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

项目摘要/摘要 酒精使用障碍(AUD)患者表现出认知功能缺陷和无法控制酒精 消费和寻觅。这些缺陷是由内侧前额叶皮质(MPFC)功能障碍引起的。乙醇 慢性间歇性乙醇(CIE)蒸气暴露诱导的依赖破坏了mPFC KCNC1的表达。 这个基因编码电压依赖的KV3.1钾通道，它富含小白蛋白 快速放电中间神经元阳性(PV+FSIS)。我们的初步数据显示，CIE诱导的mPFC调节失调 一种新型的Kv3通道正性调节剂恢复了PV+FSI的活性，该调节剂也减少了依赖的饮酒 和非依赖的小鼠。这些数据提示在PV+FSIS中Kv3通道活动异常，并具有依赖性。 致mPFC功能缺陷。我们假设依赖导致过度饮酒，酒精 增强和提示诱导的复发样行为受mPFC Kv3通道活性破坏的调节 在PV+FSIS中。具体目标1将研究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或空气中，并检测Kv3正向调节剂对 恢复依赖诱导的PV+FSI功能适应。我们还将测试是否减少了Kv3频道 表达是导致PV+FSIS中Kv3通道活性异常的潜在细胞机制。特定目标 2将测试增强mPFC中Kv3通道的活性是否会减少CIE和暴露在空气中的乙醇消耗 老鼠在家中的笼子里喝水。研究还将确定Kv3正向调制是否可以恢复 体内纤维分光光度法检测PV-CRE小鼠家庭笼养饮酒过程中mPFC PV+FSI活性 技巧。具体目标3(R00阶段)将从研究家庭笼养饮酒转向研究mPFC的作用 Kv3通道通过操纵剂调节酒精强化效应和复吸样行为 依赖和非依赖大鼠的条件反射过程。使用体内纤维光度法，研究将测试是否 Kv3的正向调节增加了CIE暴露的PV-CRE大鼠PV+FSIS中的钙瞬变，进而， 降低自我给药乙醇的动力。实验还将确定增强的Kv3活性 在线索诱导的恢复过程中减少PV+FSIS的乙醇寻找行为和改变钙瞬变 在CIE暴露的PV-CRE大鼠身上进行测试。总的来说，这些研究将验证Kv3频道作为 开发有效的酒精依赖治疗方案。此外，拟议的实验将 提供新兴技术方面的培训，用于选择性地测量亚群中的细胞功能 转基因动物的大脑皮层神经元。

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.