Molecular Mechanisms Underlying PSD-MAGUK/NMDA Receptor Interactions

PSD-MAGUK/NMDA 受体相互作用的分子机制

基本信息

批准号：
8245101
负责人：
MacKenzie A Howard
金额：
$ 5.57万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-01 至 2013-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8245101
关键词：
AMPA Receptors Alzheimer&apos s Disease Amino Acid Motifs Anatomy Autistic Disorder Brain Cells DLG1 gene Degenerative Disorder Dendrites Development Electrophysiology (science)Future Genetic Glutamate Receptor Glutamates Goals Image Individual Kinetics Knock-out Knockout Mice Learning Long-Term Potentiation Measures Mediating Memory Mental Retardation Mental disorders Molecular Molecular Genetics Morphology N-Methyl-D-Aspartate Receptors Neuraxis Neurons Nomenclature Pattern Pharmacology Physiology Play Population Protein Binding Protein Binding Domain Protein Family Proteins Research Role Scaffolding Protein Schizophrenia Scientist Shapes Signal Transduction Synapses Synaptic Transmission Synaptic plasticity System Techniques Tertiary Protein Structure Testing base design developmental disease discs, large (Drosophila) homolog 2 protein, rat in vivo membrane-associated guanylate kinase nervous system disorder overexpression presynaptic density protein 95 receptor relating to nervous system research study synapse-associated protein 97 trafficking transmission process

项目摘要

DESCRIPTION (provided by applicant): The overall objectives of my proposal are to understand the molecular mechanisms by which glutamate receptors, particularly NMDA receptors (NMDARs), interact with synaptic scaffolding proteins and how these interactions shape synaptic transmission. Specifically, I will study how the postsynaptic density-95-like membrane associated guanylate kinase (PSD-MAGUK) protein family, in particular the protein SAP97, traffic NMDA receptors subunits and change their physiology. PSD-MAGUKs and NMDA receptors play a critical role in basal synaptic transmission and learning and memory, and have been implicated in a wide variety of neurological diseases, ranging from developmental disorders such as autism, schizophrenia, to degenerative diseases such as Alzheimer's. My research goals are outlined in two Specific Aims: Specific Aim 1: SAP97 controls AMPA and NMDA receptor trafficking and synaptic morphology. I hypothesize that SAP97 traffics AMPA and NMDARs to synapses during early development and specifically promotes GluN2A-containing NMDARs. Second, I hypothesize that SAP97-mediated signaling also controls dendrite and synapse morphology in developing neurons. I will manipulate SAP97 protein levels in vivo and use electrophysiology and confocal imaging to measure the role of this protein in synaptic transmission and neuronal anatomy. Specific Aim 2: Molecular differences in PSD-MAGUKs underlie NMDAR kinetics and subunit switching. First, I hypothesize that specific protein binding domains shared by PSD-93, -95, and SAP97 promote synaptic trafficking of GluN2A-containing NMDARs while different motifs in SAP102 promote GluN2B- containing receptors. Second, I hypothesize that PSD-MAGUKs also directly influence NMDAR physiology, with each PSD-MAGUK differentially interacting with NMDARs and shaping synaptic currents. I will design and overexpress chimeric PSD-MAGUK proteins in vivo, in NMDAR subunit conditional knockout mice, and measure the effect on NMDARs using electrophysiology. I will also use a heterologous expression system to measure direct interactions between these proteins. Thus, I will define the protein domains responsible for PSD-MAGUK/NMDAR interactions and how these interactions alter NMDAR physiology. These experiments take a multi-dimensional approach to a vital scientific question, combining cutting edge molecular genetic, physiologocial, and anatomical techniques and will enhance our understanding of fundamental molecular mechanisms of synaptic transmission and learning and memory.

描述（由申请人提供）：我的提案的总体目标是了解谷氨酸受体，特别是 NMDA 受体（NMDAR）与突触支架蛋白相互作用的分子机制，以及这些相互作用如何影响突触传递。具体来说，我将研究突触后密度 95 样膜相关鸟苷酸激酶 (PSD-MAGUK) 蛋白家族，特别是 SAP97 蛋白，如何运输 NMDA 受体亚基并改变其生理机能。 PSD-MAGUK 和 NMDA 受体在基础突触传递以及学习和记忆中发挥着关键作用，并且与多种神经系统疾病有关，从自闭症、精神分裂症等发育障碍到阿尔茨海默氏症等退行性疾病。我的研究目标概括为两个具体目标：具体目标 1：SAP97 控制 AMPA 和 NMDA 受体运输和突触形态。我假设 SAP97 在早期发育过程中将 AMPA 和 NMDAR 运输到突触，并专门促进含有 GluN2A 的 NMDAR。其次，我假设 SAP97 介导的信号传导也控制发育中神经元的树突和突触形态。我将操纵体内 SAP97 蛋白水平，并使用电生理学和共聚焦成像来测量该蛋白在突触传递和神经元解剖学中的作用。具体目标 2：PSD-MAGUK 的分子差异是 NMDAR 动力学和亚基转换的基础。首先，我假设 PSD-93、-95 和 SAP97 共享的特定蛋白结合域促进含有 GluN2A 的 NMDAR 的突触运输，而 SAP102 中的不同基序则促进含有 GluN2B 的受体。其次，我假设 PSD-MAGUK 也直接影响 NMDAR 生理学，每个 PSD-MAGUK 与 NMDAR 的相互作用不同，并塑造突触电流。我将在 NMDAR 亚基条件敲除小鼠体内设计并过表达嵌合 PSD-MAGUK 蛋白，并使用电生理学测量对 NMDAR 的影响。我还将使用异源表达系统来测量这些蛋白质之间的直接相互作用。因此，我将定义负责 PSD-MAGUK/NMDAR 相互作用的蛋白质结构域以及这些相互作用如何改变 NMDAR 生理学。这些实验采用多维方法解决重要的科学问题，结合尖端的分子遗传学、生理学和解剖学技术，将增强我们对突触传递以及学习和记忆的基本分子机制的理解。