Prefrontal Pathways Engaged in Excessive Alcohol Consumption

项目总结 过度饮酒具有广泛的个人和社会后果，负面影响 个人健康，同时造成重大的经济和法律负担。尽管在过去的几年里不断努力 几十年来确定与糖尿病的发展相关的遗传影响和神经适应 在酒精使用障碍(AUDS)方面，由于潜在神经解剖学的复杂性，进展受到限制。它 很明显，实验分辨率需要提高到识别特定适应的水平 神经回路是与AUD病理相关的分离行为的基础。因此，我的研究目标是 为了确定青少年酗酒导致的神经适应，使大量饮酒和 成年后的认知缺陷。我把我的研究重点放在前额叶皮质(PFC)上，就像这个区域 在青春期继续发展，可能容易受到大量饮酒的影响。例如, 在酗酒者中观察到PFC功能障碍，并可能导致强迫性饮酒和 在AUDS中观察到认知缺陷。在老鼠身上，我发现青春期酗酒会扰乱 在依赖于PFC的工作记忆任务中的表现会增加成年后的酒精消耗量，并且 显著改变PFC锥体神经元的内在兴奋性。洞察潜在的机制 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，如 以及在行为分析期间调节它们活动的能力。在此的指导阶段(K99) 提案中，我将学习使用病毒遗传策略来可视化受酗酒和 用体外实验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PFC的活动， 旨在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，我将利用一种新开发的系统获得永久的遗传访问权限 在定义的行为中活跃的神经元集合(FosTRAP)。与这些技术相结合 在指导阶段学到的，我将使用这项技术来识别、表征和调节神经元 像狂欢一样酗酒的团体。综上所述，该提案中的实验是 旨在测试青少年酗酒会不同地影响人的内在机能这一重要假设 前额叶内侧锥体神经元亚群的兴奋性及其各自的作用 在酗酒和工作记忆方面。拟议的实验将整合我之前的 接受新病毒基因的行为药理学、免疫组织化学和电生理学方面的培训 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 将极大地提高我回答有关 酒精中毒的神经生物学，同时促进我向独立研究科学家的转变。

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.