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：
7667163
负责人：
GINA G TURRIGIANO
金额：
$ 78.5万
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/7667163
关键词：
3-Dimensional AMPA Receptors Accounting Address Alzheimer&apos s Disease Area Attention Autistic Disorder Binding Biochemical Biological Assay Biology Biophysics Brain COS-7 Cell Cartoons Cells Chemicals Collaborations Complex Computer Simulation Computing Methodologies Creativeness Culture Techniques Data Development Dimensions Electron Microscopy Electrons Ensure Environment Equilibrium Ethane Event Excitatory Synapse Face Faculty Fellowship Fluorescence Fluorescence Microscopy Freezing Funding Future Gases Genetic Glutamate Receptor Goals 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 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 Optics Pattern Photons Physiologic pulse Physiology Poisson Distribution Positioning Attribute Postdoctoral Fellow Postsynaptic Membrane Preparation Procedures Process Property Protein Kinase Proteins Publishing Refractive Indices Regulation Relative (related person)Research Resolution Risk Role Sampling Science Shoulder Side Signal Transduction Signaling Molecule Site Slice Slide Source Specimen Staging 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 flexibility fluorescence imaging fluorophore hippocampal pyramidal neuron in vivo instrument instrumentation interest lens light microscopy member molecular array neocortical nervous system disorder new technology novel novel strategies overexpression postsynaptic postsynaptic density protein presynaptic density protein 95 programs protein complex protein protein interaction receptor receptor density response sample fixation 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

ABSTRACT 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 long- standing 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 ¿m) 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.

抽象的记忆和其他认知功能部分取决于突触连接的模式和强度了解神经元之间的突触强度的分子决定因素一直是一个长期的问题。神经科学的长期目标以及该领域的进步将影响我们对在过去的十年里，几乎所有的神经系统疾病，从自闭症到阿尔茨海默病。生物化学和传统的分子和遗传学方法已经开始拼凑如何神经递质受体和其他突触蛋白之间的相互作用调节和控制突触强度和塑性，但一个主要限制是很少或根本没有关于如何许多信号分子只能在突触处排列成信号复合物。与紧邻的蛋白质相互作用，这种定位本身可能是由经验调节的。了解功能信号复合物如何产生以及它们如何调节突触因此，强度要求我们探测突触后蛋白质的空间排列传统方法没有足够的分辨率来确定位置。突触蛋白被映射到这些微小（< 1 m）的突触结构中。开发工具来绘制单个突触蛋白（例如谷氨酸）的空间排列受体）内的 PSD，并确定这些空间排列如何受到突触的影响可塑性，使用超分辨率光学显微镜通过绘制许多不同的相对位置。突触后膜内的蛋白质和 PSD 我们将能够生成 3 维模型该方法有望对构成突触后侧的蛋白质晶格进行研究。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结构模型中对其解释至关重要的上下文，并将为突触分析添加一个强大的新工具功能。