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。

基本信息

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
项目类别：
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

项目摘要

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），一种慢性复发脑障碍，其特征是强迫性和过度饮酒是过早残疾和死亡的可预防原因。失眠和相关的睡眠干扰是AUD最严重和最旷日持久的症状之一。实际上，主观和客观睡眠障碍的指标是复发的预测指标。因此，虽然之间存在直接关系失眠和AUD，基本的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]可能发挥至关重要的在介导酒精对睡眠的影响中的作用。因此，我们假设直接慢性酒精消耗通过神经炎症间接地破坏了睡眠体内平衡，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（仅在胆碱能神经元）小鼠并将其暴露于饮酒的常见不良适应性模式：慢性间歇性访问两底选择范式（IA2BC）。多学科技术的新型组合包括体内钙成像，药物遗传学，病毒中性基因过表达，微透析和将使用局部药理操作。三个目标旨在检验我们的假设。在AIM 1中，将进行活睡眠的现场CA+2成像以及对睡眠的电生理监测检查胆碱能神经元活性，清醒和慢性酒精摄入之间的关系。我们预测暴露于IA2BC的小鼠将以升级的方式食用酒精并显示阳性胆碱能神经元活动，睡眠稳态中断和睡眠干扰的严重程度之间的关系。 BF胆碱能神经元的药物遗传沉默将减轻慢性酒精和使睡眠正常化，药物遗传学激活会减轻急性酒精的促进睡眠作用。 AIM 2将检查慢性酒精消耗对睡眠体内平衡的影响。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）病毒介导的过表达 BF中的ENT1的摄入会减弱酒精对睡眠体内平衡的影响并提高质量和数量睡眠。 3）局部输注trichostatin-A（组蛋白脱乙酰基酶抑制剂）将通过上调使睡眠正常化乙酰化组蛋白以及BF中ENT1和A1R的表达，而不会影响MAPK信号传导。 AIM 3将检查慢性酒精引起的BF中的神经炎症及其对睡眠体内平衡的影响。我们的预测是1）米诺环素的神经炎症变化的选择性封锁，2）tollike TAK-242和/或C）ABT-702的腺苷激酶的受体将减弱慢性酒精诱导神经炎症；降低BF的神经胶质病和细胞因子水平，并使睡眠体内稳态指标归一化和睡眠消失。