SYNAPTIC ARCHITECTURE, DYNAMICS AND MECHANISM

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 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，以及这些结构在神经疾病中是如何受到干扰的。我们特别感兴趣的是信号分子和酶的结构安排，以及这些分子组装的形成和在突触传递和调制过程中经历变化的方式。我们的方法是使用X射线和电子结晶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三维重建和完整化学突触的断层分析获得的EM图像相结合。我们的长期目标是为化学突触构建一个动态的分子和结构图，这将有助于理解突触的形成、传递和可塑性。利用电子断层扫描的方法，我们已经开始研究培养神经元中化学突触的构筑。我们的第一个目标是建立存在于突触的共同结构元素，并使用特定的抗体标记或基因标记来识别涉及的分子。随后，我们将执行场电位刺激和低温捕获相结合的方法来研究涉及突触传递的动态过程。最终，我们计划研究长期的、依赖刺激的突触变化，以期深入了解突触可塑性背后的建筑元素。 资金来源

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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