Neuron-astrocyte interactions mediating ethanol and noradrenergic modulation of wake-promoting vPAG dopamine neurons

神经元-星形胶质细胞相互作用介导促醒 vPAG 多巴胺神经元的乙醇和去甲肾上腺素能调节

• 批准号: 10463298
• 负责人: Nicholas Petersen
• 金额: $ 3.2万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10463298
• 关键词: Acute; Adenosine; Adenosine Triphosphate; Adrenergic Agonists; Adrenergic Receptor; Agonist; Alcohol consumption; Alcohol withdrawal syndrome; American; Arousal; Astrocytes; Behavioral; Brain; Brain region; Calcium; Calcium Signaling; Cells; Chronic; Development; Dose; Electrophysiology (science); Ethanol; Fiber; Fluorescence Microscopy; G alpha q Protein; Goals; Human; Individual; Investigation; Measures; Mediating; Mental disorders; Midbrain structure; Modeling; Mus; Neuroglia; Neurons; Norepinephrine; Pathologic; Periodicity; Pharmacology; Phenylephrine; Photometry; Physiology; Population; Purines; Purinoceptor; Receptor Activation; Relapse; Reporting; Role; Series; Signal Transduction; Signaling Protein; Sleep; Sleep Disorders; Sleep disturbances; Sleeplessness; Slice; Stress; Synapses; Synaptic Transmission; Synaptic plasticity; System; Testing; Time; Viral; Wakefulness; Withdrawal; Work; alcohol abstinence; alcohol exposure; alcohol use disorder; alertness; alpha-1 adrenergic receptors; antagonist; chronic alcohol ingestion; comorbidity; disorder later incidence prevention; dopaminergic neuron; excitatory neuron; experience; experimental study; extracellular; fluorescence imaging; genetic approach; in vivo; locus ceruleus structure; midbrain central gray substance; mouse model; new therapeutic target; noradrenergic; novel; novel therapeutics; patch clamp; prevent; promoter; receptor expression; relapse risk; response; sensor; sleep difficulty; sleep regulation; therapeutic target; transmission process; virus genetics

PROJECT SUMMARY An estimated 50-70 million Americans suffer from a sleep disorder, which is commonly comorbid with many psychiatric illnesses, such as alcohol use disorder (AUD). Individuals with AUD frequently report insomnia, and sleep difficulty significantly increases the likelihood of relapse. Despite the overwhelming need to control sleep disturbances to aid in the prevention of relapse, the underlying physiology of wakefulness, particularly in the context of acute and chronic alcohol use, is not well understood. The brain’s noradrenergic system mediates many behavioral states, such as arousal, alertness, and stress. Acute and chronic alcohol exposure differentially alter the activity of noradrenergic neurons as well as the release, synthesis, and turnover of norepinephrine (NE) in the brain. Recently, our lab alongside collaborators delineated a novel arousal circuit from the noradrenergic locus coeruleus (LC) to dopaminergic neurons in the ventral periaqueductal gray (vPAGDA), where stimulation of this circuit promotes wakefulness. Acute ethanol exposure also increases excitatory drive and the activity of wake-promoting vPAGDA neurons; however, the mechanism is unknown. Specific activation of alpha1-adrenergic receptors (α1ARs) increases excitatory drive onto vPAGDA neurons and subsequently causes arousal. Interestingly, α1AR expression is particularly enriched on vPAG astrocytes, and activation of astrocytic Gq signaling is sufficient to promote wakefulness. While astrocytes are essential in mediating synaptic transmission in other brain regions through purinergic signaling, the role of neuron- astrocyte interactions in the vPAG during ethanol exposure requires further investigation. This F30 aims to test the central hypothesis that purinergic transmission from neighboring astrocytes mediates the noradrenergic modulation of vPAGDA neurons, and that ethanol potentiates this signaling. The goal is to elucidate the mechanistic details mediating the increase in excitatory drive onto the wake-promoting vPAGDA neurons by activation of α1ARs and the pathologic changes to the noradrenergic system following acute and chronic alcohol exposure. In Specific Aim 1 I will utilize pharmacological and viral genetic approaches combined with ex vivo fluorescence imaging and whole cell patch-clamp electrophysiology to examine the neuron-astrocyte interactions underlying noradrenergic modulation of vPAGDA neurons. In Specific Aim 2 I will use similar approaches in addition to a mouse model of chronic intermittent ethanol exposure to determine the changes in synaptic transmission onto vPAGDA neurons following acute and chronic ethanol exposure. The results of these studies will allow our lab, our collaborators, and others to further investigate the role of neuron- astrocyte physiology related to sleep dysfunction in alcohol use disorder. This will further the work towards identifying possible therapeutic targets for the development of novel therapies to alleviate disordered sleeping and aid in relapse prevention.

项目摘要 估计有500-70万美国人患有睡眠障碍，这通常与许多人合并 精神疾病，例如酒精使用障碍（AUD）。具有AUD的人经常报告失眠， 睡眠难以显着增加退休的可能性。尽管需要控制睡眠 有助于预防救济的干扰，觉醒的潜在生理学，尤其是在 大脑的北肾上腺素系统介导 许多行为状态，例如唤醒，机敏和压力。急性和慢性酒精暴露 差异改变了甲肾上腺神经元的活性以及释放，合成和周转率 大脑中的去甲肾上腺素（NE）。最近，我们的实验室与合作者一起描述了一个新颖的唤醒巡回赛 从去甲肾上腺素基因座（LC）到腹侧灰色的多巴胺能神经元 （vpagda），其中该电路的刺激会促进清醒。急性乙醇暴露也增加 兴奋性驱动和启动唤醒VPAGDA神经元的活性；但是，该机制尚不清楚。 α1-肾上腺素受体（α1AR）的特异性激活增加了兴奋性驱动到VPAGDA神经元和 有趣的是，α1AR表达在VPAG星形胶质细胞上特别丰富，并且 星形胶质细胞GQ信号的激活足以促进清醒。虽然星形胶质细胞至关重要 通过嘌呤能信号传导中介导其他大脑区域的突触传播，神经元的作用 乙醇暴露期间VPAG中的星形胶质细胞相互作用需要进一步投资。这个F30旨在 测试中心假设，即来自相邻星形胶质细胞的嘌呤能传播介导 VPAGDA神经元的去肾上腺素能调节，乙醇增强了该信号传导。目标是 阐明机械细节介导兴奋驱动器上唤醒vpagda的增加 通过激活α1AR的神经元和急性后去甲肾上腺素能系统的病理变化 慢性酒精暴露。在特定目标1中，我将使用药物和病毒遗传方法 结合离体荧光成像和全细胞贴片钳电生理学，以检查 VPAGDA神经元的甲肾上腺素能调节基础的神经元 - 核细胞相互作用。在特定的目标2中，我会 除了慢性间歇性乙醇暴露的小鼠模型外，还使用类似的方法来确定 急性和慢性乙醇暴露后突触传播到VPAGDA神经元的变化。这 这些研究的结果将使我们的实验室，我们的合作者和其他人进一步研究神经元的作用 与酒精使用障碍的睡眠功能障碍有关的星形胶质细胞生理学。这将进一步迈向 确定可能开发新疗法的治疗靶点，以减轻障碍的睡眠 并有助于预防救济。