ALCOHOL EFFECTS ON RECEPTOR G PROTEIN COUPLING

拟议的研究将检查慢性乙醇对G- 蛋白偶联受体功能。脑内神经递质受体 特定的大脑区域被认为会受到乙醇的影响。G- 蛋白偶联受体构成了一类主要的受体 脑，并检测到G蛋白水平的变化 乙醇管理。然而，关于这一问题的数据很少 慢性酒精对受体偶联G蛋白活性的影响这个 拟议的研究将确定在功能上的变化 G蛋白偶联受体在特定脑区的活性为 受长期酒精饮食的影响。初步研究将涉及 5-HT1a和阿片受体活性的解剖学观察 接受酒精的大鼠的大脑和对照组的大鼠的大脑。此外, 小脑中几种G蛋白偶联受体的活性 将会被检查。这些实验将使用一种新技术， 直接测量受体介导的G蛋白激活 解剖学和药理学上的特殊方式：体外 激动剂刺激的[35S]GTP-GammaS结合的放射自显影。区域 其中受体-G蛋白功能被乙醇改变 然后将在[35S]GTPGammaS膜结合中检查给药 化验。膜[35S]GTP-GammaS结合分析将允许 激动剂效力或最大作用变化的测定 在注射乙醇后。基础和激动剂的饱和度分析- 受刺激的[35S]GTP-GammaS结合将用于确定是否 受体对G蛋白的催化激活发生了变化， 或者在受体偶联的G蛋白对GTP伽马S的亲和力上， 在长期服用乙醇后。这些研究将结合在一起 使用放射性标记的受体拮抗剂进行受体结合分析。 受体结合的饱和分析将被执行以测量任何 慢性阻塞性肺疾病影响系统中受体数量的变化 乙醇管理。此R03应用程序的目的是 提供将用于在 未来，通过允许产生关于影响的假设 乙醇对G蛋白偶联受体功能活性的影响。

The proposed studies will examine the effects of chronic ethanol on G- protein-coupled receptor function. Neurotransmitter receptors in specific brain regions are thought to be affected by ethanol. The G- protein-coupled receptors constitute a major class of receptors in brain, and changes in G-protein levels have been detected after chronic ethanol administration. However, there is little data regarding the effects of chronic ethanol on receptor-coupled G-protein activity. The proposed studies will determine whether changes in the functional activity of G-protein-coupled receptors in specific brain regions are affected by a chronic ethanol diet. Initial studies will involve an anatomical survey of the activity of 5-HT1A and opioid receptors in brains from rats that receive ethanol and control rats. In addition, the activity of several G-protein-coupled receptors in the cerebellum will be examined. These experiments will use a novel technique that directly measures receptor-mediated G-protein activation in an anatomically and pharmacologically specific manner: in vitro autoradiography of agonist-stimulated [35S]GTPgammaS binding. Regions in which receptor-G-protein function is altered by ethanol administration will then be examined in [35S]GTPgammaS membrane binding assays. Membrane [35S]GTPgammaS binding assays will allow the determination of changes in the potency or maximal effect of agonists after ethanol administration. Saturation analysis of basal and agonist- stimulated [35S]GTPgammaS binding will be used to determine whether changes occur in the catalytic activation of G-proteins by receptors, or in the affinity of the receptor-coupled G-proteins for GTPgammaS, after chronic ethanol administration. These studies will be combined with receptor binding assays using radiolabeled receptor antagonists. Saturation analysis of receptor binding will be performed to measure any changes in receptor number in those systems that are affected by chronic ethanol administration. The purpose of this R03 application is to provide data that will be used to develop an R01 application in the future by allowing the generation of hypotheses regarding the effects of ethanol on G-protein-coupled receptor functional activity.