Prefrontal Pathways Engaged in Excessive Alcohol Consumption

项目摘要 过度饮酒会对个人和社会产生广泛的影响， 同时造成重大的经济和法律的负担。尽管在过去的一段时间里， 几十年来，确定与发展相关的遗传影响和神经适应， 酒精使用障碍（AUDs），进展受到基础神经解剖学复杂性的限制。它 很明显，实验分辨率需要提高到识别特定适应性的水平。 与AUD病理相关的可分离行为的神经回路。因此，我的研究目标是 确定青少年酗酒导致的神经适应，使大量饮酒持续下去， 成年期的认知缺陷我的研究集中在前额叶皮层（PFC），因为这个区域 在青春期继续发展，可能容易受到大量饮酒的影响。比如说， 在酗酒者中观察到PFC功能障碍，可能导致强迫性饮酒， 在AUD中观察到认知缺陷。在老鼠身上，我发现青春期的狂饮会扰乱 在PFC依赖的工作记忆任务上的表现，增加成年后的酒精消费， 显著改变PFC锥体神经元的内在兴奋性。识别潜在的机制 这些效应需要使用能够检测特定神经回路中的生理变化的工具， 以及在行为分析期间调节它们的活动的能力。在此指导阶段（K99）， 在一项提议中，我将学习使用病毒遗传策略来可视化受酗酒影响的PFC预测， 使用离体方法表征青少年酗酒后兴奋性的回路特异性变化 电生理学此外，我将接受训练，利用化学遗传学调节行为过程中的PFC活性， 为了确定这些前额叶通路在酗酒和工作中的具体作用， 记忆在R00阶段，我将利用一个新开发的系统， 在定义的行为期间活跃的神经元集合（FosTRAP）。结合这些技术 在指导阶段学到的知识，我将使用这种技术来识别、表征和调节神经元 参加狂欢式饮酒的团体。总的来说，这个提议中的实验是 旨在测试总体假设，即青少年酗酒对内在的 内侧前额叶皮质锥体神经元亚群的兴奋性以及这些亚群发挥不同的作用 酗酒和工作记忆的影响所提议的实验将整合我以前的 行为药理学、免疫组织化学和电生理学培训， 识别和操纵神经回路的策略。该培训计划与提供的指导相结合， 我的导师，将大大提高我的能力，以回答有关的重要研究问题， 酒精中毒的神经生物学，同时促进我成为一名独立研究科学家。

PROJECT SUMMARY Excessive alcohol consumption has widespread personal and societal consequences, negatively affecting individual health while creating a significant economic and legal burden. Despite increasing efforts over the last few decades to identify genetic influences and neuroadaptations that are associated with the development of alcohol use disorders (AUDs), progress has been limited by the complexity of the underlying neuroanatomy. It is clear that the experimental resolution needs to be improved to the level of identifying adaptations in specific neural circuits that underlie dissociable behaviors related to AUD pathology. Thus, the goal of my research is to identify the neuroadaptations resulting from adolescent binge drinking that perpetuate heavy drinking and cognitive deficits in adulthood. I have focused my research on the prefrontal cortex (PFC), as this region continues to develop during adolescence and may be vulnerable to heavy alcohol consumption. For instance, PFC dysfunction is observed in binge drinkers and likely contributes to compulsive alcohol drinking and cognitive deficits observed in AUDs. In mice, I have found that binge drinking during adolescence disrupts performance on a PFC-dependent working memory task, increases alcohol consumption in adulthood, and significantly alters the intrinsic excitability of PFC pyramidal neurons. Discerning the mechanisms underlying these effects requires the use of tools capable of detecting physiological changes in specific neural circuits, as well as the ability to modulate their activity during behavioral analyses. In the mentored phase (K99) of this proposal, I will learn to use viral genetic strategies to visualize PFC projections affected by binge drinking and characterize the circuit-specific changes in excitability following adolescent binge drinking using ex vivo electrophysiology. Further, I will be trained to modulate PFC activity during behavior using chemogenetics, toward the goal of determining the specific role of these prefrontal pathways in binge-drinking and working memory. In the R00 phase, I will utilize a newly developed system for gaining permanent genetic access to neuronal ensembles that are active during defined behaviors (FosTRAP). In combination with the techniques learned in the mentored phase, I will use this technique to identify, characterize and modulate neuronal ensembles engaged in binge-like alcohol consumption. Taken together, the experiments in this proposal were designed to test the overarching hypothesis that adolescent binge drinking differentially affects the intrinsic excitability of medial PFC pyramidal neuron subpopulations and that these subpopulations play separate roles in binge alcohol consumption and working memory. The proposed experiments will integrate my previous training in behavioral pharmacology, immunohistochemistry and electrophysiology with new viral genetic strategies to identify and manipulate neural circuits. This training plan, in combination with guidance provided by my mentors, will greatly improve my ability to answer important research questions regarding the neurobiology of alcoholism while promoting my transition into an independent research scientist.