Mechanisms of neurotransmitter release and its regulation

神经递质释放由钙离子强烈触发，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的基础。释药机制的表征 因此，它的调节对于了解大脑功能至关重要，并将促进治疗方法的发展 适用于多种神经系统疾病(如精神分裂症、阿尔茨海默氏症和帕金森氏症)，以及涉及以下疾病的疾病 调节分泌的缺陷，它控制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，因为它很重要。 了解细胞内膜融合情况。控制释放的机械包含由以下组成的核心 SNARE蛋白、Munc18、Munc13、NSF和SNAP，以及调节释放的特殊蛋白，包括 Munc13本身、突触素、复合蛋白、RIM、Rab3和Caps等。这项研究中提出的 应用涉及整合这些蛋白质的结构研究的跨学科方法， 神经递质释放的重建分析和电生理分析 合作者。这项研究将建立在这种方法之前的成功基础上，这种方法已经产生了许多 控制神经递质释放的蛋白质的三维结构揭示了至关重要的 该领域中的机械概念，并允许重建钙离子诱导的突触小泡的基本步骤 融合了释放机械的八个中心部件。然而，尽管有这些进步， 释放机制仍不清楚，根本问题仍未得到回答。这样做的最终目标是 提议是制定一幅详细的释放机制图景，将所有这些功能整合在一起 并为如何在不同的突触前可塑性过程中调节释放提供新的见解。 为此，利用生物物理的组合对这些蛋白质形成的复合体的结构进行研究 技术，包括低温电子显微镜、低温电子断层扫描、核磁共振光谱和X射线 将进行结晶学研究。这些研究的一个关键方面，这是使主要的、明确的 这一领域的进展将是随着膜的形成，分析两层膜之间的蛋白质复合体 融合装置的固有部分，因此预计将对蛋白质如何 布置组件以诱导膜融合。结构研究将得到以下补充 重组实验，以了解不同的蛋白质如何控制对接、启动和融合，如 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。这项研究有望确立神经元的基本原理。 对大脑功能至关重要的交流。

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.