Organization and Function of the Periactive Zone

周围活动区的组织和功能

基本信息

批准号：
10600083
负责人：
Avital Adah Rodal
金额：
$ 37.99万
依托单位：
BRANDEIS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-15 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10600083
关键词：
Actins Affect Architecture Binding Biochemical Biological Biological Assay Cell Adhesion Cell Adhesion Molecules Cell physiology Cells Clathrin Communication Complex Cytoskeleton Data Disease Drosophila genus Endocytosis Etiology Event Geometry Goals Homologous Gene Image Individual Learning Link Maps Mediating Membrane Methods Microfilaments Modeling Monitor Morphogenesis Neuroglia Neuromuscular Junction Neurons Neurotransmitters Polymers Probability Process Property Proteins Recycling Regulation Rest Reticulum Role Site Structure Synapses Synaptic Membranes Synaptic Transmission Synaptic Vesicles System Techniques Testing Time Work experimental study genetic testing high resolution imaging human disease imaging modality in vivo mutant nervous system disorder particle polymerization presynaptic receptor response segregation spatiotemporal superresolution imaging synaptic function target SNARE proteins tool transmission process vesicular release

项目摘要

PROJECT SUMMARY The goal of this proposal is to understand the spatial and functional organization of the presynaptic periactive zone (PAZ), which is found adjacent to sites of synaptic vesicle release. The PAZ is a micron-scale structure, occupied by dozens of proteins that work together in multivalent assemblies to couple membrane remodeling to force-generating actin polymerization. Studies of PAZ proteins in many systems have suggested that these proteins act at multiple steps of the synaptic vesicle cycle as well as in other synaptic membrane functions (e.g. synaptic morphogenesis and receptor traffic). It remains unknown how the micron-scale organization and regulation of PAZ proteins direct their membrane and cytoskeleton remodeling activities to these different neuron-specific functions. We will use the Drosophila larval neuromuscular junction (NMJ), a powerful model synapse, to decipher how PAZ protein assemblies, activities, and cellular functions are linked. Using high- resolution imaging, we recently found that PAZ proteins occupy both overlapping and distinct domains within the PAZ, and that proper segregation of PAZ proteins between these domains depends on their multivalent interactions with each other. We have also recently described PAZ-dependent dynamic actin filament structures, which represent a direct readout of PAZ protein activities in these different domains. Using these tools, we will ask how synapses regulate PAZ protein activities and interactions in space and time, and how PAZ organization underlies its diverse neuron-specific functions, in response to synaptic activity and transmission. In Aim 1, we will determine how PAZ proteins are organized at resting and active synapses using complementary fixed, live, and super-resolution imaging methods, and develop new quantitative methods to describe their geometric relationships in PAZ domains. In Aim 2, we will test the hypothesis that synaptic actin patches represent clathrin-dependent synaptic vesicle recycling events, and identify the determinants of synaptic actin patch assembly and dynamics. In Aim 3, we will ask how PAZ organization and synaptic activity control diverse cell biological PAZ functions, including release site clearance and organization of cell adhesion complexes. Overall, our experiments will explain how organization of the PAZ into distinct subdomains underlies multiple functional and structural properties of synapses. PAZ proteins are implicated in multiple neurological disorders, so deciphering their in vivo functions will be critical for understanding the etiology of these human diseases.

项目概要该提案的目标是了解突触前周围活动的空间和功能组织区域（PAZ），位于突触小泡释放位点附近。 PAZ是微米级结构，被数十种蛋白质占据，这些蛋白质在多价组装中共同作用以耦合膜重塑到产生力的肌动蛋白聚合。对许多系统中 PAZ 蛋白的研究表明，这些蛋白质在突触小泡循环的多个步骤以及其他突触膜功能中发挥作用（例如突触形态发生和受体交通）。目前尚不清楚微米级的组织和 PAZ 蛋白的调节将其膜和细胞骨架重塑活动引导至这些不同的神经元特异性功能。我们将使用果蝇幼虫神经肌肉接头 (NMJ)，这是一个强大的模型突触，破译 PAZ 蛋白组装、活性和细胞功能之间的联系。使用高分辨率成像，我们最近发现 PAZ 蛋白占据了重叠和不同的域 PAZ，这些结构域之间 PAZ 蛋白的正确分离取决于它们的多价彼此之间的互动。我们最近还描述了 PAZ 依赖性动态肌动蛋白丝结构，代表这些不同域中 PAZ 蛋白活性的直接读数。使用这些工具，我们将询问突触如何调节 PAZ 蛋白活性以及空间和时间上的相互作用，以及如何 PAZ 组织是其多样化神经元特异性功能的基础，响应突触活动和传播。在目标 1 中，我们将使用以下方法确定 PAZ 蛋白在静息和活动突触中的组织方式：互补的固定、实时和超分辨率成像方法，并开发新的定量方法描述它们在 PAZ 域中的几何关系。在目标 2 中，我们将检验突触肌动蛋白的假设斑块代表网格蛋白依赖性突触小泡回收事件，并确定其决定因素突触肌动蛋白补丁组装和动力学。在目标 3 中，我们将询问 PAZ 的组织和突触活动如何控制多种细胞生物 PAZ 功能，包括释放位点清除和细胞粘附组织复合物。总的来说，我们的实验将解释如何将 PAZ 组织成不同的子域是突触多种功能和结构特性的基础。 PAZ 蛋白参与多种神经系统疾病，因此破译其体内功能对于理解神经系统疾病的病因至关重要这些人类疾病。