Role of afferents to the rostromedial tegmental nucleus in symptoms of withdrawal from chronic ethanol exposure

头内侧被盖核传入神经在慢性乙醇暴露戒断症状中的作用

• 批准号: 10750827
• 负责人: Hyerim Yang
• 金额: $ 4.47万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-16 至 2025-06-15
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10750827
• 关键词: Abstinence; Acute; Affect; Alcohol consumption; Alcohol withdrawal syndrome; Animals; Anxiety; Attenuated; Behavioral; Brain region; Cell Nucleus; Cells; Chronic; Clinical; CpG Islands; DNA; DNA Methylation; Data; Electrophysiology (science); Epigenetic Process; Ethanol; Fluorescent in Situ Hybridization; Gene Expression; Genes; Genetic; Genetic Transcription; Glutamates; Habenula; Human; Hyperactivity; Hyperalgesia; Hypermethylation; Individual; Lateral; Measures; Mechanics; Mediating; Mental Depression; Methylation; Molecular; Neurons; Neurosciences; Neurosciences Research; Pain; Pharmacology; Physiological; Plasma; Play; Predisposition; Promoter Regions; Publishing; Rattus; Relapse; Role; Scientist; Signal Transduction; Sleep disturbances; Slice; Symptoms; Techniques; Testing; Training; Transcriptional Regulation; Tremor; Withdrawal; Withdrawal Symptom; Work; addiction; alcohol abuse therapy; alcohol behavior; alcohol exposure; alcohol use disorder; anxiety-like behavior; avoidance behavior; career; epigenetic regulation; experience; experimental study; genetic approach; in vivo; innovation; insight; mRNA Expression; mu opioid receptors; negative affect; neural; neuroadaptation; neuromechanism; neurotransmission; new therapeutic target; pain sensitivity; patch clamp; pharmacologic; pre-clinical; promoter; receptor; sobriety; withdrawal-induced anxiety

ABSTRACT A significant number of individuals with alcohol use disorder struggle to maintain sobriety as a result of withdrawal symptoms that emerge during early abstinence. Despite this, the neural mechanisms underlying symptoms of withdrawal are not well understood. The rostromedial tegmental nucleus (RMTg) is a GABAergic region that plays a critical role in avoidance behavior, aversive signaling, pain, and importantly, alcohol-related behaviors. Recent work from our lab provides evidence of RMTg hyperactivity during acute withdrawal from chronic intermittent ethanol (CIE) exposure. In addition, we showed that pharmacological inhibition of the RMTg attenuates withdrawal-induced anxiety-like behavior. The afferents that drive RMTg-mediated withdrawal symptoms are currently unknown. However, previously published studies have implicated the lateral habenula (LHb) – a region that provides dense input to the RMTg – in withdrawal-induced negative affect similar to our work in the RMTg. In addition, our preliminary data revealed significant cFos induction in RMTg-projecting LHb neurons during acute withdrawal in CIE-exposed rats compared to AIR controls. Interestingly, the LHb is heavily enriched in mu-opioid receptors (MORs), which are known to regulate both pain and affect. Oprm1, the gene that encodes MOR, is hypermethylated in individuals with AUD suggestive of a potential mechanism by which chronic ethanol exposure drives decreases in Oprm1 expression. However, the effect of withdrawal on MORs in the LHb is unclear. Together, these data lead us to hypothesize that LHb afferents to the RMTg are mechanistically involved in withdrawal from chronic ethanol and that epigenetic dysregulation of the Oprm1 gene may contribute to this mechanism. Three specific aims will be used to test this hypothesis. In Aim 1, in vivo chemogenetics will be used to determine whether selective inhibition of RMTg-projecting LHb neurons reverses symptoms of withdrawal. Oprm1 expression and methylation levels will be assessed in RMTg-projecting LHb neurons in Aim 2 using fluorescent in situ hybridization and MethylMiner. Tract tracing will be combined with ex vivo whole-cell patch-clamp slice electrophysiology in Aim 3 to investigate withdrawal-induced changes in epigenetic regulation of MOR receptor-mediated firing in RMTg-projecting LHb neurons. These studies will provide the applicant with new training in cutting-edge neuroscience techniques while also providing new insight into the role of the LHb-RMTg circuit in ethanol withdrawal.

抽象的 大量的酒精使用障碍患者因退出而努力保持清醒 早期禁欲时出现的症状。尽管如此，神经机制的潜在症状 戒断不太了解。 roust骨技术核（RMTG）是一个GABA能区域，该区域 在回避行为，厌恶信号传导，痛苦以及与酒精相关的行为中起着至关重要的作用。 我们实验室的最新工作提供了急性退出慢性期间RMTG多动症的证据 间歇性乙醇（CIE）暴露。此外，我们表明了RMTG的药物抑制 减弱戒断引起的类似动画的行为。驱动RMTG介导的退出的传入 症状目前未知。但是，以前发表的研究已经实施了侧向habenula （LHB） - 一个为RMTG提供密集输入的区域 - 在撤回引起的负面影响与我们 在RMTG中工作。此外，我们的初步数据显示RMTG投影LHB的CFO诱导显着 与空气对照相比，暴露于CIE的大鼠急性戒断期间的神经元。有趣的是，LHB很重 富含MU-阿片受体（MOR），已知可以调节疼痛和影响。 OPRM1，基因 编码MOR，在具有AUD的个体中是高甲基化的，暗示了潜在的机制 慢性乙醇暴露在OPRM1表达中驱动下降。但是，退出对MOR的影响 LHB不清楚。这些数据一起导致我们假设LHB的传入RMTG是 机械上参与退出慢性乙醇的戒断和OPRM1基因的表观遗传失调 可能有助于这种机制。将使用三个特定目标来检验这一假设。在AIM 1中，体内 化学遗传学将用于确定选择性抑制RMTG投射LHB神经元是否反向 戒断的症状。 OPRM1表达和甲基化水平将在RMTG投影LHB中评估 AIM 2中的神经元使用荧光原位杂交和甲基分子。道跟踪将与Ex结合 AIM 3中的Vivo全细胞贴剂切片电生理学，以研究戒断诱导的变化 RMTG投射LHB神经元中MOR受体介导的发射的表观遗传调节。这些研究会 为申请人提供尖端神经科学技术的新培训，同时还提供新的见解 进入LHB-RMTG电路在乙醇提取中的作用。