Internal Dynamics of the Postsynaptic Density

突触后密度的内部动力学

基本信息

批准号：
10517494
负责人：
Thomas A Blanpied
金额：
$ 68.12万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-04-05 至 2024-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10517494
关键词：
AMPA Receptors Actins Action Potentials Acute Address Adhesions Alzheimer&apos s Disease Architecture Behavior Binding Biological Brain Cell Adhesion Molecules Cells Complex Cytoskeleton Data Disease Electroporation Elements Event Evoked Potentials Extracellular Domain Face Funding Genetic Glutamate Receptor Glutamates Goals Grant Human Individual Knock-out Light Link Maintenance Maps Mediating Memory Mental Depression Mental disorders Methodology Methods Modeling Molecular Movement Nanostructures Neurons Neurosciences Optics Outcome Pattern Peptide Hydrolases Performance Physiological Physiology Play Polymers Positioning Attribute Probability Process Proteins Publishing Reagent Resolution Role Sampling Scaffolding Protein Schizophrenia Signal Transduction Site Slice Structure Synapses Synaptic Cleft Synaptic Potentials Synaptic Receptors Synaptic Transmission Synaptic plasticity Technology Testing Time Vesicle Visualization Work addiction autism spectrum disorder biochemical tools brain tissue density depolymerization experience experimental study extracellular genetic regulatory protein imaging modality in utero in vivo insight molecular imaging nano nanocluster nanometer resolution nanopattern nanoscale neuroligin 1 neurotransmitter release pharmacologic polymerization postsynaptic presynaptic protein complex receptor recruit single molecule spatiotemporal superresolution imaging synaptic function tool

项目摘要

Mechanisms that create, maintain, and modulate synapses are essential building blocks of human behavior. Disruptions to these mechanisms are inextricably linked to aberrant behavior and diseases ranging from depression and schizophrenia to addiction and Alzheimer’s Disease. Thus, the long-term goal of this grant is to pursue a deep understanding of the molecular organization underlying synaptic transmission and plasticity. Our previous work took advantage of the extremely high-resolution enabled by single-molecule imaging methods and determined that at glutamatergic synapses, key proteins in the active zone and the postsynaptic density are enriched in subsynaptic nanodomains (<100 nm). Most surprisingly, nanodomains of the critical fusion-regulatory proteins RIM and Munc13 in the presynaptic active zone align with high precision across the synaptic cleft from nanodomains enriched in postsynaptic glutamate receptors. Using single-vesicle fusion mapping, we determined that the local density of RIM within active zone subregions predicts the probability of action potential-evoked vesicle fusion. This striking architectural arrangement has important implications for how synapses function. This nano-alignment between release sites and receptors can modulate synaptic transmission and potentially influence intracellular signaling. Preliminary data here and published work from others establishes that transsynaptic nanoalignment is an important element of synaptic architecture, widely present in diverse synapse types. Further, our data provide firm evidence that subsynaptic nanostructure and nanoalignment are dynamically modulated during synaptic plasticity and actively maintained by ongoing molecular interactions. These observations strongly motivate understanding the mechanisms involved in creating and maintaining transsynaptic alignment. Therefore, we will test a set of related but independent hypotheses about the origin and maintenance of transsynaptic nanoalignment. We will test 1) whether two key neurexin partners, neuroligin and LRRTM, cooperate to provide the structural basis of transsynaptic alignment, 2) whether glutamate receptors themselves are necessary or sufficient to influence the nanoscale protein organization of the active zone 3), whether the active zone RIM complex conveys instructive information to establish postsynaptic nanopatterning, and 4) how the actin cytoskeleton exerts ongoing control over synapse nanoscale architecture. To answer these questions, we have worked to establish and apply several new broadly useful technologies. We utilize a new super-resolution imaging methodology to visualize cellular substructure at nanometer resolution in vivo, apply multiplexed single-molecule imaging to map numerous proteins in the same sample, and develop new optical and biochemical tools to acutely control the actin cytoskeleton, adhesion complexes, and receptor distribution with high spatiotemporal resolution and in brain slices. The outcomes of these experiments will answer core questions about the genesis of an important new aspect of synaptic architecture and test the physiological role of synaptic nanoalignment in brain circuits.

创建，维护和调节突触的机制是人类行为的必要基础。这些机制的破坏与异常行为和疾病范围有密密相关抑郁和精神分裂症对成瘾和阿尔茨海默氏病。那，这笔赠款的长期目标是深入了解突触传播和可塑性的分子组织。我们以前的工作利用了通过单分子成像启用的极高分辨率方法并确定在谷氨酸能突触，活动区中的关键蛋白和突触后的关键蛋白密度富含突触次突触纳米域（<100 nm）。关键的最令人惊讶的纳米域融合调节蛋白RIM和MUNC13在突触前的活性区中与高精度在整个整个富含突触后谷氨酸受体的纳米域的突触裂缝。使用单维融合映射，我们确定有效区域子区域内边缘的局部密度预测了动作潜在的诱发囊泡融合。这种罢工建筑安排对突触如何功能。释放站点和接收器之间的这种纳米对齐可以调节突触传播并潜在影响细胞内信号传导。这里的初步数据，并发布了其他人则确定，经触发纳米纳米对是突触结构的重要组成部分，广泛存在于潜水员突触类型中。此外，我们的数据提供了稳健的纳米结构和在突触可塑性期间，纳米定位会动态调节，并通过持续的分子相互作用。这些观察结果强烈激发了涉及的机制创建和维护透射性一致性。因此，我们将测试一组相关但独立的关于透射性纳米定位的起源和维护的假设。我们将测试1）是否两个键神经氧蛋白伴侣，神经素和LRRTM，合作提供了透射性比对的结构基础， 2）谷氨酸受体本身是否需要或足以影响纳米级蛋白活动区3）的组织，活动区边缘复合物是否传达有指导性的信息建立突触后纳米图案，以及4）肌动蛋白细胞骨架如何施加对突触的持续控制纳米级建筑。为了回答这些问题，我们一直在努力建立并应用一些新的广泛有用的技术。我们利用一种新的超分辨率成像方法可视化细胞在体内纳米分辨率下的子结构，应用多路复用的单分子成像来绘制众多同一样品中的蛋白质，并开发新的光学和生化工具来急性控制肌动蛋白具有高时空分辨率的细胞骨架，粘合剂复合物和受体分布切片。这些实验的结果将回答有关重要新的起源的核心问题突触结构的方面并测试突触纳米定位在脑电路中的身体作用。