Neurotransmitter release machinery: structure and function

神经递质释放机制：结构和功能

• 批准号: 7807057
• 负责人: Jose Rizorey
• 金额: $ 40.56万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-12-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7807057
• 关键词: 1,2-diacylglycerol; Address; Affect; Alzheimer' s Disease; Binding; Biochemical; Biological Assay; Brain; C-terminal; C2 Domain; Cell membrane; Communication; Complex; Coupling; Cryoelectron Microscopy; DAG/PE-Binding Domain; Data; Diglycerides; Disease; Drug Addiction; Exocytosis; Goals; Grant; Homologous Gene; Intracellular Membranes; Laboratories; Lead; Left; Lipids; Liposomes; Mediating; Membrane; Membrane Fusion; Methods; Modeling; Molecular Conformation; Munc18-1 protein; Mus; Mutagenesis; Mutation; N-terminal; NMR Spectroscopy; Neurons; Parkinson Disease; Phosphorylation; Phosphorylation Site; Play; Process; Protein Binding; Proteins; Publishing; Regulation; Research; Resolution; Roentgen Rays; Role; SNAP receptor; Structure; Synaptic Transmission; Synaptic Vesicles; Techniques; Testing; Vesicle; X-Ray Crystallography; base; brain disorder therapy; complement C2a; complement C2b; complexin I; density; design; in vivo; information processing; insight; interdisciplinary approach; neurotransmitter release; novel; presynaptic; protein complex; protein function; reconstitution; research study; success; synaptosomal-associated protein 25; synaptotagmin I; syntaxin; syntaxin 1; syntaxin binding protein 1; therapy design; vesicle-associated membrane protein

DESCRIPTION (provided by applicant): Synaptic transmission in the brain is initiated by neurotransmitter release, which is mediated by synaptic vesicle exocytosis. Insights into the mechanisms of neurotransmitter release and its regulation are essential for understanding how the brain processes information, and how synaptic transmission is affected in diseases such as Parkinson's disease, Alzheimer's disease, and drug addiction. Release is governed by a complex protein machinery that includes as central components proteins with homologues in most types of intracellular membrane fusion such as the SNARE proteins and munc18-1. In addition, several proteins such as munc13-1 and complexins play critical roles that are specialized for the tight spatial and temporal regulatory requirements of neurotransmitter release. While the research performed under this grant and studies from other laboratories have yielded key insights into how these proteins function, the mechanism of release is still unclear and fundamental questions remain unanswered. The research proposed in this application is designed to build on this success and on crucial findings made in preliminary experiments that suggest novel interactions between these key components and new hypotheses on how they function. These include an interaction between munc18-1 and the SNAREs that suggest a novel model for the core of the fusion machinery, and a munc18-1/munc13-1 interaction that suggest a novel mechanism for the priming step that leaves synaptic vesicles ready for neurotransmitter release. This research involves an interdisciplinary approach integrating structural studies at atomic resolution, biochemical experiments, reconstitution assays and electrophysiological analyses of neurotransmitter release in neurons. Four specific aims are proposed that focus on: 1. Munc 18-1/SNARE interactions; 2. Mechanisms of munc13-1 action and its regulation; 3. Complexin/SNARE interactions; and 4. Reconstitution of basic steps of neurotransmitter release. The results of this research will not only be important to understand the mechanisms of neurotransmitter release and membrane fusion in general, but will also have an impact in the design of therapies for brain disorders that involve changes in synaptic transmission.

描述（由申请人提供）：大脑中的突触传播是由神经递质释放引发的，神经递质释放是由突触囊泡胞吐介导的。对神经递质释放机制及其调控的见解对于了解大脑如何处理信息以及在诸如帕金森氏病，阿尔茨海默氏病和吸毒等疾病中如何影响突触传播至关重要。释放受复杂的蛋白质机械的控制，该机制包括大多数类型的细胞内膜融合中的中央成分蛋白，例如SNARE蛋白和Munc18-1。此外，几种蛋白质（例如Munc13-1）和复合素扮演着关键作用，这些作用专门用于神经递质释放的紧密空间和时间调节要求。尽管该赠款进行的研究和其他实验室的研究对这些蛋白质的功能产生了关键的见解，但释放的机制仍然不清楚，基本问题仍然没有得到解决。本应用程序中提出的研究旨在建立在这一成功的基础上，并基于初步实验中提出的关键发现，这些发现暗示了这些关键组成部分之间的新型相互作用和关于它们如何运作的新假设。其中包括Munc18-1和网罗之间的相互作用，该杂志暗示了融合机械核心的新型模型，以及Munc18-1/Munc13-1的相互作用，这暗示了启动步骤的新型机制，该机制留下了为神经递质释放准备的突触囊泡。这项研究涉及一种跨学科的方法，该方法在原子分辨率，生化实验，重建测定和神经递质释放的神经递质释放的电生理分析中整合结构研究。提出了四个具体目标，重点是：1。MUNC18-1/SNARE ITSOTIONS； 2。Munc13-1作用及其调节的机制； 3。和4。神经递质释放的基本步骤的重构。这项研究的结果不仅要了解神经递质释放和膜融合的机制，而且还将对涉及突触传播变化的脑部疾病的设计产生影响。