The Molecular Mechanisms of the GDNF-mediated Decrease in Alcohol Consumption

GDNF 介导的减少饮酒的分子机制

• 批准号: 7695562
• 负责人: Somayeh Ahmadiantehrani
• 金额: $ 3.01万
• 依托单位: ERNEST GALLO CLINIC AND RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7695562
• 关键词: Accounting; Acute; Affect; Alcohol consumption; Alcohol dependence; Alcoholism; Alcohols; Behavior Control; Behavioral; Biochemical; Biological Models; Brain; Brain-Derived Neurotrophic Factor; Cell Line; Chronic; Consumption; Data; Development; Disease; Dopamine; Dose; Drug Addiction; Esthesia; Ethanol; Exposure to; Extracellular Signal Regulated Kinases; Figs - dietary; Growth Factor; High Pressure Liquid Chromatography; Hour; In Vitro; Individual; Infusion procedures; Laboratories; Lead; Life; MEKs; Mediating; Midbrain structure; Molecular; Molecular Target; Motivation; Neurobiology; Neurons; Neurotransmitters; Nuclear; Nucleus Accumbens; Pathway interactions; Pharmaceutical Preparations; Pharmacotherapy; Phosphorylation; Phosphotransferases; Play; Potassium; Potassium Channel; Preparation; Proteins; Rattus; Regulation; Research; Reverse Transcriptase Polymerase Chain Reaction; Rewards; Rodent; Role; Self Administration; Signal Pathway; Signal Transduction; Small Interfering RNA; Synapsin I; Synapsins; Synaptic Transmission; Synaptic Vesicles; Techniques; Testing; Time; U-0126; Up-Regulation; Ventral Tegmental Area; Work; alcohol effect; alcohol exposure; cocaine exposure; dopaminergic neuron; drinking; experience; glial cell-line derived neurotrophic factor; immortalized cell; in vitro Model; in vivo Model; inhibitor/antagonist; knowledge base; neuronal growth; neurotransmitter release; neurotrophic factor; novel; problem drinker; receptor

DESCRIPTION (provided by applicant): Compulsive consumption of alcohol is a disease state that affects otherwise healthy individuals. The GDNF signaling pathway plays a role in the regulation of alcohol consumption. The proposed research has two broad aims: 1) to determine the downstream effectors and molecular mechanisms that underlie this action of GDNF, and 2) to determine whether the GDNF signaling pathway is part of a homeostatic mechanism by which alcohol consumption is negatively regulated. Firstly, the actions of GDNF signaling on the phosphorylation of two candidate proteins, Kv4.3 and Synapsin I, will be investigated. Use of in vitro model systems, including an immortalized cell line and primary neuron cultures, will be used to biochemically determine the extent of protein phosphorylation. Because the neurotransmitter dopamine plays a major role in the consumption of alcohol, the effects of GDNF, as well as the GDNF-mediated phosphorylation of these protein targets, on dopamine release will be investigated. siRNA knockdown of the Kv4.3 and/or Synapsin I proteins, along with high-performance liquid chromatography (HPLC) to quantify dopamine release, will be used to determine whether either candidate effector functions in GDNF-mediated dopamine release. Secondly, the effect of ethanol on the upregulation of GDNF and subsequent activation of its signaling pathway will be further characterized in both in vitro and in vivo models. RT-PCR and biochemical techniques will be used to support this aim. Understanding the effect of alcohol consumption on GDNF signaling, as well as the effects of GDNF on alcohol consumption, will contribute to the understanding of the neurobiology of alcohol addiction, and the development of efficacious therapies to alleviate the alcoholic's disease state. Relevance: Alcoholism is a widespread and costly societal problem. Identifying novel molecular targets that may be used to develop drug therapies that can regulate alcohol consumption is a worthwhile pursuit. This research will contribute to the current knowledge base for the understanding and treatment of this and other drug addictions.

描述(申请人提供)：强迫性饮酒是一种疾病状态，影响其他健康的个人。GDNF信号通路在酒精消耗的调节中起着重要作用。这项拟议的研究有两个广泛的目标：1)确定GDNF这一作用的下游效应器和分子机制；2)确定GDNF信号通路是否是酒精消耗负面调节的动态平衡机制的一部分。首先，我们将研究GDNF信号在Kv4.3和突触素I两个候选蛋白磷酸化过程中的作用。使用体外模型系统，包括永生化细胞系和原代神经元培养，将被用于生化确定蛋白质磷酸化的程度。由于神经递质多巴胺在酒精消费中起主要作用，因此将研究GDNF以及GDNF介导的这些蛋白质靶标的磷酸化对多巴胺释放的影响。Kv4.3和/或突触素I蛋白的siRNA敲除，以及用于定量多巴胺释放的高效液相色谱(HPLC)，将被用来确定这两个候选效应器是否在GDNF介导的多巴胺释放中发挥作用。其次，乙醇对GDNF上调及其信号通路激活的影响将在体外和体内模型中得到进一步表征。将使用RT-PCR和生化技术来支持这一目标。了解饮酒对GDNF信号的影响，以及GDNF对饮酒的影响，将有助于了解酒精成瘾的神经生物学机制，并开发有效的治疗方法来缓解酗酒者的疾病状态。相关性：酗酒是一个广泛存在且代价高昂的社会问题。寻找新的分子靶点可以用来开发可以调节酒精消费的药物疗法，这是一个值得追求的目标。这项研究将有助于目前了解和治疗这种和其他药物成瘾的知识基础。