Neuronal mechanisms mediating the effects of chronic alcohol consumption on sleep homeostasis.

介导长期饮酒对睡眠稳态影响的神经机制。

• 批准号: 10470383
• 负责人: MAHESH M THAKKAR
• 金额: $ 36.29万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-20 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10470383
• 关键词: Abstinence; Acute; Adenosine A1 Receptor; Adenosine Kinase; Affect; Alcohol consumption; Alcohol dependence; Alcoholism; Alcohols; Anti-Inflammatory Agents; Attenuated; B-Cell Activation; Brain; Brain Diseases; Cessation of life; Chronic; Coupled; Electrophysiology (science); Enhancers; Enterobacteria phage P1 Cre recombinase; Equilibrative Nucleoside Transporter 1; Etiology; Exposure to; Functional disorder; Gene Expression; Genes; Gliosis; Histone Deacetylase Inhibitor; Histones; Homeostasis; Human; Image; Impairment; Inflammatory; Infusion procedures; Interleukin-1 beta; Interleukins; Internal Ribosome Entry Site; Knock-in; Lesion; Light; MAP Kinase Gene; Mediating; Mediator of activation protein; Microdialysis; Minocycline; Mus; NOS2A gene; National Institute on Alcohol Abuse and Alcoholism; Neurons; Nuclear; Nucleoside Transporter; Patients; Pattern; Pharmacogenetics; Pharmacology; Play; Polysomnography; Relapse; Research; Role; Severities; Signal Transduction; Sleep; Sleep Disorders; Sleep disturbances; Sleeplessness; Symptoms; System; TNF gene; Techniques; Technology; Testing; Time; Toll-like receptors; Transgenic Organisms; Trichostatin A; Up-Regulation; Viral; Wakefulness; alcohol effect; alcohol use disorder; awake; basal forebrain; basal forebrain cholinergic neurons; cholinergic; cholinergic neuron; chronic alcohol ingestion; clinically relevant; cytokine; design; designer receptors exclusively activated by designer drugs; disability; drinking; efficacious treatment; epigenome; improved; in vivo; in vivo calcium imaging; indexing; inhibitor; innovation; kinase inhibitor; multidisciplinary; neuroinflammation; non rapid eye movement; novel; overexpression; premature; prevent; problem drinker; programs; receptor; relapse prediction; sleep quantity; treatment strategy

Alcohol use disorder (AUD), a chronic relapsing brain disorder, characterized by compulsive and excessive alcohol use, is a leading preventable cause of premature disability and death. Insomnia and associated sleep disturbances are amongst the most severe and protracted symptoms of AUD. In fact, subjective and objective indicators of sleep disturbances are predictors of relapse. Thus, while there is a direct relationship between insomnia and AUD, the underlying pathophysiology is not well understood. Evidence suggest that sleep homeostatic mechanism [adenosinergic/cholinergic mechanisms in the basal forebrain (BF)] may play a crucial role in mediating the effects of alcohol on sleep. Hence, we hypothesize that chronic alcohol consumption directly and indirectly, via neuroinflammation, disrupts sleep homeostasis leading to insomnia and sleep disturbances. We will use wild type (C57BL/6J) and transgenic ChAT-cre (expression of Cre-recombinase exclusively in cholinergic neurons) mice and expose them to the common maladaptive pattern of alcohol consumption: chronic intermittent access two-botte choice paradigm (IA2BC). A novel combination of multidisciplinary techniques including in vivo calcium imaging, pharmacogenetics, viral-mediating gene overexpression, microdialysis and local pharmacological manipulations will be used. Three aims are designed to test our hypothesis. In Aim 1, live Ca+2 imaging coupled with electrophysiological monitoring of sleep-wakefulness will be performed to examine the relationship between cholinergic neuronal activity, wakefulness and chronic alcohol intake. We predict that mice exposed to IA2BC will consume alcohol in an escalating pattern and display a positive relationship between cholinergic neuronal activity, disrupted sleep homeostasis and severity of sleep disruptions. While pharmacogenetic silencing of BF cholinergic neurons will attenuate the effects of chronic alcohol and normalize sleep, pharmacogenetic activation will attenuate sleep-promoting effects of acute alcohol. Aim 2 will examine the effects of chronic alcohol consumption on sleep homeostasis. We predict that 1) alcohol will downregulate MAPK signaling cascade and acetylated histones, to reduce the expressions of equilibrative nucleoside transporter 1 (ENT1) and adenosine A1 receptor (A1R) in the BF. 2) Virally mediated overexpression of ENT1 in the BF will attenuate the effects of alcohol on sleep homeostasis and improve the quality and quantity of sleep. 3) Local infusion of trichostatin-A (histone deacetylase inhibitor) will normalize sleep by upregulation of acetylated histones along with the expression of ENT1 and A1R in the BF, without affecting MAPK signaling. Aim 3 will examine chronic alcohol induced neuroinflammation in the BF and its effect on sleep homeostasis. Our predictions are that selective blockade of 1) neuroinflammatory changes by minocycline, 2) Toll-like receptors by TAK-242 and/or c) adenosine kinase by ABT-702, in the BF will attenuate chronic alcohol induced neuroinflammation; reduce gliosis and cytokine levels in the BF and normalize the indices of sleep homeostasis and sleep-wakefulness.

