Ethanol and mGluR2 signaling

乙醇和 mGluR2 信号传导

• 批准号: 10745067
• 负责人: Rong Chen
• 金额: $ 55.67万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-20 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10745067
• 关键词: 3-hydroxy-3-methylglutaryl-coenzyme A; Ablation; Acceleration; Affect; Alcohol consumption; Alcohols; Animals; Attenuated; Behavior; Behavioral; Binding; Brain; Brain region; CRISPR/Cas technology; Cholesterol; Cholesterol Homeostasis; Chronic; Data; Density Gradient Centrifugation; Dependence; Electrophysiology (science); Enzymes; Ethanol; Ethanol dependence; Excision; GTP-Binding Proteins; Genes; Glutamates; Impairment; Knowledge; Learning; Lentivirus Vector; Measures; Mediating; Membrane; Microdialysis; Molecular; Molecular Target; Monitor; Mood Disorders; National Institute on Alcohol Abuse and Alcoholism; Neurobiology; Neurons; Oxidoreductase; Patients; Prefrontal Cortex; Rattus; Reporting; Research; Rodent Model; Signal Transduction; Strategic Planning; Sucrose; Synapses; Testing; Therapeutic Intervention; Tissues; alcohol abuse therapy; alcohol behavior; alcohol comorbidity; alcohol exposure; alcohol use disorder; anxiety-like behavior; behavior measurement; extracellular; improved; innovation; metabotropic glutamate receptor 2; neural; neuronal excitability; neurophysiology; neurotransmission; novel; postsynaptic; presynaptic; prevent; protein activation; segregation; tool; transmission process; 3-hydroxy-3-methylglutaryl-coenzyme A; Ablation; Acceleration; Affect; Alcohol consumption; Alcohols; Animals; Attenuated; Behavior; Behavioral; Binding; Brain; Brain region; CRISPR/Cas technology; Cholesterol; Cholesterol Homeostasis; Chronic; Data; Density Gradient Centrifugation; Dependence; Electrophysiology (science); Enzymes; Ethanol; Ethanol dependence; Excision; GTP-Binding Proteins; Genes; Glutamates; Impairment; Knowledge; Learning; Lentivirus Vector; Measures; Mediating; Membrane; Microdialysis; Molecular; Molecular Target; Monitor; Mood Disorders; National Institute on Alcohol Abuse and Alcoholism; Neurobiology; Neurons; Oxidoreductase; Patients; Prefrontal Cortex; Rattus; Reporting; Research; Rodent Model; Signal Transduction; Strategic Planning; Sucrose; Synapses; Testing; Therapeutic Intervention; Tissues; alcohol abuse therapy; alcohol behavior; alcohol comorbidity; alcohol exposure; alcohol use disorder; anxiety-like behavior; behavior measurement; extracellular; improved; innovation; metabotropic glutamate receptor 2; neural; neuronal excitability; neurophysiology; neurotransmission; novel; postsynaptic; presynaptic; prevent; protein activation; segregation; tool; transmission process

