Regulation of Serotonin Transporters

血清素转运蛋白的调节

• 批准号: 7988091
• 负责人: Randy D. Blakely
• 金额: $ 31.57万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7988091
• 关键词: Adenosine; Adult; Aggressive behavior; Agonist; Antidepressive Agents; Area; Autistic Disorder; Autoreceptors; Behavior; Behavioral; Biological; Biology; Brain; Brain Diseases; Carrier Proteins; Cocaine; Code; Cognition; Complementary DNA; Complex; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Development; Disease; Drosophila genus; Engineering; Environment; Exhibits; Generations; Homeostasis; Human; Human Cloning; In Vitro; Interleukin-1; Interleukins; Knock-in Mouse; Lead; Link; MAP Kinase Gene; MAPK14 gene; Mediating; Membrane; Mental disorders; Metabolic Clearance Rate; Moods; Mus; Mutation; Neurons; Neurotransmitters; Pathway interactions; Pattern; Pharmaceutical Preparations; Phosphorylation; Phosphotransferases; Physiological; Play; Post-Translational Regulation; Preparation; Protein Kinase; Proteins; Purinergic P1 Receptors; Rattus; Receptor Activation; Regulation; Relative (related person); Research; Rewards; Role; Series; Serotonin; Signal Pathway; Signal Transduction; Synapses; System; Transgenic Mice; Translating; Variant; addiction; base; density; ecstasy; experience; gain of function; human MAPK14 protein; in vivo; link protein; man; mouse model; mutant; novel; novel therapeutic intervention; presynaptic; programs; protein complex; psychostimulant; receptor; response; serotonin transporter; trafficking

DESCRIPTION (provided by applicant): The presynaptic serotonin (5-hydroxytryptamine, 5-HT) transporter (SERT) dictates 5-HT inactivation following release in the brain and periphery and is a major target for antidepressant medications as well as psychostimulants, including cocaine and 3,4-methylenedioxymethamphetamine (MDMA, "Ecstasy"). Studies over the past two decades by our group have progressively illuminated the structural basis for 5-HT clearance and the mechanisms resident within serotonergic neurons and terminals that provide for control of membrane SERT activity, including evidence of trafficking-dependent and independent pathways. Furthermore, we have identified multiple, naturally occurring SERT coding variants (e.g. Gly56Ala) that disrupt SERT regulation in vitro. In the prior iteration of this project, we identified protein kinase G subtype 1 (PKG1) and p38 MAPKa as critical determinants of receptor-driven SERT trafficking and catalytic activation, respectively. In this renewal application, we propose to advance these studies through an integrated series of in vitro, ex vivo and in vivo analyses capitalizing on our increased understanding of receptor systems activating these pathways and the development of novel transgenic mouse models. Through this effort we seek to not only explore the fundamental biology underlying SERT regulation, but also to provide proof of concept for novel therapeutic approaches to treat 5-HT associated brain disorders. In Specific Aim I, we seek to establish the structural basis of SERT regulation by A3A adenosine receptors (A3ARs) and IL-1 receptors (IL-1Rs) that provide for rapid SERT modulation. Specifically, we investigate the hypothesis that these receptors participate in, and signal through physical complexes with SERT and rely on cGMP-dependent SERT phosphorylation as well as altred membrane compartmentation. Finally, we pursue domains and residues that establish the differential control of SERT imparted by these signaling pathways. In Specific Aim II, we elucidate the impact in vitro of naturaly- occuring, human SERT protein variation on receptor- and kinase-mediated SERT regulation and extend our efforts to ask whether coding variation in A3ARs and IL-1Rs mimics the effects of SERT coding variation. In Specific Aim III, we seek to translate our understanding of A3AR, IL-1R and p38 MAPKa pathways to examine the impact on 5-HT homeostasis, 5-HT clearance and synaptic 5-HT signaling in brain preparations. This Aim will make use of both pharmacological approaches as well as a novel line displaying adult, 5-HT-neuron- specific elimination of p38 MAPKa. In Specific Aim IV, we.examine a novel line of mice engineered to expess a disease-associated, hyperphosphorylated coding variant of SERT (SERT Gly56Ala) for effects on phosphorylation, protein complexes, 5-HT clearance and synaptic 5-HT signaling in native preparations. Finally, these mice are explored for impact on behavioral effects of the psychostimulants cocaine and MDMA. PUBLIC HEALTH RELEVANCE: Mental illnesses ranging from autism to addiction are believed to arise from a complex interplay of environment and compromised biological mechanisms that dictate brain development, function and plasticity. The neurotransmitter serotonin has long been understood to play an important, modulatory role in brain signaling and is itself under constant regulatory control. Building from the project leader's experience and expertise in the study of the serotonin transporter (SERT), the brain's chief mechanism for serotonin inactivation and a critical target for antidepressant and addictive drugs, this proposal seeks answers to fundamental questions of SERT regulation and their relationship to synaptic signaling and disease mechanisms.