酒精使用障碍（AUD）是一种慢性复发性脑功能障碍，以强迫和过度饮酒为特征。 酗酒是导致过早残疾和死亡的主要可预防原因。失眠和相关睡眠 这些紊乱是AUD最严重和最持久的症状之一。事实上，主观和客观 睡眠障碍的指标是复发的预测因子。因此，虽然两者之间存在直接关系， 失眠和AUD，潜在的病理生理学还没有很好地理解。有证据表明睡眠 稳态机制[基底前脑（BF）中的腺苷能/胆碱能机制]可能在脑内的代谢中起关键作用。 在调节酒精对睡眠的影响中的作用。因此，我们假设长期饮酒直接 并通过神经炎症间接破坏睡眠稳态，导致失眠和睡眠障碍。 我们将使用野生型（C57 BL/6 J）和转基因ChAT-cre（Cre重组酶仅在大肠杆菌中表达）。 胆碱能神经元）小鼠，并将其暴露于常见的适应不良的饮酒模式：慢性 间歇性访问两瓶选择范式（IA 2BC）。多学科技术的新组合 包括体内钙成像、药物遗传学、病毒介导的基因过表达、微透析和 将使用局部药理学操作。我们设计了三个目标来检验我们的假设。 在目标1中，将进行活体Ca+2成像结合睡眠-觉醒的电生理监测 研究胆碱能神经元活动、觉醒和慢性酒精摄入之间的关系。我们 预测暴露于IA 2BC的小鼠将以逐步升级的模式消耗酒精，并显示出阳性反应。 胆碱能神经元活动、睡眠稳态紊乱和睡眠紊乱严重程度之间的关系。 而BF胆碱能神经元的药物遗传学沉默将减弱慢性酒精的影响， 正常化睡眠，药物遗传学激活将减弱急性酒精的睡眠促进作用。 目的2将研究长期饮酒对睡眠稳态的影响。我们预测1）酒精 将下调MAPK信号级联和乙酰化组蛋白，减少平衡蛋白的表达， BF中的核苷转运蛋白1（ENT 1）和腺苷A1受体（A1 R）。2)病毒介导的过表达 在BF中ENT 1的表达将减弱酒精对睡眠稳态的影响，并改善睡眠的质量和数量。 睡眠。3)局部输注阿司他汀-A（组蛋白去乙酰化酶抑制剂）将通过上调 乙酰化组蛋白沿着ENT 1和A1 R在BF中的表达，而不影响MAPK信号传导。 目的3将研究慢性酒精诱导的BF神经炎症及其对睡眠稳态的影响。 我们的预测是：1）米诺环素选择性阻断神经炎性变化，2）Toll样 受体和/或c）ABT-702的腺苷激酶，将减弱BF中慢性酒精诱导的 神经炎症;减少BF中的神经胶质增生和细胞因子水平，并使睡眠稳态指数正常化 和睡眠-觉醒。