Exploring Neuronal Subtype-specific Mechanisms of Ethanol Dependence and Reward: Contributions of Voltage-gated Potassium Channels

探索乙醇依赖和奖励的神经元亚型特异性机制：电压门控钾通道的贡献

• 批准号: 10395716
• 负责人: Reginald DeVon Cannady
• 金额: $ 24.9万
• 依托单位: NORTH CAROLINA AGRI & TECH ST UNIV
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-05 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10395716
• 关键词: Air; Alcohol consumption; Alcohol dependence; Alcohols; Animal Model; Animals; Automobile Driving; Behavior; Brain; Cell physiology; Chemosensitization; Chronic; Cues; Data; Dependence; Development; Dopamine Receptor; Electrophysiology (science); Ethanol; Ethanol dependence; Exposure to; Extinction (Psychology); Family; Female; Fiber; Functional disorder; Genes; Heavy Drinking; Home; Individual; Infusion procedures; Interneuron function; Interneurons; Laboratories; Life; Link; Measures; Medial; Mediating; Modeling; Motivation; Mouse Strains; Mus; Neurons; Operant Conditioning; Parvalbumins; Phase; Photometry; Potassium Channel; Prefrontal Cortex; Procedures; Property; Publishing; Rattus; Regulation; Relapse; Research Project Grants; Rewards; Rodent Model; Role; Self Administration; Signal Transduction; Specificity; Sucrose; Surface; Techniques; Testing; Time; Training; Transcript; Transgenic Animals; Transgenic Mice; Transgenic Organisms; Viral; Voltage-Gated Potassium Channel; Western Blotting; Withdrawal Symptom; alcohol cue; alcohol effect; alcohol exposure; alcohol reinforcement; alcohol relapse; alcohol response; alcohol reward; alcohol seeking behavior; alcohol use disorder; cognitive function; drinking; drug seeking behavior; effective therapy; experimental study; hippocampal pyramidal neuron; in vivo; inhibitory neuron; insight; interest; knock-down; male; neuroadaptation; neurotransmission; new technology; novel; patch clamp; relapse risk; therapeutic target; vapor; voltage

Project Summary/ Abstract Individuals with alcohol use disorder (AUD) show deficits in cognitive function and an inability to regulate ethanol consumption and seeking. These deficits are driven by medial prefrontal cortex (mPFC) dysfunction. Ethanol dependence induced by chronic intermittent ethanol (CIE) vapor exposure disrupts expression of mPFC Kcnc1. This gene encodes voltage-dependent KV3.1 potassium channels, which are highly enriched in parvalbumin positive fast-spiking interneurons (PV+FSIs). Our preliminary data show that CIE-induced dysregulation of mPFC PV+FSI activity is restored by a novel KV3 channel positive modulator which also reduced drinking in dependent and non-dependent mice. These data implicate aberrant KV3 channel activity in PV+FSIs with dependence- induced deficits in mPFC function. We hypothesize that dependence-induced excessive drinking, ethanol reinforcement, and cue-induced relapse-like behavior are regulated by disruption of mPFC KV3 channel activity in PV+FSIs. Specific Aim 1 will examine functional neuroadaptations of KV3 channel activity in mPFC PV+FSIs of G42 (PV-GFP) transgenic mice exposed to CIE or Air, and test the effects of KV3 positive modulators to restore dependence-induced adaptations in PV+FSI function. We will also test if reduced KV3 channel expression is an underlying cellular mechanism driving aberrant KV3 channel activity in PV+FSIs. Specific Aim 2 will test if enhancing KV3 channel activity in the mPFC will reduce ethanol consumption in CIE and Air-exposed mice during home cage drinking sessions. Studies will also determine if KV3 positive modulation can restore mPFC PV+FSI activity during home cage drinking sessions in PV-Cre mice using in vivo fiber photometry techniques. Specific Aim 3 (R00 phase) will shift from studying home cage drinking to studying the role of mPFC KV3 channels in regulating the reinforcing effects of ethanol and relapse-like behavior by using operant conditioning procedures in dependent and non-dependent rats. Using in vivo fiber photometry, studies will test if positive modulation of KV3 increases Ca2+ transients in PV+FSIs in CIE-exposed PV-Cre rats, and in turn, decreases motivation to self-administer ethanol. Experiments will also determine if enhanced KV3 activity reduces ethanol-seeking behavior and modifies Ca2+ transients in PV+FSIs during cue-induced reinstatement tests in CIE-exposed PV-Cre rats. Collectively, these studies will validate KV3 channels as a novel target for the development of effective treatment options for ethanol dependence. In addition, the proposed experiments will provide training in emerging techniques used to selectively measure cellular function in sub-populations of cortical neurons of behaving transgenic animals.

项目概要/摘要 患有酒精使用障碍 (AUD) 的个体表现出认知功能缺陷和无法调节乙醇 消费和寻求。这些缺陷是由内侧前额叶皮层 (mPFC) 功能障碍引起的。乙醇 慢性间歇性乙醇 (CIE) 蒸气暴露引起的依赖性会破坏 mPFC Kcnc1 的表达。 该基因编码电压依赖性 KV3.1 钾通道，该通道富含小清蛋白 正快速尖峰中间神经元（PV+FSIs）。我们的初步数据表明，CIE 引起的 mPFC 失调 PV+FSI 活性通过新型 KV3 通道正调节剂恢复，这也减少了依赖性饮酒 和非依赖性小鼠。这些数据暗示 PV+FSI 中异常的 KV3 通道活动具有依赖性 诱发 mPFC 功能缺陷。我们假设依赖引起的过量饮酒、乙醇 强化和提示诱导的复发样行为是通过破坏 mPFC KV3 通道活性来调节的 在光伏+金融服务机构中。具体目标 1 将检查 mPFC PV+FSIs 中 KV3 通道活动的功能性神经适应 G42 (PV-GFP) 转基因小鼠暴露于 CIE 或空气，并测试 KV3 阳性调节剂对 恢复 PV+FSI 功能中依赖性诱导的适应。我们还将测试是否减少KV3通道 表达是驱动 PV+FSIs 中异常 KV3 通道活性的潜在细胞机制。具体目标 2 将测试增强 mPFC 中的 KV3 通道活性是否会减少 CIE 和空气暴露中的乙醇消耗 老鼠在家里的笼子里喝酒。研究还将确定 KV3 正调节是否可以恢复 使用体内光纤光度测定法对 PV-Cre 小鼠在家笼饮酒期间 mPFC PV+FSI 活性 技术。具体目标 3（R00 阶段）将从研究家庭笼内饮酒转向研究 mPFC 的作用 KV3 通道通过操作调节乙醇的强化作用和复发样行为 依赖性和非依赖性大鼠的调节程序。研究将使用体内光纤光度测定法来测试是否 KV3 的正向调节会增加暴露于 CIE 的 PV-Cre 大鼠中 PV+FSI 中的 Ca2+ 瞬变，进而， 降低自我施用乙醇的动力。实验还将确定 KV3 活性是否增强 减少乙醇寻找行为并改变提示诱导恢复期间 PV+FSI 中的 Ca2+ 瞬变 在 CIE 暴露的 PV-Cre 大鼠中进行测试。总的来说，这些研究将验证 KV3 通道作为 开发乙醇依赖的有效治疗方案。此外，拟议的实验将 提供用于选择性测量亚群细胞功能的新兴技术的培训 行为转基因动物的皮质神经元。