SYNAPTIC ARCHITECTURE, DYNAMICS AND MECHANISM

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 我们的研究重点是促进化学突触神经元之间通信的分子和超分子结构，以及这些结构如何在神经系统疾病中受到干扰。我们对信号分子和酶的结构安排特别感兴趣，并描述了这些分子组装体形成的方式，以及在突触传递和调制过程中发生的变化。我们的方法是使用X射线和电子晶体学方法研究单个蛋白质，并将这些信息与通过超分子组装的3-D重建和完整化学突触的断层扫描分析获得的EM图像联合收割机相结合。我们的长期目标是构建一个化学突触的动态分子和结构图，这将有助于理解突触的形成，传递和可塑性。利用电子断层摄影的方法，我们已经开始研究的化学突触在培养的神经元的架构。我们的第一个目标是建立共同的建筑元素目前在突触和识别分子参与使用特异性抗体标记或遗传标记。随后，我们将进行场电位刺激结合低温捕获来研究突触传递中涉及的动态过程。最终，我们计划研究长期的，刺激依赖的，突触的变化，希望深入了解突触可塑性的基础架构元素。 资金

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Our research is focused on molecular and supramolecular structures that facilitate communication between neurons at the chemical synapse and how such structures are perturbed in neurological disease. We are particularly interested in the architectural arrangement of signaling molecules and enzymes, and characterizing the ways in which such molecular assemblies are formed and undergo changes during synaptic transmission and modulation. Our approach is to investigate individual proteins using x-ray and electron crystallographic methods and to combine this information with EM images obtained via 3-D reconstruction of supramolecular assemblies and tomographic analysis of the intact chemical synapse. Our long-term goal is to construct a dynamic molecular and architectural map for the chemical synapse that will help to understand synaptic formation, transmission and plasticity. Using electron tomographic methods we have begun to study the architecture of the chemical synapse in cultured neurons. Our first goal is to establish the common architectural elements present at the synapse and to identify the molecules involved using specific antibody labeling or genetic tagging. Subsequently, we will perform field potential stimulations coupled with cryogenic trapping to investigate the dynamic processes involved in synaptic transmission. Ultimately we plan to study long-term, stimulation dependent, synaptic changes in the hopes of gaining insight into the architectural elements underlying synaptic plasticity. FUNDING