ETHANOL EFFECTS ON GLYCINE RECEPTOR/CHANNEL FUNCTION

乙醇（EtOH）是一种有效的大脑刺激剂和添加剂药物。 新出现的证据表明，甘氨酸受体/通道（GlyR）是 对乙醇浓度敏感。 以来 甘氨酸抑制神经元活动，GlyR功能的增强将 预计将增强神经元抑制，并可能有助于 EtOH的神经毒性作用。 因此，我们建议研究 乙醇对甘氨酸诱发多巴胺能神经元反应的影响 从大脑的腹侧被盖区（VTA），奖励中心 因为吸毒 本研究的总体目标是调查 EtOH改变GlyR功能的机制有助于 酒精对中枢神经系统（CNS）的影响。 到 为了实现这一目标，将检验以下三个假设。 假设I是EtOH与GlyR相互作用。 EtOH调节 多巴胺能神经元的兴奋性通过改变GlyRs的功能。 假设II是EtOH与GlyR的相互作用受到以下因素的调节： GlyR的蛋白质磷酸化状态，细胞内GlyR的蛋白质磷酸化状态。 PKA、PKC和G蛋白的活性。假设3：GlyR 结构，多巴胺能神经元的细胞内C1浓度， 因此，甘氨酸诱导的反应及其对EtOH的反应 随着发展而变化。 这些假设将在腹侧被盖区神经元上进行检验 新鲜分离自新生和成熟大鼠。 全细胞贴片- 钳夹技术（尤其是短杆菌肽穿孔贴片技术）将 用于记录甘氨酸诱导的反应，包括膜电流， 潜在的和自发放电的改变， EtOH的存在。蛋白激酶的特异性激活剂和抑制剂 A和C和G蛋白将被用来确定酶的途径 参与乙醇对GlyR的任何影响。 这些研究将 显著推进我们对乙醇对CNS影响的理解 GlyR在分子和细胞水平上的作用。 更好地了解 EtOH在大脑中的作用将提高我们对相关疾病的理解。 加强机制，这将反过来促进 确定可能对治疗有价值的策略 酒精滥用和胎儿酒精综合症

Ethanol (EtOH) is an effective brain depressant and an additive drug. Emerging evidence suggests that glycine receptor/channels (GlyRs) are sensitive to pharmacologically relevant concentrations of EtOH. Since glycine inhibits neuronal activity, potentiation of GlyR function would be expected to enhance neuronal inhibition and perhaps contribute to the neuronal depressant effects of EtOH. Therefore, we propose to examine the effects of EtOH on glycine-induced responses of dopaminergic neurons from the ventral tegmental area (VTA) of the brain, the reward center for drug abuse. The overall objective of this study is to investigate the mechanisms by which EtOH alteration of GlyR function contributes to the central nervous system (CNS) consequences of alcohol in vivo. To achieve this objective the following three hypotheses will be tested. HYPOTHESIS I is that EtOH interacts with the GlyR. EtOH regulates the excitability of dopaminergic neurons by altering functions of GlyRs. HYPOTHESIS II is that EtOH interactions with the GlyR are modulated by the protein phosphorylation status of the GlyR, the intracellular activity of PKA, PKC and G-proteins. HYPOTHESIS III is that GlyR structure, intracellular C1-concentration of dopaminergic neurons and, consequently, glycine-induced responses and their response to EtOH change with development. These hypotheses will be tested on VTA neurons freshly isolated from both neonatal and mature rats. Whole-cell patch- clamp technique (especially gramicidin perforated patch technique) will be used to record glycine-induced responses, including membrane current, potential and the alteration of spontaneous firing in the absence and presence of EtOH. Specific activators and inhibitors of protein kinases A and C and of G-proteins will be used to identify the enzyme pathways involved in any effects, of EtOH on GlyRs. These studies will significantly advance our understanding of the effects of EtOH on CNS GlyRs at the molecular and cellular levels. A better knowledge of the actions of EtOH in the brain will improve our understanding of related reinforcement mechanisms, which will, in turn, facilitate the identification of strategies which might be of value in the treatment of alcohol abuse and fetal alcohol syndrome.