Project Summary Current NIAAA strategic plan calls for efforts to identify novel molecular actions of alcohol and new remedies to mitigate alcohol-impacted brain regions. Ever-changing research tools have exponentially accelerated our understanding of brain regions and circuits affected by alcohol use and comorbid affective disorders. However, efforts towards the understanding of molecular mechanisms for altered neural activity in these impacted regions have lagged behind. To fill in this gap, we will explore a previously unknown molecular mechanism whereby ethanol modulation of brain cholesterol in the prefrontal cortex (PFC) leads to aberrant mGluR2 function and glutamate transmission in a rodent model of chronic intermittent ethanol (CIE) exposure. This modulation may thus contribute to alcohol drinking and anxiety-like behavior, commonly observed in patients with alcohol use disorder (AUD). Using a CIE rodent model, we show that CIE increases glutamate release in the PFC along with increased alcohol drinking and anxiety-like behavior. Further, we and others report that ablation of PFC neuron projection to subcortical regions (e.g. BLA or NAc) abolishes these behaviors. Thus, the PFC is a critical hub for regulating dependence-related neural and behavioral activity. However, the molecular mechanisms underlying CIE-induced disruption of glutamate transmission in the PFC remain elusive. Our preliminary data strongly suggest mGluR2 involvement in this region: a) CIE spatially segregates mGluR2 from Gαo subunit and reduces mGluR2 stimulation of Gαi/o; b) CIE abolishes mGluR2 inhibition of Gβγ-mediated presynaptic glutamate release and postsynaptic intrinsic excitability of PFC neurons; and c) CIE impairs mGluR2 inhibition of anxiety-like behavior. Such a deficit in mGluR2 function may be governed by membrane cholesterol. We found that CIE drastically increases cholesterol content in the PFC by enhancing the activity of HMG-CoA-reductase (HMGCR), a cholesterol synthesis enzyme. Importantly, reduced, CIE- dependent mGluR2 function can be reversed by ex vivo cholesterol removal and mimicked by ex vivo cholesterol addition to naïve tissue. We hypothesize that blockade of cholesterol increase will attenuate CIE- dependent disruption of mGluR2-mediated G-protein activation, neurophysiology and dependence-related behaviors. We will use CRISPR/Cas9, cellular, electrophysiological and behavioral approaches to test our hypothesis. The proposal is conceptually and technically innovative and significant, investigating a largely overlooked and potentially impactful effect of CIE exposure on cholesterol modulation of mGluR2 function, glutamate transmission and anxiety-like behavior.

项目摘要 当前的NIAAA战略计划要求努力确定酒精和新疗法的新分子作用 减轻受酒精影响的大脑区域。不断变化的研究工具已成倍地加速了我们的 了解受酒精使用和合并情感障碍影响的大脑区域和电路。然而， 在这些影响中了解改变神经活动的分子机制的努力 地区落后了。要填补这一空白，我们将探索以前未知的分子机制 乙醇调节前额叶皮层（PFC）的脑胆固醇会导致异常MGLUR2 慢性间歇性乙醇（CIE）暴露的啮齿动物模型中的功能和谷氨酸传播。这 因此，调节可能有助于饮酒和类似动画的行为，通常在患者中观察到 患有酒精疾病（AUD）。使用CIE啮齿动物模型，我们表明CIE增加了谷氨酸的释放 PFC以及饮酒和类似焦虑的行为的增加。此外，我们和其他人报告了 PFC神经元对皮层下区域（例如BLA或NAC）的消融可取消这些行为。那， PFC是控制与依赖性相关的神经和行为活动的关键枢纽。但是， CIE诱导的PFC中谷氨酸传播的破坏的分子机制仍然存在 难以捉摸。我们的初步数据强烈建议MGLUR2参与该区域：a）CIE在空间上分离 来自GαO亚基的mglur2并减少了GαI/O的mglur2刺激； b）CIE废除mglur2抑制 Gβγ介导的PFC神经元的突触前谷氨酸释放和突触后内在的刺激性；和C）CIE 损害MGLUR2对动画样行为的抑制。 MGLUR2功能的这种赤字可能由 膜胆固醇。我们发现CIE通过增强PFC大大增加了PFC中的胆固醇含量 HMG-COA-还原酶（HMGCR）的活性，一种胆固醇合成酶。重要的是，减少，cie- 依赖的mglur2功能可以通过离体胆固醇去除并通过ex vivo逆转 胆固醇添加到幼稚的组织中。我们假设对胆固醇升高的封锁将减轻CIE- MGLUR2介导的G蛋白激活，神经生理学和与依赖性相关的依赖性破坏 行为。我们将使用CRISPR/CAS9，细胞，电生理和行为方法来测试我们的 假设。该提案在概念上和技术上都是创新且重要的，在很大程度上调查了 CIE暴露对MGLUR2功能调节的忽视和潜在影响的影响， 谷氨酸传播和类似动画的行为。