INSULIN INDUCTION OF GLUCOSE TRANSPORTER TRANSLOCATION

胰岛素诱导葡萄糖转运蛋白易位

• 批准号: 2134306
• 负责人: JONATHAN BOGAN
• 金额: $ 6.8万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-08-10 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2134306
• 关键词: 3T3 cells; adipocytes; biological signal transduction; cell membrane; clone cells; complement; confocal scanning microscopy; fluorescence microscopy; gene expression; glucose transporter; hormone regulation /control mechanism; insulin; insulin receptor; laboratory mouse; mutant; receptor binding; secretion; transfection; vesicle /vacuole

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.

申请人提出了一项研究和研究计划，以准备他 研究内分泌疾病的职业。第一阶段研究 将在Whitehead的Harvey F. Lodish博士的实验室进行 生物医学研究所和马萨诸塞州研究所 技术。申请人将研究胰岛素引起的机制 脂肪和肌肉细胞占用并利用葡萄糖。这个话题广泛 对II型糖尿病的病理生理的影响以及 至于神经内分泌激素作用，血管加压素调节肾脏 水的吸收和细胞代谢的控制。 特别是，申请人有兴趣通过 胰岛素与其受体结合诱导囊泡的融合 含有质膜的GLUT4葡萄糖转运蛋白。因为 胰岛素响应性的囊泡贩运之间的相似之处 组织，神经突触和酵母，几个问题都可以接近 水平。申请人将与经过培养的3T3-L1脂肪细胞一起工作 第一阶段，并计划表征已知的蛋白质的同源物 对于其他细胞类型的囊泡运输以及新颖的 在脂肪细胞分化过程中强烈诱导的基因。这样的 基因包括小的GTP结合蛋白Rab3a，rab3d，rab4和rab5， 突触蛋白和突触的同源物（VAMP），这是一种同工型 语法素和小窝蛋白。申请人将为 GLUT4转移到质膜，将构建矢量 在两个瞬态中驱动脂肪细胞特异性基因表达 转染和永久性细胞系。这些工具将使他能够 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。这 申请人还将能够分析胰岛素刺激的分泌 几种蛋白质，包括脂肪素和C1Q的可能同源物 补体亚基蛋白。在一个长期项目中，申请人 建议使用他将开发的工具来搜索新型基因 对于胰岛素触发的葡萄糖摄取至关重要。