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: 10401948
• 负责人: Reginald DeVon Cannady
• 金额: $ 24.66万
• 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/10401948
• 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.

项目摘要/摘要 患有饮酒障碍（AUD）的个体表现出认知功能缺陷，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）功能障碍驱动的。乙醇 由慢性间歇性乙醇（CIE）蒸气暴露引起的依赖性破坏了MPFC KCNC1的表达。 该基因编码依赖电压的Kv3.1钾通道，高度富集白细胞蛋白 阳性快速刺激性中间神经元（PV+FSI）。我们的初步数据表明，CIE诱导的MPFC失调 PV+FSI活性由新型的KV3通道正调制器恢复，该调制器也减少了依赖的饮酒 和非依赖小鼠。这些数据暗示PV+FSI中的异常KV3通道活性与依赖性 - 诱发MPFC功能的缺陷。我们假设依赖性引起的饮酒过量，乙醇 MPFC KV3通道活性的破坏来调节增强和提示引起的复发行为 在PV+FSI中。具体目标1将检查MPFC PV+FSI中KV3通道活性的功能神经适应 暴露于CIE或空气的G42（PV-GFP）转基因小鼠的摄 恢复依赖性诱导的PV+FSI函数的适应性。我们还将测试是否减少KV3通道 表达是一种驱动PV+FSI中驱动异常KV3通道活性的潜在细胞机制。具体目标 2将测试MPFC中的KV3通道活性是否会减少CIE的乙醇消耗量 在家笼饮水期间的老鼠。研究还将确定KV3阳性调制是否可以恢复 MPFC PV+FSI活性在PV-CRE小鼠中使用体内纤维光度法 技术。特定的目标3（R00阶段）将从研究家庭笼饮酒转变为研究MPFC的作用 KV3通道通过使用操作者来调节乙醇和复发行为的增强作用 依赖和非依赖大鼠的调节程序。使用体内纤维光度法，研究将测试是否 KV3的正调节增加了暴露于CIE的PV-CRE大鼠PV+ FSI中的Ca2+瞬变，然后 减少自我管理乙醇的动力。实验还将确定KV3活性是否增强 减少寻求乙醇的行为，并在提示诱导的恢复期间修改PV+ FSI中的Ca2+瞬变 暴露于CIE的PV-CRE大鼠中的测试。总的来说，这些研究将验证KV3通道作为新的目标 开发乙醇依赖的有效治疗选择。另外，提出的实验将 提供用于选择性测量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。