Amphetamine Regulation of Dopamine Transport

安非他明对多巴胺运输的调节

• 批准号: 7655415
• 负责人: AURELIO GALLI
• 金额: $ 29.35万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7655415
• 关键词: Acute; Address; Alanine; Amphetamines; Antidepressive Agents; Aspartate; Behavioral; Biochemical; Cell membrane; Cocaine; Cognitive; Disease; Dopamine; Drug Addiction; Emotional; Goals; Grant; Image; Indium; Link; Mediating; Membrane Proteins; Methodology; Mitogen-Activated Protein Kinases; Modeling; Modification; Molecular; Molecular Target; Mutation; N-terminal; Neurons; Neurotransmitters; Norepinephrine; Parkinson Disease; Pathway interactions; Pharmaceutical Preparations; Phosphorylation; Phosphotransferases; Preparation; Property; Proteins; Proto-Oncogene Proteins c-akt; Regulation; Research; Research Personnel; Resistance; Rewards; Role; SNAP receptor; Schizophrenia; Serine; Signal Transduction; Simulate; Specificity; Synapses; System; Testing; Time; Translating; Work; addiction; base; calmodulin-dependent protein kinase II; dopamine transporter; extracellular; millisecond; nervous system disorder; novel; programs; psychostimulant; research study; reuptake; serotonin transporter; stimulant abuse; syntaxin 1A; trafficking; uptake

DESCRIPTION (provided by applicant): The long-term objective of this project is to develop an understanding of the acute modulation of dopamine (DA) transporter (DAT) function by amphetamine (AMPH) and other DAT substrates. DA signaling at CNS synapses regulates a variety of cognitive, emotional and behavioral functions. Abnormalities in the dopaminergic system have been implicated in a number of psychiatric and neurological disorders, including drug addiction, schizophrenia and Parkinson's disease. AMPH induces reverse transport of DA, thereby increasing extracellular DA levels and leading to its psychostimulant effects. Exciting new experiments suggest that AMPH has unique interactions with the DAT. Thus, in contrast to DA, AMPH stimulates CaMKII activity, causes DAT associations with the SNARE protein syntaxin 1A, and induces DAT channel-like activity that results in rapid (msec) bursts of DA efflux. This project will combine molecular and biochemical approaches with biophysical and real-time imaging methodologies to characterize AMPH regulation of DAT function both in heterologous and neuronal preparations. The key issues to resolve include how AMPH differs from DA in its ability to induce DAT-mediated reverse transport of DA and DAT channel-like activity and whether DAT N-terminal phosphorylation and subsequent protein associations are essential for these actions of AMPH. Our experimental plan links the AMPH-induced functional regulation of DAT to stimulation of cytoplasmic signals that regulate the association of DAT and other proteins, thereby modulating DA efflux. The proposed studies address the following Specific Aims: 1) To identify differences in the abilities of AMPH and other DAT substrates to stimulate cytoplasmic signals leading to DA efflux. 2) To identify elements in the N-terminus that regulate the action of AMPH and to assess the impact of phosphorylation on the ability of AMPH to induce DA efflux. 3) To determine whether kinase activity and phosphorylation of DAT N-terminal serines regulate the interaction between DAT and syntaxin 1A, thereby modulating AMPH-induced DA efflux. The goal is to generate an experimentally-based model advancing our understanding of acute AMPH regulation of the DAT and to discover novel pathways and molecules that may contribute to disrupted dopaminergic signaling in disease states such as addiction.

描述（由申请人提供）：该项目的长期目标是建立对苯丙胺（AMPH）和其他DAT底物的多巴胺（DA）转运蛋白（DAT）功能的急性调制的理解。中枢神经系统的DA信号传导调节多种认知，情感和行为功能。多巴胺能系统的异常已与许多精神病和神经系统疾病有关，包括吸毒成瘾，精神分裂症和帕金森氏病。 AMPH诱导了DA的反向运输，从而提高了细胞外DA水平并导致其精神刺激作用。令人兴奋的新实验表明，AMPH与DAT具有独特的相互作用。因此，与DA相反，AMPH刺激CAMKII活性，引起与SNARE蛋白语法1a的DAT关联，并诱导DAT通道样活性，从而导致DA外排的快速（MSEC）爆发。该项目将将分子和生化方法与生物物理和实时成像方法相结合，以表征异源和神经元制剂中DAT功能的AMPH调节。解决的关键问题包括AMPH与DA诱导DAT介导的DA和DAT通道样活动的反向转运的能力以及DAT N末端磷酸化和随后的蛋白质关联是否对于AMPH的这些作用至关重要。我们的实验计划将AMPH诱导的DAT功能调节与调节DAT和其他蛋白质缔合的细胞质信号的刺激联系起来，从而调节DA外排。拟议的研究涉及以下特定目的：1）确定AMPH和其他DAT底物刺激导致DA外流的胞质信号的能力差异。 2）确定N末端中调节AMPH作用并评估磷酸化对AMPH诱导DA外排能力的影响的要素。 3）确定DAT N末端丝氨酸的激酶活性和磷酸化是否调节DAT和语法1a之间的相互作用，从而调节Amph诱导的DA外排。目的是生成一个基于实验的模型，以促进我们对DAT的急性AMPH调节的理解，并发现新的途径和分子，这些途径和分子可能导致疾病状态（例如成瘾状态）中断的多巴胺能信号传导。