Novel Mechanisms of Ethanol Potentiation of Rat Basolateral Amygdala GABA

描述（由申请人提供）：尽管酒精（EtOH）是已知最古老的滥用药物之一，但EtOH作用的神经机制尚不完全清楚，因此，很难理解一个人是如何失去对正常EtOH饮酒的控制而导致酗酒的。酒精中毒通常与焦虑和压力障碍共病，虽然这种共病的病因尚不完全清楚，但有人提出，由于EtOH长期以来一直被认为是一种抗焦虑剂，一些人可能会将其作为一种自我治疗焦虑或压力状态的方式。从高水平的EtOH中退出也被证明会导致焦虑/压力状态的增强，这可能会导致持续的EtOH使用和滥用。因此，假设EtOH可能主要影响介导应激反应和焦虑的中枢神经系统（CMS）区域，如基底外侧杏仁核（BLA）。脑左脑a中的gaba能突触在焦虑等情绪反应中起着重要作用。广泛的研究表明，至少部分EtOH对CMS的影响是由于通过突触前和突触后位点在许多脑区域增加gaba能神经传递。去甲肾上腺素（NE）与压力和焦虑症密切相关，已被证明可以增加BLA中GABA的释放。我们的初步数据表明，NE受体拮抗剂可以阻断EtOH对gaba能中间神经元子集的增强作用。因此，我们假设乙炔乙酸乙酯在一定程度上通过ne -受体活性增强了gaba能在BLA中的传递。第一个具体目标是确定急性EtOH在BLA中两个中间神经元亚群中增强gaba能神经传递的机制。这一目标将依靠全细胞膜片钳电生理方法来确定EtOH对BLA组织切片的影响是突触前和/或突触后，利用主要关注诱发抑制性突触后电流（IPSCs）自发和微型IPSCs的方案组合和配对脉冲促进研究来分离EtOH增强gaba能神经传递的突触前和突触后成分。第二个具体目标是确定NE在BLA gaba能突触与EtOH相互作用的机制，以及哪种NE受体亚型介导这种作用。我们将使用与上述类似的方法以及NE受体激动剂和拮抗剂的组合来分离该脑区域中NE- etoh相互作用的机制。第三个具体目标将侧重于通过吸入室暴露慢性间歇性EtOH暴露对随后急性EtOH暴露的影响以及目标1和目标2研究的NE-EtOH相互作用。

DESCRIPTION (provided by applicant): Although alcohol (EtOH) is one of the oldest known drugs of abuse, the neuroliogical mechanisms underlying EtOH's actions are not fully known and as such, it remains difficult to understand how one may lose control of normal EtOH drinking resulting in alcoholism. Alcoholism is often found to be comorbid with anxiety and stress disorders, and while the eitology of this comorbidity is not fully known it has been proposed that since EtOH has long been known as an anxiolytic agent, some may imbibe as a way to selfmedicate their anxious or stressed states. Withdrawal from high EtOH levels has also been shown to result in an enhancement of anxious/stressed states, which may result in continued EtOH use and abuse. Therefore, it is hypothesized that EtOH may predominantly effect areas of the central nervous system (CMS) which mediate stress responses and anxiety, such as the basolateral amygdala (BLA). GABAergic synapses in the BLA have been shown to play a major role in emotional responses such as anxiety. Extensive research has shown that at least part of EtOH's effects in the CMS are due to an increase in GABAergic neurotransmission in numerous brain regions through both pre- and post-synaptic sites. Norepinephrine (NE), which is intimately linked to stress and anxiety disorders, has been shown increase GABA release in the BLA. Our preliminary data shows that NE receptor antagonists can block the potentiating effect of EtOH at a subset of GABAergic interneurons. Therefore it is hypothesized that EtOH enhances GABAergic transmission in the BLA in part via NE-receptor activity. The first specific aim is to determine the mechanisms by which acute EtOH enhances GABAergic neurotransmission at two subset of interneurons in the BLA. This aim will rely on whole-cell patch clamp electrophysiological methods to determine if the effects of bath application of EtOH on slices of BLA tissue are pre- and/or post-synaptic utilizing a combination of protocols focusing mainly on evoked inhibitory postsynaptic currents (IPSCs) spontaneous and miniature IPSCs and paired-pulses facilitation studies to isolate pre- and post-synaptic components of the EtOH enhancement of GABAergic neurotransmission. The second specific aim will determine the mechanism by which NE interacts with EtOH at BLA GABAergic synapse and which NE-receptor subtype mediates this effect. We will be using similar methods as described above as well as combinations of NE receptor agonists and antagonists to isolate the mechanism of the proposed NE-EtOH interaction in this brain region. The third specific aim will focus on the effects of chonic intermittent EtOH exposure via inhalation chamber exposure on subsequent acute EtOH exposure and the NE-EtOH interaction studied aims 1 and 2.