Dependence-Induced Excessive Ethanol Consumption: Role of Corticostriatal Kv7 Channels

摘要 这是一份申请导师研究科学家发展奖(K01)以支持职业生涯的申请书 Jennifer Rinker博士的发展和强化培训，以促进她向独立学者的过渡 酒精研究领域的调查员。候选人是一名早期调查员，正在从 博士后研究员至助理教授。林克博士在体内药理学和 研究酒精使用障碍所涉及的神经回路的化学遗传学操作，但有限 有切片电生理学和先进的分子技术检查可塑性相关事件的经验。 已经制定了一项密集和全面的培训、指导和研究计划，将提供 高级技术培训以评估KV7通道参与酒精依赖诱导的改变 功能可塑性。林克博士的培训将得到对她职业生涯的坚定机构承诺的支持 酒精研究领域的开发和强大的领导指导团队，每个人都提供战略 随着事业的发展，她在这项提案的制定和指导方面都提供了指导。建议数 研究计划是林克博士最近在导师医院进行的研究的自然延伸 实验室，但区别在于它在调制中检查离散皮质纹状体回路中的KV7通道 依赖诱导的酒精消费的影响。内侧前额叶皮质(MPFC)是一个关键的 涉及对奖赏动机行为和投射到核团施加抑制控制的结构 伏隔核(NAC)是中脑边缘奖赏通路的重要组成部分。酒精依赖是 与过量和无节制的饮酒有关，已知会改变mPFC的可塑性和生理学 锥体神经元。具体地说，乙醇戒断导致NAC投射的mPFC的过度兴奋。 神经元，其潜在的机制尚不清楚。KV7通道产生关键的M-电流 通过维持膜电位和抑制神经元放电来调节神经元的兴奋性。这些 通道参与调节NAC中的乙醇消耗，但它们在皮质纹状体中的作用 依赖的啮齿类动物体内的回路仍未被探索。因此，这一提议的首要假设是 依赖诱导皮质纹状体回路中KV7通道的神经适应(即mPFC到NAC) 依赖于推动乙醇消费升级和失控。这项提案将检验这一点 使用多方面方法结合蛋白质表达和分析的亚细胞分析的假说 分别使用Diolistic标记和切片电生理学对结构和功能可塑性进行研究。我们的结果 大量饮酒和依赖小鼠中KV7通道的参与表明 这些研究将极大地促进我们对乙醇背后的细胞机制的理解。 依赖。此机会将为应聘者提供全面的培训和坚实的基础 从而在酒精神经科学领域建立一个成功和独立的研究项目。

ABSTRACT This is an application for a Mentored Research Scientist Development Award (K01) to support the career development and intensive training of Dr. Jennifer Rinker to facilitate her transition to an independent academic investigator in the alcohol research field. The candidate is an early-stage investigator transitioning from Postdoctoral Fellow to Assistant Professor. Dr. Rinker has extensive experience with in vivo pharmacology and chemogenetic manipulations to study the neural circuitry involved in alcohol use disorder, but has limited experience with slice electrophysiology and advanced molecular techniques to examine plasticity-related events. An intense and comprehensive training, mentoring and research plan has been developed that will provide training in advanced techniques to assess Kv7 channel involvement in alcohol dependence-induced changes in functional plasticity. Dr. Rinker's training will be supported by a firm institutional commitment to her career development and a strong mentoring team of leaders in the alcohol research field, each providing strategic guidance in both the development of this proposal and mentoring as her career progresses. The proposed research plan is a natural extension of the recent studies Dr. Rinker has been conducting in the mentor's laboratory, but is distinguished by its examination of Kv7 channels in discrete corticostriatal circuits in modulating the effects of dependence-induced ethanol consumption. The medial prefrontal cortex (mPFC) is a critical structure involved in imposing inhibitory control over reward-motivated behaviors and projects to the nucleus accumbens (NAc), an essential component of the mesolimbic reward pathway. Ethanol dependence is associated with elevated and uncontrolled drinking and is known to alter the plasticity and physiology of mPFC pyramidal neurons. Specifically, ethanol withdrawal results in the hyperexcitability of NAc-projecting mPFC neurons, the underlying mechanism of which remains unknown. Kv7 channels generate the M-current that critical regulates neuronal excitability by maintaining the membrane potential and dampening neuronal firing. These channels have been implicated in regulating ethanol consumption in the NAc, but their role in the corticostriatal circuits in dependent rodents remains unexplored. Thus, the overarching hypothesis of this proposal is that dependence-induced neuroadaptations in Kv7 channels in the corticostriatal circuitry (i.e., mPFC to NAc) drive the escalated and uncontrolled ethanol consumption in dependence. This proposal will test this hypothesis using a multifaceted approach incorporating subcellular analysis of protein expression and analysis of structural and functional plasticity using diolistic labeling and slice electrophysiology, respectively. Our results demonstrating involvement of Kv7 channels in heavy drinking and dependent mice suggests that continuing these studies will significantly advance our understanding of the cellular mechanisms underlying ethanol dependence. This opportunity will provide the candidate with comprehensive training and a solid foundation on which to build a successful and independent research program in the alcohol neuroscience field.