also be able to analyze insulin-stimulated secretion of several proteins, including adipsin and a likely homologue of a C1q complement subunit protein. In a longer range project, the applicant proposes to use the tools he will have developed to search for novel genes important for insulin-triggered glucose uptake.