Role of BK Channel Interactome in Excessive Ethanol Drinking

描述（由申请人提供）：酒精滥用和依赖影响估计8.5%的美国人口，并负责大量的健康和社会成本。大电导钙激活钾（BK）通道，连同超过20%的蛋白质已知与BK α亚基相互作用，已被INIA-West协会确定为乙醇偏好的潜在遗传决定因素。 在目前的建议中，我们假设，BK通道相互作用网络的干扰，酒精在相关的大脑区域有助于过量饮酒。我们将集中我们的调查对神经回路subtending乙醇的动机影响（核杏仁核和前额叶皮层）。我们进一步假设，乙醇诱导的重组BK通道相互作用体介导的变化，在乙醇依赖性动物中观察到的神经传递和突触可塑性。 为了检验这些假设，我们将量化和操纵BK α亚基的关键相互作用伙伴的表达水平。第一个具体目标是表征BK通道的两个神经元辅助亚基中的任一个缺陷的敲除小鼠的表型。我们将使用酒精中毒，耐受性和戒断试验，以及自愿饮酒导致适度或过量酒精摄入的范例。 第二个具体目标是绘制已知BK相互作用伙伴在丘脑核、杏仁核和前额叶皮层中的表达，并评估过量乙醇暴露如何改变其蛋白质水平。一种创新的蛋白质测定将被利用的同时定量的19 BK通道亚基和相互作用的合作伙伴在大脑样本。然后，我们将评估病毒介导的最有前途的基因的局部沉默如何影响乙醇自我管理。在第三个具体目标中，将使用类似的功能方法来探测这些基因对杏仁核中GABA能神经传递和突触可塑性的贡献，与INIA-West研究人员合作。 拟议的实验有望揭示BK通道相互作用组对乙醇自我给药的贡献，并可能确定治疗酒精中毒的新分子靶点。 公共卫生相关性：该研究项目调查了大脑的主要钾通道对过量饮酒的贡献。我们预计这项工作提供了综合的见解的分子机制介导的行为和细胞的酒精的影响。翻译的影响包括识别新的目标，为发展一个更有效的治疗酗酒。

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.