NEUROBIOLOGY OF ETHANOL/GABA INTERACTION

乙醇/GABA 相互作用的神经生物学

• 批准号: 2046152
• 负责人: Hermes H Yeh
• 金额: $ 8.78万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-03-01 至 1997-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2046152
• 关键词: antisense nucleic acid; electrophysiology; ethanol; gamma aminobutyrate; laboratory rat; membrane channels; molecular biology; neurobiology; neurotransmitter receptor; phosphorylation; polymerase chain reaction; retinal bipolar neuron; retinal ganglion; toxicant interaction; voltage /patch clamp; antisense nucleic acid; electrophysiology; ethanol; gamma aminobutyrate; laboratory rat; membrane channels; molecular biology; neurobiology; neurotransmitter receptor; phosphorylation; polymerase chain reaction; retinal bipolar neuron; retinal ganglion; toxicant interaction; voltage /patch clamp

Despite well-known psychoactive effects, the mechanism of action of ethanol in the central nervous system (CNS) is not well understood. Rather than disrupting the integrity of the lipid bilayer of neuronal membranes in a non-specific way, it is becoming increasingly clear that acute exposure to clinical concentrations of ethanol can affect the function of a number of neurotransmitter receptors, such as those for gamma- aminobutyric acid (GABA), glutamate and serotonin. The overall goal of this research project is to contribute to an understanding of the mechanisms by which acute exposure to ethanol modulates the function of ligand-activated channels, specifically, on the modulation of the GABA-A receptor by ethanol. The experiments combine patch clamp electrophysiological and molecular biological approaches and will be conducted using bipolar cells and ganglion cells of the rat retina. Specific Aim l proposes to test the hypothesis that the GABA-A receptor is a direct target site for the modulatory effect of ethanol and that phosphorylation mediated by intracellular intermediaries can act secondarily to prime the sensitivity of the receptor to ethanol. We will analyze ethanol-induced changes in stationary and non-stationary channel kinetics of the GABA-A receptor and determine whether intracellular dialysis of agents which promote or prevent phosphorylation can influence the outcome of a synergistic interaction between ethanol and GABA. Specific Aim 2 will test the hypothesis that the subunit composition of the GABA-A receptor is critical in conferring sensitivity to GABA potentiation by ethanol such that neurons whose responses to GABA are potentiated by ethanol can be grouped into predictable patterns of subunit profiles. We will determine by "expression profiling" the GABA-A receptor subunit composition of those bipolar cells and ganglion cells which have been examined electrophysiologically in Specific Aim 1, using a combination of molecular biological techniques involving antisense RNA (aRNA) amplification and polymerase chain reaction (PCR). In this way, a composite of GABA-A receptor subunits can be revealed from each neuron and this can be correlated with functional data acquired from the same neuron. Overall, this project will contribute to resolving some of the most pressing issues related to ethanol action on GABA inhibitory mechanisms as well as to determining the molecular basis of functional heterogeneity in ethanol-GABA interactions in the mammalian CNS.

尽管众所周知的精神活性作用，但 中枢神经系统（CNS）中的乙醇尚不清楚。相当 而不是破坏神经元膜的脂质双层的完整性 以非特异性的方式，越来越明显 暴露于乙醇的临床浓度会影响 许多神经递质受体，例如γ- 氨基丁酸（GABA），谷氨酸和5-羟色胺。 总体目标 该研究项目是为了理解 急性暴露于乙醇的机制调节功能 配体激活的通道，特别是在GABA-A的调节中 乙醇的受体。实验结合了斑块夹 电生理和分子生物学方法，将是 使用大鼠视网膜的双极细胞和神经节细胞进行。 具体目的L提出测试GABA-A受体是的假设 乙醇调节作用的直接目标位点， 细胞内中介介导的磷酸化可以作用 其次是为了使受体对乙醇的敏感性增强。我们将 分析乙醇诱导的固定通道和非平稳通道的变化 GABA-A受体的动力学，并确定细胞内是否 促进或预防磷酸化的药物的透析可能会影响 乙醇与GABA之间的协同相互作用的结果。 具体目标2将检验以下假设。 GABA-A受体对于赋予对GABA的敏感性至关重要 乙醇的增强，使得对GABA反应的神经元是 可以将乙醇增强，可以分为可预测的亚基模式 概况。 我们将通过“表达谱分析” GABA-A受体来确定 那些具有双极细胞和神经节细胞的亚基组成 使用A 涉及反义RNA的分子生物学技术的组合 （ARNA）扩增和聚合酶链反应（PCR）。这样， 可以从每个神经元中揭示GABA-A受体亚基的复合物 这可以与从同一神经元中获取的功能数据相关。 总体而言，该项目将有助于解决一些最大的 压迫与乙醇对GABA抑制性机制有关的问题 以及确定功能异质性的分子基础 哺乳动物中枢神经系统中的乙醇-GABA相互作用。