Defining the Role of Lipid Rafts in Neurotransmission

定义脂筏在神经传递中的作用

• 批准号: 7107251
• 负责人: Alice D Lam
• 金额: $ 3.17万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7107251
• 关键词: SDS polyacrylamide gel electrophoresis; calcium channel; cell line; chromaffin cells; cyclodextrins; dielectric property; exocytosis; flash photolysis; membrane fusion; membrane structure; neural transmission; predoctoral investigator; protein localization; protein protein interaction; syntaxin; voltage gated channel

DESCRIPTION (provided by applicant): SNARE proteins form the minimal machinery required for regulated exocytosis, a process that underlies all information transfer within the nervous system. Lipid rafts, which have been implicated in a number of diseases including Alzheimer's and Parkinson's, are dynamic membrane microdomains that serve as platforms to regulate membrane function. While SNARE proteins have been found to localize to lipid rafts, and while disruption of lipid rafts results in the inhibition of regulated exocytosis, the exact role that lipid rafts exert in mediating exocytosis has not yet been determined. This proposal seeks to: 1) identify the specific stages of regulated exocytosis (e.g., core complex formation, priming, and fusion) in which lipid rafts play an important role; 2) determine the function of lipid rafts in modulating fusion pore dynamics; and 3) determine the regulatory elements involved in targeting of the neuronal SNARE protein Syntaxin 1A to lipid raft domains. The experimental approach will utilize a combination of carbon fiber amperometry and membrane capacitance measurements in both intact and cholesterol-depleted chromaffin cells, as well as biochemical lipid raft isolations in HEK cells heterologously expressing various wild-type and mutant SNARE proteins.

描述（由申请人提供）：SNARE 蛋白形成调节胞吐作用所需的最小机制，胞吐作用是神经系统内所有信息传递的基础过程。脂筏与包括阿尔茨海默氏症和帕金森氏症在内的许多疾病有关，是动态的膜微域，可作为调节膜功能的平台。虽然已发现 SNARE 蛋白定位于脂筏，并且脂筏的破坏会导致受调节的胞吐作用受到抑制，但脂筏在介导胞吐作用中发挥的确切作用尚未确定。该提案旨在：1）确定脂筏在受调节的胞吐作用（例如核心复合物形成、启动和融合）中发挥重要作用的具体阶段； 2）确定脂筏在调节融合孔动力学中的功能； 3) 确定神经元 SNARE 蛋白 Syntaxin 1A 靶向脂筏结构域所涉及的调控元件。该实验方法将在完整和胆固醇耗尽的嗜铬细胞中结合使用碳纤维电流分析法和膜电容测量，以及在异源表达各种野生型和突变型 SNARE 蛋白的 HEK 细胞中进行生化脂筏分离。