Alcohol Actions-Molecular Targets on Brain Proteins

描述(申请人提供)：乙醇的急性作用是通过结合蛋白质上的特定位点来实现的。我们建议结合分子、电生理和行为学的方法来阐明乙醇对GABAA和甘氨酸受体的作用，这是乙醇的两个关键分子靶点。目的1阐明在异构体GABAA受体中形成乙醇结合部位的氨基酸的具体位置和方向。我们正在进行的甘氨酸受体研究表明，甘氨酸受体的生理调节剂锌在乙醇对这些受体的作用中是重要的，目标1也将定义这种相互作用的分子基础。在目标2中，我们将使用GABAA和甘氨酸受体敲除和敲除小鼠来明确乙醇对这些受体的作用所产生的行为影响。目的3将评估受体突变对神经元的影响。这将通过对关键大脑区域的基因组分析和对中脑边缘奖赏通路的电生理学研究来实现。目的3还将确定慢性酒精摄入对腹侧被盖区和伏隔核基因表达变化的影响，这两个区域是中脑边缘通路的两个关键区域。这些研究将使用野生型小鼠和带有GABAA或甘氨酸受体的小鼠，这些受体被设计为对乙醇作用不敏感。这一新的方法将使我们能够将基因表达(和行为，目标2)的变化与酒精对特定受体的作用联系起来。这项工作的长期目标是确定关键的蛋白质位点，这些蛋白质位点可以作为新疗法的靶点，以减轻酒精强化、依赖和复发。与公共健康相关尽管酒精(乙醇)已经被消费了数千年，但我们对它对大脑产生影响的方式知之甚少。一个重要的进展是确定参与神经元之间通信的特定蛋白质(神经递质受体和离子通道)作为乙醇的靶标。我们将使用不同的技术，从分子水平到行为水平，使用小鼠的突变和其他新技术来定义乙醇如何作用于这些蛋白质，最终目标是确定关键的蛋白质位点，这些蛋白质可以作为缓解酒精成瘾的新疗法的靶点。

DESCRIPTION (provided by applicant): The acute effects of ethanol are mediated by binding to specific sites on proteins. We propose to elucidate the actions of ethanol on the GABAa and glycine receptors, two critical molecular targets for ethanol, by combining molecular, electrophysiological and behavioral methods. Aim 1 will elucidate the specific location and orientation of the amino acids that form the ethanol binding site in heteromeric GABAa receptors. Our ongoing studies of glycine receptors suggest that zinc, a physiological modulator of glycine receptors, is important for ethanol action on these receptors, and Aim 1 will also define the molecular basis of this interaction. In Aim 2, we will use GABAa and glycine receptor knockout and knockin mice to specify the behavioral effects of ethanol that are due to actions on these receptors. Aim 3 will evaluate the neuronal consequences of receptor mutation. This will be accomplished by genomic analysis of key brain regions and by electrophysiological study of the mesolimbic reward pathway. Aim 3 will also define effects of chronic ethanol consumption on global gene expression changes in the ventral tegmental area and nucleus accumbens, the two key areas of the mesolimbic pathway. These studies will use wild type mice and mice with GABAa or glycine receptors that are engineered to be insensitive to ethanol action. This novel approach will allow us to link changes in gene expression (and behavior, Aim 2) to alcohol actions on specific receptors. The long-range goal of this work is to define key protein sites that can serve as targets for new therapies to alleviate alcohol reinforcement, dependence an relapse. PUBLIC HEALTH RELEVANCE Even though alcohol (ethanol) has been consumed for thousands of years, we know remarkably little about the way it produces its effects on the brain. An important advance was the identification of specific proteins (neurotransmitter receptors and ion channels) involved in communication between neurons as a target for ethanol. We will define how ethanol acts on these proteins using different techniques, ranging from the molecular to the behavioral level, using mutations in mice and other new technologies, with the final objective of defining key protein sites that can serve as targets for new therapies to alleviate alcohol addiction.