Signaling Scaffold of NMDA Receptor-Dependent Long-Term Plasticity

NMDA 受体依赖性长期可塑性的信号支架

• 批准号: 7244564
• 负责人: WEIFENG XU
• 金额: $ 9万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-05 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7244564
• 关键词: A kinase anchoring protein; AMPA Receptors; Acute; Address; Alzheimer' s Disease; Binding; Calcineurin; Cells; Chromosome Pairing; Cyclic AMP-Dependent Protein Kinases; DLG1 gene; DLG4 gene; Data; Dependence; Development; Epilepsy; Family member; Gene Silencing; Gene Transfer; Genes; Glutamate Receptor; Glutamates; Hippocampus (Brain); Learning; Lentivirus Vector; Long-Term Depression; Long-Term Potentiation; Mediating; Memory; Mental Depression; Mental disorders; Mentors; Molecular; N-Methyl-D-Aspartate Receptors; N-terminal; Neurologic; Neurons; Numbers; Phase; Play; Property; Protein Family; Proteins; Research; Role; Scaffolding Protein; Schizophrenia; Series; Signal Transduction; Signaling Molecule; Signaling Protein; Site; Slice; Specificity; Subfamily lentivirinae; Synapses; Synaptic plasticity; System; Testing; Thinking; density; experience; genetic regulatory protein; lentiviral-mediated; membrane-associated guanylate kinase; mutant; neural circuit; patch clamp; postsynaptic; presynaptic density protein 95; receptor function; research study; scaffold; small hairpin RNA; stargazin; synaptic function; trafficking; transmission process

DESCRIPTION (provided by applicant): Activity-dependent changes in synaptic strength at glutamatergic synapses are thought to contribute to the development of neural circuitry and many forms of experience-dependent plasticity, including learning and memory. The hippocampus, a major site of synaptic plasticity, plays a fundamental role in some forms of learning and memory, and has been implicated in a number of neurological and psychiatric disorders, including depression, epilepsy, Alzheimer's disease, and schizophrenia. In this application, I outline a series of experiments that will test the functional significance of key synaptic scaffolding proteins in regulating glutamate receptor function and synaptic plasticity at hippocampal Schaffer collateral-CA1 synapses. This will involve making simultaneous whole cell patch clamp recordings from neurons in organotypic hippocampal slice cultures that haven been molecularly modified using lentiviral-mediated gene knockdown via shRNA and simultaneous lentiviral-mediated gene transfer. The bicistronic lentiviral vector I will use allows expression of mutant forms of a protein on the background of acute knockdown of the endogenous protein. I will specifically focus on the function of the postsynaptic scaffolding proteins of the family of the disc-large (DLG) membrane-associated guanylate kinases (MAGUKs) and their interacting partners. My previous results demonstrate that two family members of DLG-MAGUKs, PSD-95 and SAP97, regulate synaptic AMPAR function differently in terms of their activity-dependence. My preliminary studies also show that the effects of PSD-95 on basal transmission and long-term depression are dissociable. The specific objectives of this application are: (1) to analyze the signaling scaffold that is important for long-term depression (LTD), in particular, the role of specific domains in PSD-95 and the A-kinase anchoring protein 79/150 (AKAP79/150), and (2) to examine the interaction of PSD-95 with transmembrane AMPA receptor regulatory proteins (TARPs) in mediating long-term potentiation (LTP). Results from these experiments will begin to elucidate how dynamic interactions among different components of the postsynaptic density influence synaptic function and will address fundamental questions about how signaling specificity is achieved during different forms of synaptic plasticity.

描述（由申请人提供）：谷氨酸能突触的突触强度的活动依赖性变化被认为有助于神经回路和许多形式的经验依赖性可塑性的发育，包括学习和记忆。海马是突触可塑性的主要部位，在某些形式的学习和记忆中起着重要作用，并且与许多神经和精神疾病有关，包括抑郁症、癫痫、阿尔茨海默病和精神分裂症。在这个应用程序中，我概述了一系列的实验，将测试在调节谷氨酸受体功能和突触可塑性在海马Schaffer侧支-CA 1突触的关键突触支架蛋白的功能意义。这将涉及从器官型海马切片培养物中的神经元同时进行全细胞膜片钳记录，所述培养物已经使用慢病毒介导的基因敲低通过shRNA和同时的慢病毒介导的基因转移进行分子修饰。我将使用的双顺反子慢病毒载体允许在内源性蛋白质的急性敲低的背景下表达蛋白质的突变形式。我将特别关注大椎间盘（DLG）膜相关鸟苷酸激酶（MAGUKs）家族的突触后支架蛋白及其相互作用伙伴的功能。我以前的研究结果表明，DLG-MAGUKs的两个家族成员，PSD-95和SAP 97，调节突触AMPAR功能不同的活动依赖性。我的初步研究还表明，PSD-95对基础传输和长期抑郁的影响是分离的。本申请的具体目标是：（1）分析对长期抑郁症（LTD）重要的信号传导支架，特别是PSD-95和A激酶锚定蛋白79/150中特定结构域的作用（AKAP 79/150），（2）研究PSD-95与跨膜AMPA受体调节蛋白（TARP）在长时程增强（LTP）中的相互作用。这些实验的结果将开始阐明突触后密度的不同组成部分之间的动态相互作用如何影响突触功能，并将解决关于在不同形式的突触可塑性期间如何实现信号特异性的基本问题。