描述（由申请人提供）：突触前的5-羟色胺（5-羟色胺，5-HT）转运蛋白（SERT）指示大脑和外围释放后5-HT失活，是抗抑郁药以及包括cocaine和3,4-甲基化的抗抑郁药的主要靶标，是抗抑郁药以及精神刺激剂的主要目标。在过去的二十年中，我们小组的研究逐渐阐明了5-HT清除率的结构基础，以及居住在血清素能神经元中的机制和终端，这些机制提供了控制膜SERT活性的控制，包括依赖运输依赖和独立途径的证据。此外，我们已经确定了多种自然存在的SERT编码变体（例如Gly56Ala），它们会在体外破坏SERT调节。在该项目的先前迭代中，我们分别确定了蛋白激酶G亚型1（PKG1）和P38 MAPKA是受体驱动的SERT运输和催化激活的关键决定因素。在此续订应用中，我们建议通过一系列体外，体内和体内分析来推进这些研究，从而利用我们对受体系统的增强理解，从而激活这些途径以及新型转基因小鼠模型的发展。通过这项努力，我们不仅寻求探索SERT调节的基本生物学，而且还寻求为治疗5-HT相关脑部疾病的新型治疗方法提供概念证明。在特定目的I中，我们寻求建立由A3A腺苷受体（A3ARS）和IL-1受体（IL-1R）的SERT调节的结构基础，以提供快速的SERT调节。具体而言，我们研究了这些受体参与并通过SERT的物理复合物参与的假设，并依赖于CGMP依赖性的SERT磷酸化以及疏忽的膜隔室。最后，我们追求建立由这些信号通路赋予SERT的差分控制的域和残基。在特定的目标II中，我们阐明了自然发生的体外影响，人类SERT蛋白质变异对受体和激酶介导的SERT调节，并扩大了我们询问A3ARS和IL-1RS中编码变化是否模仿SERT编码变化的效果。在特定的目标III中，我们试图翻译我们对A3AR，IL-1R和P38 MAPKA途径的理解，以检查对5-HT稳态，5-HT清除率和突触5-HT信号的影响。这个目标将利用两种药理学方法以及显示成人5-ht-neuron-特异性消除p38 MAPKA的新系列。在特定的目标IV中，我们将一组新型小鼠系进行了设计，该系列旨在调查SERT（SERT GLY56ALA）的疾病相关的，高磷酸化的编码变体，以对天然制剂中的磷酸化，蛋白质复合物，5-HT清除率和突触5-HT信号的影响。最后，探索了这些小鼠，以影响可卡因和MDMA的心理刺激剂的行为影响。 公共卫生相关性：从自闭症到成瘾的精神疾病源于环境的复杂相互作用以及决定大脑发育，功能和可塑性的生物学机制。长期以来，神经递质5-羟色胺在脑信号传导中起着重要的调节作用，并且本身在恒定的调节控制下。该提案从项目领导者的经验和专业知识（SERT）研究，大脑灭活的主要机制以及抗抑郁药和成瘾性药物的关键目标中建立，该提案为SERT调节的基本问题及其对突触信号和疾病机制的关系寻求答案。