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: 7996638
• 负责人: WEIFENG XU
• 金额: $ 24.28万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 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 2012-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7996638
• 关键词: A kinase anchoring protein; AMPA Receptors; Acute; Address; Alzheimer' s Disease; Binding; Brain; Brain Diseases; C-terminal; Calcineurin; Cells; Cyclic AMP-Dependent Protein Kinases; DLG1 gene; Dependence; Development; Dimerization; 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; Molecular; N-Methyl-D-Aspartate Receptors; N-terminal; National Institute of Mental Health; Neurons; Play; Process; Protein Family; Proteins; Research; Role; Scaffolding Protein; Schizophrenia; Series; Signal Transduction; Signaling Molecule; Signaling Protein; Site; Slice; Specificity; Structure; Subfamily lentivirinae; Synapses; Synaptic plasticity; Testing; Training; density; experience; falls; genetic regulatory protein; lentiviral-mediated; membrane-associated guanylate kinase; mutant; nervous system disorder; neural circuit; patch clamp; postsynaptic; receptor function; research study; scaffold; small hairpin RNA; stargazin; synaptic function; trafficking; transmission process

Activity-dependent ciianges in synaptic strengtli at glutamatergic synapses are tiiougiit to contribute to tiie 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 proposal, 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-CAl 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. During the K99 training period, I found that the effects of PSD-95 on basal transmission and long-term depression are dissociable. The N-terminal domain of PSD-95 is required for dimerization and appropriate synaptic enrichment of PSD-95 but alone does not influence synaptic function. The C-terminal portion of PSD-95 serves a dual function. It is required to localize PSD-95 at the synapse and as a scaffold for critical downstream signaling proteins that are required for LTD. The specific objectives of my independent research are: (1) to analyze the signaling scaffold that is important for long-term depression (LTD), in particular, the role of 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), (3) to examine the role of SAP97 in mediating 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

谷氨酸能突触的突触强度中的活动依赖性变化可能有助于神经元的发育 神经回路和许多形式的依赖于经验的可塑性，包括学习和记忆。这 海马体是突触可塑性的主要部位，在某些形式的学习和记忆中发挥着基础作用，并且 与许多神经和精神疾病有关，包括抑郁症、癫痫症、阿尔茨海默氏症 疾病和精神分裂症。在这个提案中，我概述了一系列实验，这些实验将测试以下功能的意义： 调节海马谷氨酸受体功能和突触可塑性的关键突触支架蛋白 Schaffer 侧支-CA1 突触。这将涉及同时进行全细胞膜片钳记录 使用慢病毒介导的基因对器官型海马切片培养物中的神经元进行分子修饰 通过 shRNA 和同时慢病毒介导的基因转移进行敲除。我将使用的双顺反子慢病毒载体 允许在内源蛋白急性敲低的背景下表达突变形式的蛋白。我会 特别关注椎间盘大（DLG）家族突触后支架蛋白的功能 膜相关鸟苷酸激酶 (MAGUK) 及其相互作用的伙伴。我之前的结果表明 DLG-MAGUK 的两个家族成员 PSD-95 和 SAP97 在 就他们的活动依赖性而言。在K99训练期间，我发现PSD-95对基础的影响 传播和长期抑郁是分不开的。二聚化需要 PSD-95 的 N 端结构域 PSD-95 的适当突触富集但单独使用并不影响突触功能。 C端部分 PSD-95 具有双重功能。需要将 PSD-95 定位在突触处并作为关键的支架 LTD 所需的下游信号蛋白。我独立研究的具体目标是：（1） 分析对长期抑郁 (LTD) 很重要的信号支架，特别是 A-激酶的作用 锚定蛋白 79/150 (AKAP79/150)，以及 (2) 检查 PSD-95 与跨膜 AMPA 的相互作用 受体调节蛋白 (TARP) 在介导长时程增强 (LTP) 中的作用，(3) 检查 SAP97 在调节长时程增强 (LTP) 中的作用 介导 LTP。这些实验的结果将开始阐明不同类型之间的动态相互作用 突触后密度的组成部分影响突触功能，并将解决有关如何突触的基本问题 信号传导特异性是在不同形式的突触可塑性期间实现的

项目成果

PSD-95-like membrane associated guanylate kinases (PSD-MAGUKs) and synaptic plasticity.

• DOI: 10.1016/j.conb.2011.03.001
• 发表时间: 2011-04
• 期刊: Current opinion in neurobiology
• 影响因子: 5.7
• 作者: Xu W

Shank Proteins Differentially Regulate Synaptic Transmission.

• DOI: 10.1523/eneuro.0163-15.2017
• 发表时间: 2017-11
• 期刊: eNeuro
• 影响因子: 3.4
• 作者: Shi R;Redman P;Ghose D;Hwang H;Liu Y;Ren X;Ding LJ;Liu M;Jones KJ;Xu W
• 通讯作者: Xu W

Durable fear memories require PSD-95.

• DOI: 10.1038/mp.2014.161
• 发表时间: 2015-07
• 期刊: MOLECULAR PSYCHIATRY
• 影响因子: 11
• 作者: Fitzgerald, P. J.;Pinard, C. R.;Camp, M. C.;Feyder, M.;Sah, A.;Bergstrom, H. C.;Graybeal, C.;Liu, Y.;Schlueter, O. M.;Grant, S. G.;Singewald, N.;Xu, W.;Holmes, A.
• 通讯作者: Holmes, A.