Mechanisms of neurotransmitter release and its regulation

神经递质释放机制及其调控

• 批准号: 10057389
• 负责人: Jose Rizorey
• 金额: $ 88.55万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10057389
• 关键词: Acute; Alzheimer' s Disease; Biological Assay; Blood Pressure; Brain; Cellular biology; Communication; Complement; Complex; Cryo-electron tomography; Cryoelectron Microscopy; Defect; Development; Disease; Docking; Electrophysiology (science); Goals; Heart Rate; Insulin; Intracellular Membranes; Knowledge; Membrane; Membrane Fusion; Molecular; NMR Spectroscopy; Neurons; Parkinson Disease; Pharmaceutical Preparations; Physiological; Process; Proteins; Regulation; Research; S-nitro-N-acetylpenicillamine; SNAP receptor; Schizophrenia; Structural Protein; Structure; Synaptic Transmission; Synaptic Vesicles; Testing; X-Ray Crystallography; biophysical techniques; experimental study; information processing; insight; interdisciplinary approach; nervous system disorder; neurotransmitter release; novel; novel strategies; presynaptic; protein complex; protein reconstitution; reconstitution; success; synaptotagmin; therapy development; three dimensional structure

Neurotransmitter release is acutely triggered by Ca2+ and is regulated during presynaptic plasticity processes that underlie some forms of information processing in the brain. Characterization of the mechanisms of release and its regulation is thus critical to understand brain function and will facilitate the development of therapies for multiple neurological disorders (e.g. schizophrenia, Alzheimer's and Parkinson's) and for diseases involving defects in regulated secretion, which controls many important physiological functions (e.g. heart rate, blood pressure and insulin release). This research is also relevant to cell biology in general because of its importance to understand intracellular membrane fusion. The machinery that controls release contains a core formed by SNARE proteins, Munc18, Munc13, NSF and SNAP, and specialized proteins that regulate release including Munc13 itself, synaptotagmin, complexin, RIM, Rab3 and CAPS among others. The research proposed in this application involves an interdisciplinary approach integrating structural studies of these proteins, reconstitution assays and electrophysiological analyses of neurotransmitter release in neurons performed by collaborators. This research will build on the previous success of this approach, which has yielded many of the three-dimensional structures of the proteins that govern neurotransmitter release, has revealed crucial mechanistic concepts in the field and has allowed reconstitution of basic steps of Ca2+-evoked synaptic vesicle fusion with eight central components of the release machinery. However, despite these advances, the mechanism of release is still unclear and fundamental questions remain unanswered. The ultimate goals of this proposal are to develop a detailed picture of the mechanism of release that integrates the functions of all these proteins and to provide novel insights into how release is regulated in diverse presynaptic plasticity processes. For this purpose, structural studies of complexes formed by these proteins using a combination of biophysical techniques, including cryo-electron microscopy, cryo-electron tomography, NMR spectroscopy and X-ray crystallography, will be performed. A key aspect of these studies, which is essential to make major, definitive advances in this field, will be to analyze protein complexes between two membranes, as the membranes form intrinsic part of the fusion apparatus and thus are expected to have a strong influence on how the protein components are arranged to induce membrane fusion. The structural studies will be complemented with reconstitution experiments to understand how the different proteins control docking, priming and fusion, as well as with physiological analyses performed by collaborators that will test the relevance of the structural and reconstitution studies. This research is expected to establish fundamental principles on neuronal communication that are vital for brain function.

神经递质释放是由Ca2+急剧触发的，并在突触前可塑性过程中受到调节 这是大脑中某些形式的信息处理。释放机制的表征 因此，它的调节对于了解大脑功能并促进疗法的发展至关重要 用于多种神经系统疾病（例如精神分裂症，阿尔茨海默氏症和帕金森氏症），以及涉及的疾病 受调节分泌的缺陷，它控制着许多重要的生理功能（例如心率，血液 压力和胰岛素释放）。这项研究也与细胞生物学有关，因为它的重要性 了解细胞内膜融合。控制释放的机械包含由 SNARE蛋白，Munc18，Munc13，NSF和SNAP，以及调节释放的专门蛋白质 munc13本身，突触量，复合蛋白，边缘，rab3和帽子等。这项研究提出了 应用涉及一种跨学科方法，整合了这些蛋白质的结构研究， 在由神经递质释放神经元释放神经元中的重构测定和电生理分析 合作者。这项研究将以这种方法的先前成功为基础，该方法产生了许多 控制神经递质释放的蛋白质的三维结构已显示至关重要 该领域的机械概念，并允许重组Ca2+诱发的突触囊泡的基本步骤 融合与发行机械的八个中央组件。但是，尽管有这些进步， 释放机制仍然不清楚，基本问题仍未得到解决。最终目标 提案是为了整合所有这些功能的释放机制的详细图片 蛋白质并提供新的见解，以了解如何在多种突触前可塑性过程中调节释放。 为此，使用生物物理的组合，这些蛋白质形成的复合物的结构研究 技术，包括冷冻电子显微镜，冷冻电子断层扫描，NMR光谱和X射线 晶体学将进行。这些研究的一个关键方面，这对于制造主要的，确定的至关重要 该领域的进步将是分析两个膜之间的蛋白质复合物，为膜形式 融合设备的固有部分，因此有望对蛋白质的蛋白质产生强大的影响 组件安排以诱导膜融合。结构研究将与 重组实验以了解不同蛋白质如何控制对接，启动和融合，作为 以及合作者进行的生理分析，这些分析将测试结构和 重建研究。预计这项研究将建立有关神经元的基本原则 对大脑功能至关重要的沟通。