Role of BK Channel Interactome in Excessive Ethanol Drinking

BK 通道相互作用组在过量乙醇饮酒中的作用

基本信息

批准号：
8231180
负责人：
Candice Contet
金额：
$ 26.76万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-05 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8231180
关键词：
Address Affect Alcohol abuse Alcohol consumption Alcohol dependence Alcoholic Intoxication Alcohols Amygdaloid structure Animals Behavior Behavioral Biological Assay Brain Brain region Calcium Candidate Disease Gene Chronic Collaborations Dependence Development Ethanol Ethanol dependence Frequencies Genes Genetic Determinism Health Heavy Drinking Intoxication Investigation Knockout Mice Maps Measures Mediating Meta-Analysis Molecular Molecular Target Neurons Nucleus Accumbens Phenotype Physiology Population Potassium Potassium Channel Prefrontal Cortex Proteins Research Research Personnel Research Project Grants Role Sampling Self Administration Shapes Synapses Synaptic plasticity Testing Transcript Withdrawal Work alcohol effect alcohol exposure alcoholism therapy cost drinking in vivo innovation insight interest large-conductance calcium-activated potassium channels mouse model neurotransmission neurotransmitter release novel preference protein expression research study response

项目摘要

DESCRIPTION (provided by applicant): Alcohol abuse and dependence affect an estimated 8.5% of the U.S. population and are responsible for substantial health and societal costs. The large conductance calcium-activated potassium (BK) channel, together with more than 20% of the proteins known to interact with BK a subunit, have been identified by the INIA-West consortium as potential genetic determinants of ethanol preference. In the present proposal, we hypothesize that perturbation of BK channel interaction network by alcohol in relevant brain regions contributes to excessive drinking. We will focus our investigation on the neurocircuitry subtending the motivational effects of ethanol (nucleus accumbens, amygdala and prefrontal cortex). We further hypothesize that ethanol-induced reorganization of the BK channel interactome mediates changes in neurotransmission and synaptic plasticity observed in ethanol-dependent animals. To test these hypotheses, we will quantify and manipulate expression levels of key interaction partners of the BK a subunit. The first Specific Aim is to characterize the phenotype of knockout mice deficient for either of the two neuronal auxiliary subunits of the BK channel. We will use assays of ethanol intoxication, tolerance and withdrawal, as well as paradigms of voluntary drinking leading to moderate or excessive ethanol intake. The second Specific Aim is to map the expression of known BK interaction partners in the nucleus accumbens, amygdala and prefrontal cortex, and assess how excessive ethanol exposure alters their protein levels. An innovative protein assay will be exploited for the simultaneous quantification of 19 BK channel subunits and interaction partners in brain samples. We will then assess how virally-mediated local silencing of the most promising genes affects ethanol self-administration. In a third Specific Aim, a similar functional approach will be used to probe the contribution of these genes to GABAergic neurotransmission in the amygdala and synaptic plasticity in the nucleus accumbens, in collaboration with INIA-West investigators. The proposed experiments are expected to uncover the contribution of BK channel interactome to ethanol self-administration and potentially pinpoint novel molecular targets for the treatment of alcoholism. PUBLIC HEALTH RELEVANCE: This research project investigates the contribution of a major potassium channel of the brain to excessive alcohol drinking. We anticipate this work to provide integrated insights into the molecular mechanisms mediating the behavioral and cellular effects of alcohol. Translational implications include the identification of novel targets for the development of a more efficient treatment of alcoholism.

描述（由申请人提供）：酒精滥用和依赖影响估计占美国人口的8.5％，并负责大量健康和社会成本。大型电导钙激活的钾（BK）通道以及已知与BK A亚基相互作用的20％以上的蛋白质已被INIA-WEST财团鉴定为乙醇偏好的潜在遗传决定因素。在本提案中，我们假设相关大脑区域中酒精对BK通道相互作用网络的扰动会导致过度饮酒。我们将把研究重点放在乙醇（Accumbens，杏仁核和前额叶皮层）的动机作用的神经记录上。我们进一步假设乙醇诱导的BK通道相互作用组的重组介导了在乙醇依赖性动物中观察到的神经传递和突触可塑性的变化。为了检验这些假设，我们将量化和操纵BK亚基的关键相互作用伙伴的表达水平。第一个具体目的是表征对BK通道的两个神经元辅助亚基中任何一个缺乏敲除小鼠的表型。我们将使用乙醇中毒，耐受性和戒断的测定，以及自愿饮酒的范式，导致中度或过度乙醇的摄入量。第二个具体目的是绘制伏伏核，杏仁核和前额叶皮层中已知的BK相互作用伙伴的表达，并评估过度乙醇暴露如何改变其蛋白质水平。将利用创新的蛋白质测定法，以同时定量19 BK通道亚基和脑样本中的相互作用伴侣。然后，我们将评估最有前途的基因的病毒介导的局部沉默如何影响乙醇自我给药。在第三个特定目的中，将使用类似的功能方法来探测这些基因对杏仁核中GABA能神经传递的贡献，并与Inia-West研究者合作。提出的实验预计将发现BK通道相互作用组对乙醇自我给药的贡献，并有可能确定酗酒治疗的新型分子靶标。公共卫生相关性：该研究项目调查了大脑主要钾通道对过量饮酒的贡献。我们预计这项工作可以为介导酒精的行为和细胞作用的分子机制提供综合见解。翻译的意义包括鉴定出更有效治疗酒精中毒的新目标。