Probing the Structure of the Synapse Using Superresolution Light Microscopy

使用超分辨率光学显微镜探测突触的结构

基本信息

批准号：
7340350
负责人：
GINA G TURRIGIANO
金额：
$ 77.58万
依托单位：
BRANDEIS UNIVERSITY
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：
2007-09-30 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7340350
关键词：
3-Dimensional AMPA Receptors Accounting Address Alzheimer&apos s Disease Area Attention Autistic Disorder Binding Biochemical Biochemical Genetics Biological Assay Biology Biophysics Brain COS-7 Cell Cartoons Cells Chemicals Chromosome Pairing Class Collaborations Complex Computer Simulation Computing Methodologies Creativeness Culture Techniques Data Depth Development Dimensions Electron Microscopy Electrons Ensure Environment Equilibrium Ethane Event Excitatory Synapse Face Faculty Fellowship Fire - disasters Fluorescence Fluorescence Microscopy Freezing Funding Future Gases Glutamate Receptor Goals Green Fluorescent Proteins Health Heavy Metals Homeostasis Image Imagery Immersion Investigative Technique Immobilization In Situ Individual Influentials Information Storage Invertebrates Knock-in Mouse Label Learning Light Light Microscope Liquid substance Localized Location Long-Term Potentiation Maps Measurement Measures Membrane Proteins Memory Methane Methodology Methods Microscope Microscopy Microtomy Modeling Molecular Molecular Genetics Monitor Nature Neuronal Plasticity Neurons Neurosciences Neurotransmitter Receptor Nitrogen Noise Numbers Optics Pattern Photons Physiologic pulse Physiology Pliability Poisson Distribution Positioning Attribute Postdoctoral Fellow Postsynaptic Membrane Preparation Procedures Process Property Protein Kinase Protein Overexpression Proteins Publishing Pulse taking Purpose Range Refractive Indices Regulation Relative (related person)Research Resolution Risk Role Sampling Science Shoulder Side Signal Transduction Signaling Molecule Site Slice Slide Source Specimen Staging Standards of Weights and Measures Structural Biologist Structure Surface Synapses Synaptic Receptors Synaptic plasticity System Techniques Technology Temperature Testing Thick Thinking Time Variant Visual Cortex Water Weight Width Work abstracting addiction base calmodulin-dependent protein kinase II central pattern generator cognitive function cold temperature density experience fluorescence imaging fluorophore hippocampal pyramidal neuron in vivo instrument instrumentation interest intracellular protein transport lens light microscopy member molecular array neocortical nervous system disorder new technology novel novel strategies postsynaptic postsynaptic density protein presynaptic density protein 95 programs protein localization location protein protein interaction receptor receptor density response sample fixation size skills sleep epilepsy small molecule spatial relationship statistics structural biology synaptic function synergism technology development tool trafficking visual deprivation yeast two hybrid system

项目摘要

Memory and other cognitive functions reside in part in the pattern and strength of synaptic connections between neurons. Understanding the molecular determinants of synaptic strength has been a longstanding goal of neuroscience, and advances in this field stand to influence our understanding of virtually every neurological disorder from Autism to Alzheimer?s disease. Over the past decade biochemical and conventional molecular and genetic approaches have begun to piece together how interactions between neurotransmitter receptors and other synaptic proteins regulate and control synaptic strength and plasticity, but a major limitation is that there is little or no structural information about how proteins are arranged into signaling complexes at the synapse. Many signaling molecules can only interact with immediately adjacent proteins, and this localization may itself be regulated by experience. Understanding how functional signaling complexes are generated and how they in turn regulate synaptic strength thus requires that we probe the spatial arrangements of proteins within the postsynaptic density (PSD). Conventional approaches do not have sufficient resolution to allow the position of synaptic proteins to be mapped within these tiny (< 1 im) synaptic structures. Here I propose to develop tools to map the spatial arrangements of individual synaptic proteins (such as glutamate receptors) within the PSD, and to determine how these spatial arrangements are influenced by synaptic plasticity, using super resolution light microscopy. By mapping the relative positions of many different proteins within the postsynaptic membrane and PSD we will be able to generate a 3 dimensional model of the protein lattices that comprise the postsynaptic side of the synapse. This method has the promise to put a vast array of biochemical and molecular data on protein-protein interactions into a structural context that is essential for its interpretation, and will add a powerful new tool to the analysis of synaptic function.

记忆和其他认知功能部分取决于神经元之间突触连接的模式和强度。了解突触强度的分子决定因素一直是神经科学的长期目标，该领域的进步将影响我们对从自闭症到阿尔茨海默氏病的几乎所有神经系统疾病的理解。在过去的十年中，生物化学和传统的分子和遗传学方法已经开始拼凑神经递质受体和其他突触蛋白之间的相互作用如何调节和控制突触强度和可塑性，但主要的限制是关于蛋白质如何作用的结构信息很少或没有。在突触处排列成信号复合体。许多信号分子只能与直接相邻的蛋白质相互作用，并且这种定位本身可能受到经验的调节。因此，了解功能性信号复合物是如何产生的以及它们如何调节突触强度，需要我们探测突触后密度（PSD）内蛋白质的空间排列。传统方法没有足够的分辨率来在这些微小（< 1 im）突触结构内绘制突触蛋白的位置。在这里，我建议开发工具来绘制 PSD 内单个突触蛋白（例如谷氨酸受体）的空间排列，并使用超分辨率光学显微镜确定这些空间排列如何受到突触可塑性的影响。通过绘制突触后膜和 PSD 内许多不同蛋白质的相对位置，我们将能够生成构成突触后侧的蛋白质晶格的 3 维模型。这种方法有望将大量有关蛋白质-蛋白质相互作用的生化和分子数据放入对其解释至关重要的结构背景中，并将为突触功能分析添加一个强大的新工具。