Chronic ethanol effects on cholinergic interneurons of the striatum

慢性乙醇对纹状体胆碱能中间神经元的影响 项目概述：纹状体与学习、奖励和成瘾有关，包括酗酒。在 在纹状体，消融胆碱能中间神经元(CINs)会导致类似于 在酗酒者身上观察到的那些。有趣的是，有证据表明慢性阻塞性肺疾病后CIN的减少。 酒精暴露。一致地，我发现以下猴子的纹状体胆碱能功能受损 长期的乙醇消费。因此，这一提议的首要假设是慢性 酒精暴露会降低纹状体CIN的数量或功能，并且这些酒精诱导的胆碱能 损伤是与酒精中毒相关的行为和认知缺陷的基础。为了测试这一点 假设，我提出了三个具体目标：具体目标1-I将确定急性乙醇对纹状体的影响 用转基因小鼠、免疫组织化学、RNAScope和电生理技术检测CIN亚型。 CIN亚型对酒精的敏感性特征将引起酒精和基底节的广泛关注。 研究人员都一样。特定目标2-I将确定慢性酒精诱导的纹状体CINs缺陷是否由于 CIN丧失，CIN功能受损，或两者兼而有之。文献中一致认为，纹状体 慢性酒精暴露后胆碱能环路功能低下，但这种功能障碍的根源并不是 安全。因此，我将确定CIN是否丢失和/或CIN功能是否受损(通过电生理学) 在长期接触酒精之后。这些结果将是第一次检验慢性乙醇对 纹状体CIN功能。然后，将确定CIN活性的体内化学发生操作 在慢性酒精中毒后，酒精足以改善伴随慢性酒精中毒的行为和认知障碍 酒精暴露包括酒精消耗量和在操作符颠倒学习任务中的表现。 在我的整个职业生涯中，我一直对正常和异常行为背后的机制感兴趣。我 我很幸运有导师教我从事有意义的研究所需的技能。在… 在我职业生涯的每个阶段，我的技术能力和科学成熟度都有所提高，随着我的继续 为了训练，我的导师金·布莱克威尔和大卫·洛文格将挑战我，让我作为一名科学家不断提高 提出有影响力的问题，设计令人信服的实验来解决这些问题，并提出我的 在手稿和演示文稿中清楚而有效地得出结论。此外，我们有一个计划，以确保我 接受培训，进行激动人心的实验，成功地运行实验室，并指导受训人员。此外，Dr. 阿德龙·哈里斯和玛丽莎·罗伯托这两位成功的研究人员已同意在我的顾问委员会任职 以确保我成功地过渡到一名独立调查员。因此，我相信，有了 我的导师、我的校外咨询委员会、我的技术顾问和机构 在NIAAA和乔治梅森大学的支持下，我将能够执行拟议的实验，获得 教师地位，并作为一名成功的独立神经科学家茁壮成长。 在这笔赠款的延期期间，我将继续为既定的具体目标而努力，我将能够 在R00阶段最初打算完成的一些目标上进行更多工作。尤其是，我会 能够进行目标3中指定的初步化学发生实验。我还将继续寻找 申请一个独立的教员职位。

Chronic ethanol effects on cholinergic interneurons of the striatum Project Summary: The striatum is implicated in learning, reward and addiction, including alcoholism. Within the striatum, ablation of cholinergic interneurons (CINs) results in behavioral and cognitive deficits similar to those observed in alcoholics. Interestingly, there is evidence indicating a decrease in CINs following chronic ethanol exposure. In agreement, I have found that striatal cholinergic function is impaired in monkeys following long-term ethanol consumption. Therefore, the overarching hypothesis of this proposal is that chronic ethanol exposure decreases striatal CIN numbers or function and that these ethanol-induced cholinergic impairments underlie the behavioral and cognitive deficits associated with alcoholism. To test this hypothesis, I propose three specific aims: Specific Aim 1 – I will determine the effect of acute ethanol on striatal CIN subtypes using transgenic mice, immunohistochemical, RNAscope, and electrophysiological techniques. The characterization of CIN subtype sensitivity to ethanol will be of broad interest to alcohol and basal ganglia researchers alike. Specific Aim 2 – I will determine if chronic ethanol-induced deficits in striatal CINs are due to a loss of CINs, an impairment of CIN function, or both. There is agreement in the literature that the striatal cholinergic circuit is hypo-functional following chronic ethanol exposure but the source of this dysfunction is not clear. Therefore, I will determine if CINs are lost and/or if CIN function is compromised (with electrophysiology) following chronic ethanol exposure. These results will be the first to examine the effects of chronic ethanol on striatal CIN function. Specific Aim 3 – I will then determine if in vivo chemogenetic manipulation of CIN activity after chronic ethanol are sufficient to ameliorate the behavioral and cognitive deficits that accompany chronic ethanol exposure including ethanol consumption and performance on an operant reversal learning task. Throughout my career, I have been interested in the mechanisms underlying normal and aberrant behaviors. I have been fortunate to have mentors that taught me the skills required to engage in meaningful research. At every stage of my career, I have advanced in my technical ability and scientific sophistication and as I continue to train, my mentors, Kim Blackwell and David Lovinger, will challenge me to constantly improve as a scientist and to ask impactful questions, to design compelling experiments to address those questions, and to present my findings clearly and effectively in manuscripts and presentations. Furthermore, we have a plan to ensure that I receive training to conduct exciting experiments, run a successful lab, and mentor trainees. In addition, Drs. Adron Harris and Marisa Roberto, two successful researchers, have agreed to serve on my advisory committee to ensure that I am successful in my transition to an independent investigator. Thus, I am confident that with the mentorship of my mentors, my extramural advisory committee, my technical consultants, and the institutional support of NIAAA and George Mason University, I will be able to execute the proposed experiments, attain a faculty position, and thrive as a successful independent neuroscientist. In the extension period of this grant, I will continue to work on the established specific aims and I will be able to conduct more work on some of the aims that were initially intended to done in the R00 phase. In particular, I will be able to conduct the preliminary chemogenetic experiments specified in aim 3. I will also continue to search for an independent faculty position.