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 纹状体与学习、奖励和成瘾有关，包括酗酒。内 在纹状体中，胆碱能中间神经元（CINs）的消融导致类似于 在酗酒者中观察到的。有趣的是，有证据表明慢性炎症后的CIN减少， 酒精暴露与此一致，我发现，在以下情况下，猴子纹状体胆碱能功能受损 长期乙醇消费。因此，这一提议的首要假设是， 乙醇暴露降低了纹状体CIN的数量或功能， 损害是与酗酒有关的行为和认知缺陷的基础。为了验证这一 假设，我提出了三个具体目标：具体目标1 -我将确定急性乙醇对纹状体的影响， 使用转基因小鼠、免疫组织化学、RNA显微镜和电生理技术进行CIN亚型分析。 CIN亚型对乙醇敏感性的特征将对乙醇和基底神经节产生广泛的兴趣 研究人员一样。具体目标2 -我将确定是否慢性乙醇诱导的缺陷，在纹状体CIN是由于 CIN丧失、CIN功能受损或两者。文献中一致认为， 慢性乙醇暴露后胆碱能回路功能减退，但这种功能障碍的来源不是 清楚因此，我将确定CIN是否丢失和/或CIN功能是否受损（电生理学） 慢性酒精暴露后。这些结果将是第一个研究慢性乙醇对 纹状体CIN功能。特异性目标3 - I然后将确定CIN活性的体内化学发生学操作是否 慢性酒精后足以改善伴随慢性酒精的行为和认知缺陷， 乙醇暴露，包括乙醇消耗和操作性逆转学习任务的表现。 在我的职业生涯中，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 在我职业生涯的每一个阶段，我都在提高我的技术能力和科学素养， 为了训练，我的导师金·布莱克威尔和大卫·洛文杰将挑战我作为一名科学家不断进步 并提出有影响力的问题，设计引人注目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晰有效地呈现调查结果。此外，我们有一个计划，以确保我 接受培训，进行令人兴奋的实验，运行一个成功的实验室，并指导学员。此外，博士。 阿德伦·哈里斯和玛丽莎·罗伯托，两位成功的研究人员，已经同意在我的顾问委员会任职。 以确保我能顺利过渡为独立调查员因此，我相信， 我的导师，我的校外咨询委员会，我的技术顾问和机构的指导 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 教师的位置，并茁壮成长为一个成功的独立神经科学家。 在此补助金的延长期内，我将继续致力于既定的具体目标，我将能够 就最初打算在R 00阶段完成的一些目标开展更多的工作。特别是，我将 能够进行目标3中规定的初步化学发生实验。我也会继续寻找 一个独立教师的职位

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.