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）：我的建议的总体目标是了解分子机制，谷氨酸受体，尤其是NMDA受体（NMDAR），与突触型支架蛋白相互作用以及这些相互作用如何形成突触传播。具体而言，我将研究突触后密度95样膜相关的鸟苷酸激酶（PSD-Maguk）蛋白质家族如何，尤其是蛋白质SAP97，交通NMDA受体亚基并改变其生理学。 PSD-Maguks和NMDA受体在基础突触传播，学习和记忆中起着至关重要的作用，并且与各种神经系统疾病有关，从自闭症，精神分裂症等发育障碍到诸如阿尔茨海默氏症等退化性疾病。我的研究目标在两个具体目标中概述：特定目标1：SAP97控制AMPA和NMDA受体运输和突触形态。我假设SAP97流量AMPA和NMDAR在早期开发过程中会突触，并专门促进了含Glun2a的NMDAR。其次，我假设SAP97介导的信号传导还控制着发育中的神经元中的树突和突触形态。我将在体内操纵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播和神经元解剖结构中的作用。特定目标2：PSD-Maguks的分子差异基础NMDAR动力学和亚基转换。首先，我假设由PSD-93，-95和SAP97共享的特定蛋白结合结构域促进了含Glun2a的NMDAR的突触运输，而SAP102中的不同基序则促进了含有Glun2b的受体。其次，我假设PSD-Maguks也直接影响NMDAR生理学，每个PSD-Maguk都与NMDARs差异相互作用并塑造突触电流。我将在体内设计和过表达嵌合PSD-maguk蛋白，在NMDAR亚基有条件敲除小鼠中，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来解决一个重要的科学问题，结合了尖端分子遗传，生理社会和解剖技术，并将增强我们对突触传播以及学习和记忆和记忆和记忆和记忆和记忆和记忆和记忆和记忆和记忆基本机制的理解。