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: 10019446
• 负责人: MAHESH M THAKKAR
• 金额: $ 36.35万
• 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/10019446
• 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; epigenome; improved; in vivo; in vivo calcium imaging; indexing; inhibitor/antagonist; 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），一种慢性复发性脑部疾病，其特征是强迫性和过度饮酒 饮酒是导致过早残疾和死亡的一个可预防的主要原因。失眠和相关睡眠 紊乱是 AUD 最严重、最持久的症状之一。其实主观和客观 睡眠障碍的指标是复发的预测因素。因此，虽然两者之间存在直接关系 失眠和 AUD 的潜在病理生理学尚不清楚。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]可能起着至关重要的作用 调节酒精对睡眠影响的作用。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 (C57BL/6J) 和转基因 ChAT-cre（Cre 重组酶仅在 胆碱能神经元）小鼠，并使它们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例（IA2BC）。多学科技术的新颖结合 包括体内钙成像、药物遗传学、病毒介导基因过度表达、微透析和 将使用局部药理学操作。设计了三个目标来检验我们的假设。 在目标 1 中，将进行实时 Ca+2 成像以及睡眠-觉醒的电生理监测 检查胆碱能神经元活动、觉醒和长期饮酒之间的关系。我们 预测暴露于 IA2BC 的小鼠会以逐渐增加的方式饮酒并表现出阳性 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。 虽然 BF 胆碱能神经元的药物遗传学沉默会减弱慢性酒精和酒精的影响 使睡眠正常化，药物遗传学激活会减弱急性酒精的促进睡眠作用。 目标 2 将检查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，以减少平衡的表达 BF 中的核苷转运蛋白 1 (ENT1) 和腺苷 A1 受体 (A1R)。 2) 病毒介导的过度表达 BF 中的 ENT1 会减弱酒精对睡眠稳态的影响并提高睡眠的质量和数量 睡眠。 3) 局部输注曲古抑菌素-A（组蛋白脱乙酰酶抑制剂）将通过上调睡眠正常化 乙酰化组蛋白以及 BF 中 ENT1 和 A1R 的表达，而不影响 MAPK 信号传导。 目标 3 将检查慢性酒精引起的 BF 神经炎症及其对睡眠稳态的影响。 我们的预测是，米诺环素选择性阻断 1）神经炎症变化，2）Toll 样 TAK-242 受体和/或 ABT-702 c) 腺苷激酶，在 BF 中会减弱慢性酒精诱导的 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。