Molecular mechanisms of GLURl trafficking and insertion

GLUR1运输和插入的分子机制

• 批准号: 6445835
• 负责人: GAVIN R RUMBAUGH
• 金额: $ 3.83万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6445835
• 关键词: AMPA receptors; electrophysiology; long term potentiation; nerve /myelin protein; neural transmission; neurophysiology; protein 4.1; protein protein interaction; protein structure function; protein transport; receptor expression; tissue /cell culture; tumor suppressor proteins

DESCRIPTION (provided by applicant): The main focus of this research proposal is to elucidate the molecular machinery involved in trafficking of AMPA receptors during both developmental and activity dependent paradigms. This study will further the understanding of how pre synaptic signals can lead to acute as well was as long-term changes in synaptic efficacy. This proposal has the advantage of utilizing a wide range of techniques to address the important questions posed. Using a combination of electrophysiological, biochemical, molecular, and microscopy methods, interactions of the GluR1 AMPA receptor subunit with intracellular synaptic proteins can be implicated as key players in synaptic function. A primary neocortical culture system will be used as a model for a functional neuronal network. Overexpression of the wildtype or mutant fluorescent-tagged AMPA receptor interacting proteins SAP97/hDLG and Protein 4.1 by the Sindbis viral expression system will enable the study of these proteins with regard to their effect on synaptic transmission. The effect of chronic (>24 HR) overexpression of SAP97 and Protein 4.1 will be assessed by measuring AMPA mediated mEPSCs. Also, surface expression of GluR1 will also be measure by biochemical methods after overexpression of these proteins. Finally, using a novel assay for long-term potentiation in cultured neurons overexpressing either normal or mutant forms of SAP97 and Protein 4.1 will be employed to understand the role of these proteins in synaptic plasticity.

描述（由申请人提供）： 本研究计划的主要重点是阐明分子 发育过程中参与 AMPA 受体运输的机制 和活动依赖范例。这项研究将进一步加深对 突触前信号如何导致急性以及长期的变化 突触功效。 This proposal has the advantage of utilizing a wide range of 解决所提出的重要问题的技术。使用组合 电生理学、生化、分子和显微镜方法， GluR1 AMPA 受体亚基与细胞内突触的相互作用 蛋白质可能是突触功能的关键参与者。初级 新皮质培养系统将用作功能性神经元的模型 网络。野生型或突变型荧光标记 AMPA 的过度表达 Sindbis 病毒的受体相互作用蛋白 SAP97/hDLG 和蛋白 4.1 表达系统将使这些蛋白质的研究成为可能 对突触传递的影响。慢性（>24 HR）过度表达的影响 SAP97 和蛋白质 4.1 的含量将通过测量 AMPA 介导的 mEPSC 进行评估。 此外，GluR1的表面表达也将通过生化方法进行测量 这些蛋白质过度表达后。最后，使用一种新颖的检测方法 培养的神经元过度表达正常或的长期增强 SAP97 和蛋白质 4.1 的突变形式将用于了解其作用 这些蛋白质在突触可塑性中的作用。