Presynaptic Regulation of C.elegans Dopamine Transporter

线虫多巴胺转运蛋白的突触前调节

• 批准号: 8641780
• 负责人: Randy D. Blakely
• 金额: $ 12.93万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-15 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8641780
• 关键词: Alleles; Amphetamines; Animal Model; Animals; Attention; Attention Deficit Disorder; Attention deficit hyperactivity disorder; Behavioral; Behavioral Assay; Biochemical; Biological Assay; Biological Models; Bipolar Disorder; Brain; Caenorhabditis elegans; Carrier Proteins; Cocaine; Code; Cognition; Cognitive deficits; Coupling; Data; Development; Disease; Dopamine; Dopamine Receptor; Double-Stranded RNA; Elements; Evaluation; Exhibits; G protein coupled receptor kinase; G-substrate; Gene Expression; Genes; Genetic; Genetic Screening; Genomics; Homeostasis; Homologous Gene; Human; Hyperactive behavior; Impulsivity; In Vitro; Induced Mutation; Injection of therapeutic agent; Liquid substance; Maintenance; Mammals; Maps; Mediating; Membrane Microdomains; Methylphenidate; Modeling; Motor Activity; Movement; Mus; Mutation; Nematoda; Neurotransmitters; Orthologous Gene; Paralysed; Parkinson Disease; Pharmaceutical Preparations; Phenotype; Phosphoric Monoester Hydrolases; Photobleaching; Phylogeny; Physiological; Probability; Production; Receptor Signaling; Recovery; Regulation; Reporter; Research; Reserpine; Rewards; Ritalin; Schizophrenia; Sequence Alignment; Signal Transduction; Signaling Protein; Site; Solid; Surface; Swimming; Synapses; Techniques; Testing; Therapeutic; Transfection; Transgenic Organisms; Variant; Vesicle; Water; addiction; dopamine transporter; dopaminergic neuron; executive function; extracellular; feeding; genetic manipulation; in vivo; insight; man; mutant; novel; optogenetics; positional cloning; presynaptic; protein expression; psychostimulant; research study; response; reuptake; skills; theories; therapeutic target; trafficking; translational study; uptake; vesicle-associated membrane protein

DESCRIPTION (provided by applicant): Dopamine (DA) is a neurotransmitter used throughout phylogeny to modulate circuits controlling movement, attention, reward and cognition. Alterations in DA signaling in man have been implicated in Parkinson's Disease, Attention-Deficit Hyperactivity Disorder (ADHD), addiction and schizophrenia. DA signaling is tightly controlled by a presynaptic, DA transporter (DAT) that is a major target for addictive psychostimulants such as cocaine and amphetamine (AMPH), as well as agents used in the treatment of ADHD. Our recent identification of multiple, functional alleles of human DAT in ADHD and Bipolar disorder subjects adds translational significance to our efforts to decipher the presynaptic mechanisms that control DA release and inactivation. To date, these mechanisms have largely been studies through pharmacological and genetic manipulation of genes identified two decades or more ago. In the current application, we capitalize on a robust, forward genetic approach to identify and characterize novel presynaptic regulators of DA signaling using the powerful model system Caenorhabditis elegans. Over the past 15 years, the Blakely lab has developed skills in the C. elegans model with a focus on DAT and DA signaling, initiated by the identification of the C. elegans DAT gene (T23G5.5, dat-1). The present effort arises from our discovery of a simple DA and dat-1 dependent phenotype termed Swimming-Induced Paralysis (SWIP). Whereas wildtype animals thrash in water at ~1 Hz for up to 30 minutes, dat-1 (ok157) (DAT-deficient) animals paralyze in 3-5 minutes. This phenotype is dependent on DA synthesis, vesicular DA packaging, and DA release and is overcome by DAT activity. We request support to advance our screen to identify and characterize genes acting presynaptically to regulate DA release and reuptake, localize their mode of contribution to DA signaling, assess their impact on the actions of AMPH, and initiate an analysis of conserved vertebrate homologs. Together our efforts provide an opportunity to identify novel and conserved regulators of DA signaling that would be extremely difficult to elucidate in vertebrate models.

描述（由申请人提供）：多巴胺（DA）是整个系统发育中使用的神经递质，用于调节控制运动，注意力，奖励和认知的电路。人类发信号的改变与帕金森氏病，注意力缺陷多动症（ADHD），成瘾和精神分裂症有关。 DA信号传导由突触前的DA转运蛋白（DAT）严格控制，该转运蛋白是可卡因和苯丙胺（AMPH）等成瘾性心理刺激剂的主要靶标，以及用于治疗ADHD的药物。我们最近对ADHD和躁郁症受试者中人类DAT的多种功能等位基因的鉴定为我们破译控制DA释放和失活的突触前机制的努力增添了转化意义。迄今为止，这些机制在很大程度上是通过对识别二十年或更长时间的基因的药理和遗传操纵进行研究的。在当前的应用中，我们利用强大的模型系统Caenorhabditis秀丽隐杆线虫来识别和表征新型DA信号的新型DA信号调节剂。在过去的15年中，Blakely Lab在秀丽隐杆线虫模型中开发了技能，专注于DAT和DA信号，这是由C. C. elegans dat Gene（T23G5.5，DAT-1）启动的。目前的努力是由于我们发现了一个简单的DA和Dat-1依赖性表型，称为游泳引起的瘫痪（SWIP）。而野生型动物在〜1 Hz的水中最多30分钟，dat-1（OK157）（缺乏DAT缺乏）动物在3-5分钟内瘫痪。该表型取决于DA合成，囊泡DA包装和DA释放，并通过DAT活动克服。我们要求提供支持以推进屏幕，以识别和表征在突触前作用以调节DA释放和重新摄取，定位其对DA信号传导的贡献方式，评估其对AMPH作用的影响，并启动保守脊椎动物同源物的分析。我们的努力共同提供了一个机会来识别DA信号传导的新颖和保守的调节剂，这在脊椎动物模型中很难阐明。