CELLULAR MECHANISM OF ACTION OF ALCOHOL

酒精的细胞作用机制

• 批准号: 2457459
• 负责人: TOSHIO NARAHASHI
• 金额: $ 14.57万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-07-01 至 1999-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2457459
• 关键词: GABA receptor; acetylcholine; active sites; age difference; chloride channels; cyclic AMP; dorsal root; electrophysiology; ethanol; excitatory aminoacid; hippocampus; human fetus tissue; kidney cell; laboratory rat; membrane channels; neurons; neurotoxins; phosphorylation; protein kinase; serotonin; spinal ganglion; temperature; tissue /cell culture; transfection; voltage /patch clamp

The long-term objectives of the proposed study are to elucidate the mechanisms by which alcohols exert their toxic actions on the nervous system. Specific attention is focused on the mechanisms underlying the interactions of ethanol and longer-chain alcohols with ion channels by using patch clamp techniques as applied to mammalian neurons. Our previous studies as well as those by other investigators have laid out the groundwork along this line, establishing the phenomenological aspects of alcohol interactions with certain types of ion channels including those activated by GABA, excitatory amino acid (EAA), acetylcholine (ACh) and 5-hydroxytryptamine (5-HT), and voltage-activated sodium, potassium and calcium channels. However, conflicting data have been obtained for certain types of ion channels, and in many cases no detailed mechanisms of alcohol action have been elucidated. The proposed study is aimed at solving some of these problems. Thus the proposed projects will focus on elucidation of the mechanisms of ethanol and longer-chain alcohols on the GABA/A receptor-channel complex. Both whole-cell and single-channel patch clamp techniques will be applied to rat dorsal root gang lion, hippocampal and cortical neurons, and cerebellar Purkinje and granular layer neurons. The specific aims will be concerned with the modulation of open and/or closed channels, dependence of alcohol action on neuron type, animal age and experimental temperature, the role of intracellular components in alcohol action, and the comparison of subtypes and subunits of receptor-channel complex. For the GABA receptor subunit study, human embryonic kidney (HEK-293) cells, in which various combinations of GABA subunits have been transfected, will be used to determine the role of each subunit in alcohol action. Attention will be focused not only on changes in tee amplitude of GABA-induced currents, but also on changes in the rate of desensitization of the currents. The results of the proposed study will provide the basis for the mechanisms of alcoholism and for approaches to prevention and cure of the disorder.

拟议研究的长期目标是阐明 酒精对神经的有毒作用的机制 系统。具体关注的重点是该机制 乙醇和长链醇与离子通道的相互作用 使用贴片夹技术应用于哺乳动物神经元。我们的 先前的研究以及其他研究人员的研究 沿着这条线的基础，建立了现象学方面 与某些类型的离子通道的酒精相互作用 那些被GABA，兴奋性氨基酸（EAA），乙酰胆碱（ACH）激活的人 和5-羟化胺（5-HT）和电压激活的钠，钾 和钙通道。但是，已经获得了相互矛盾的数据 某些类型的离子通道，在许多情况下没有详细的机制 酒精作用已被阐明。拟议的研究针对 解决其中一些问题。因此，拟议的项目将重点放在 阐明乙醇和长链酒精的机制 GABA/A受体通道复合物。全电池和单通道补丁 夹具技术将应用于大鼠背根帮的狮子， 海马和皮质神经元，小脑Purkinje和颗粒状 层神经元。具体目标将与调制有关 开放和/或封闭的通道，酒精作用对神经元的依赖性 类型，动物年龄和实验温度，细胞内的作用 酒精作用中的组成部分以及亚型和亚基的比较 受体通道复合物。对于GABA受体亚基研究，人类 胚胎肾（HEK-293）细胞，其中各种GABA组合 亚基已被转染，将用于确定 酒精作用中的每个亚基。注意不仅会集中在 GABA诱导的电流的TEE振幅变化，但也会变化 在电流的脱敏速率上。结果 拟议的研究将为酒精中毒的机制提供基础 以及预防和治愈该疾病的方法。