Dissecting the Role of Nucleus Basalis Magnocellularis Circuitry in Basolateral Amygdala Physiology and Regulation of Anxiety Following Chronic Ethanol Exposure and Withdrawal

项目总结 戒酒期间的焦虑会导致高复发率，并仍然是酒精使用的障碍 障碍(AUD)治疗。基底外侧杏仁核(BLA)是一个参与脑部疾病进展的区域。 酒精依赖会促进寻求奖励和焦虑等情绪行为。我们的实验室有 研究表明，谷氨酸/GABA神经传递的特定输入改变发生在慢性 乙醇/戒断。例如，终纹(ST)谷氨酸能传入显示突触前易化。 在戒烟期间。外侧针旁细胞和BLA GABA能系统的局部中间神经元 慢性乙醇的不同结果：LPCS显示GABA释放减少，而局部 中间神经元无明显变化。总之，GABA能/谷氨酸能传入的这些变化 在酒精戒断过程中，BLA锥体神经元处于过度兴奋状态。然而，目前仍不清楚 乙醇如何改变上游调节系统的神经生理学，或者这些投射 影响焦虑类行为的产生。BLA接受密集的胆碱能投射。 大细胞基底核对神经元兴奋性和神经递质释放的调节作用 作用于GABA能中间神经元、谷氨酸能终末和BLA锥体神经元。这些不同的 神经元隔室含有烟碱型和毒鼠型乙酰胆碱受体的不同组合 (n/mAChRs)，它们共同调节厌恶记忆的形成。慢性病的影响 乙醇对大鼠黑质突触生理的影响及其在调节戒断焦虑样行为中的作用 BLA尚不清楚。我们的初步数据引出了一个中心假设，即慢性乙醇调节失调 胆碱能调节BLA的传入和增强戒断的神经生理症状。我们 我将通过两个具体的目标来解决这一假设。在目标1中，我们将主要使用光生和 兴奋性损伤研究操纵NBM终末活性和测定GABA/谷氨酸 神经生理学与全细胞膜片钳电生理学。我们假设慢性病 乙醇/戒断上调NBM胆碱能输入并增强谷氨酸和GABA能 突触前可塑性。在目标2中，我们将使用化学遗传学和损伤实验来操纵 胆碱能输入，并使用高架零迷宫、明/暗盒、 和露地测试。我们假设，胆碱能环路的中断将改善 焦虑样的行为。了解NBM-BLA电路可能会导致更多潜在的新目标 临床环境下酒精戒断所致焦虑的有效治疗。

PROJECT SUMMARY Anxiety during alcohol withdrawal contributes to high relapse rates and remains an obstacle for alcohol use disorder (AUD) treatment. The basolateral amygdala (BLA) is a brain region involved in the progression of alcohol dependence that facilitates reward-seeking and emotional behaviors like anxiety. Our laboratory has shown that input-specific alterations in glutamate/GABA neurotransmission occur following chronic ethanol/withdrawal. For example, stria terminalis (ST) glutamatergic afferents exhibit presynaptic facilitation during withdrawal. Lateral paracupsular cells (LPCs) and local interneurons of the BLA GABAergic system express different outcomes to chronic ethanol: LPCs show an attenuation of GABA release, while local interneurons show no significant change. Together, these alterations in GABAergic/glutamatergic afferents yield a hyperexcitable state in BLA pyramidal neurons during alcohol withdrawal. However, it remains unclear how ethanol alters the neurophysiology of upstream modulatory systems, or whether these projections influence the generation of anxiety-like behaviors. The BLA receives dense cholinergic projections from the nucleus basalis magnocellularis (NBM) to modulate neuronal excitability and neurotransmitt er release by acting on GABAergic interneurons, glutamatergic terminals, and BLA pyramidal neurons. These distinct neuronal compartments contain different combinations of nicotinic and muscarinic acetylcholine receptors (n/mAChRs) that collectively function to regulate the formation of aversive memories. The effects of chronic ethanol on NBM synaptic physiology and their roles in mediating withdrawal-induced anxiety-like behavior in the BLA is unknown. Our preliminary data leads to the central hypothesis that chronic ethanol dysregulates cholinergic modulation of BLA afferents and potentiates the neurophysiological symptoms of withdrawal. We will address this hypothesis through two Specific Aims. In Aim 1, we will primarily employ optogenetic and excitotoxic lesion studies to manipulate the activity of NBM terminals and measure GABA/glutamate neurophysiology with whole-cell patch clamp electrophysiology. We hypothesize that chronic ethanol/withdrawal upregulates NBM cholinergic input and enhances glutamatergic and GABAergic presynaptic plasticity. In Aim 2, we will use chemogenetics and lesion experiments to manipulate the cholinergic inputs and measure anxiety-like behavior using assays like the elevated zero maze, light/dark box, and open field test. We hypothesize that disruption of the cholinergic circuit will ameliorate the progression of anxiety-like behaviors. Understanding the NBM-BLA circuit might lead to potential novel targets for more effective treatments of alcohol withdrawal-induced anxiety in the clinical setting.