SYNAPTIC GABA AND GLUTAMATE FUNCTION IN EXCESSIVE ETHANOL SELF-ADMINISTRATION

酒精引起的睡眠紊乱在酒精中毒的进展中起着关键作用，并预示着复发。 对于一些失眠症患者来说，酒精的镇静作用是睡前饮酒和最终滥用酒精的途径。持续滥用乙醇会导致睡眠回路的长期变化，这种变化会持续到停止使用乙醇之后。该项目的总体目标是确定睡眠回路对酒精的细胞适应的关键分子事件，最终目标是确定用于治疗乙醇破坏正常睡眠的新型药物靶点。丘脑是睡眠/觉醒周期和睡眠期间发生的大脑节律的主要发生器。对这些节律最好的理解是与第二阶段睡眠相关的丘脑纺锤体振荡，这种振荡会响应于 急性酒精中毒在酗酒者中，纺锤波减少，取而代之的是不太平静的随机眼动（REM）睡眠。因此，了解纺锤波睡眠的命运是一个与乙醇的强化效应直接相关的重要问题，因为一些酗酒者为了改善睡眠质量而重新饮酒。没有实验室在灵长类动物模型或具有良好特征的饮酒时间表的动物中解决这些变化的机制。使纺锤波流经大脑的引擎是一种低阈值钙电流，由三种已知类型的T型钙通道介导。我们最近发现，在丘脑中继细胞中发现的T通道转录本对乙醇非常敏感，在生理上有意义的乙醇浓度（10- 17 mM）的低端显示出急性增强。令人惊讶的是，这种电流似乎被浓度远高于这个范围的乙醇抑制。在本项目的目标1中，我们将通过在两种制剂（猴子和大鼠）中进行全细胞斑片记录，在过量饮酒的条件下和在接近戒断峰值的条件下，确定慢性饮酒动物的背外侧膝状体核中T通道是否功能受损。在目标2中，我们将确定与补丁记录是否T通道功能降低，在丘脑网状核的慢性饮酒动物。目的3的实验将提供一个补充检查的T型通道的分子表达模式，在猴子和大鼠使用定量RT-PCR技术。

Sleep perturbations by ethanol play a key role in the progression of alcoholism, and are predictive of relapse. For some insomniacs, the sedative effects of ethanol are the pathway to bedtime alcohol consumption and eventual abuse. Continued abuse of ethanol leads to long-term changes in sleep circuitry that last well beyond the cessation of ethanol administration. The overall goal of this project is to determine the key molecular events that underlie cellular adaptation of sleep circuitry to alcohol, with the eventual goal of identifying novel drug targets for treatment of ethanol's disruption of normal sleep. The thalamus is a primary generator of sleep/wake cycles and the brain rhythms that occur during sleep. The best understood of these rhythms is the thalamic spindle oscillation associated with Stage II sleep, which is enhanced in response to acute alcohol administration. In alcoholics, spindle waves are diminished and are replaced with less-restful random eye movement (REM) sleep. Understanding the fate of spindle wave sleep is thus a vital question that directly relates to the reinforcement effects of ethanol, since some alcoholics return to drinking in an effort to improve the quality of their sleep. No laboratory has addressed the mechanism of these changes in a primate model, or in animals with well-characterized drinking schedules. The engine that allows spindle waves to flow through the brain is a low threshold calcium current, mediated by T-type calcium channels that come in three known varieties. We have recently shown that the T-channel transcript found in thalamic relay cells is exquisitely sensitive to ethanol, showing acute enhancement at the low end of physiologically meaningful ethanol concentrations (10-17mM). Amazingly, this current appears to be inhibited by concentrations of ethanol much above this range. In Aim 1 of this project we will determine whether the T channel is functionally impaired in the dorsal lateral geniculate nucleus of chronic drinking animals by performing whole cell patch recordings in two preparations, monkeys and rats, in a condition of excessive drinking and in a condition near the peak of withdrawal. In Aim 2, we will determine with patch recordings whether the T channel function is reduced in the thalamic reticular nucleus of chronic drinking animals. The experiments of Aim 3 will provide a complementary examination of the molecular expression patterns of T-type channels in monkeys and rats using quantitative RT-PCR techniques.