Role of CAMKII in ethanol self-administration

描述（由申请人提供）：酒精成瘾是一种疾病，会对人们的人际关系、生产力产生负面影响，并可能导致包括死亡在内的重大健康问题。据估计，1760万美国人患有酒精滥用或酒精中毒，但有效的治疗方法却很少，这表明有必要更好地了解酒精如何影响哺乳动物的大脑。这项提议的目的是为了更好地理解介导酒精消耗和复发的分子机制。人们普遍认为，饮酒会导致调节神经可塑性的细胞内信号级联的神经适应性变化。导致寻求酒精行为的大脑回路特别脆弱，它们的失调可能会介导与酒精成瘾相关的行为病理。我们感兴趣的一个大脑区域是杏仁核，它调节酒精强化和与奖励相关的学习。初步数据表明，C57BL/6J小鼠慢性酒精自我给药后，钙/钙调蛋白激酶II （CAMKII）的α亚基是一种钙信号蛋白激酶，在杏仁核中上调。CAMKII调节受体活性并启动控制神经可塑性的多种转录因子，但CAMKII在酒精自我给药中的功能参与尚不清楚。本项目旨在测试CAMKII在自我饮酒后增加的总体假设，以及调节CAMKII活性在三个独立但综合的特定目标中功能性地调节寻酒行为：1)使用western blot和免疫组织化学分析表征慢性自我饮酒对CAMKII途径的影响；2)通过特定部位向杏仁核内注射CAMKII抑制剂，研究操作性酒精自我给药对CAMKII活性的影响，并确定CAMKII是否在功能上调节酒精对小鼠的强化作用；3)通过特定部位注射CAMKII抑制剂，确定杏仁核中的CAMKII是否在功能上调节线索诱导的酒精寻求恢复，这是一种复发的行为模型。这些临床前实验有可能揭示一种新的分子机制，这种机制是酒精强化和复发的基础，是酒精成瘾的行为病理特征。

DESCRIPTION (provided by applicant): Alcohol addiction is a disease that can negatively affect people's relationships, productivity and can lead to major health problems including death. It is estimated that 17.6 million Americans suffer from alcohol abuse or alcoholism yet there are very few effective treatments available signifying the need to better understand how alcohol affects the mammalian brain. This goal of this proposal is to better understand the molecular mechanisms that mediate alcohol consumption and relapse. It is widely accepted that alcohol consumption leads to neuroadaptive changes in intracellular signaling cascades that regulate neuroplasticity. Brain circuits that contribute to alcohol seeking behaviors are particularly vulnerable and their dysregulation may mediate behavior pathologies associated with alcohol addiction. One brain region of interest is the amygdala, which regulates alcohol reinforcement and reward- related learning. Preliminary data identified the alpha subunit of calcium/calmodulin kinase II (CAMKII) as a calcium-signaling protein kinase that is upregulated in the amygdala following chronic alcohol self- administration in C57BL/6J mice. CAMKII modulates receptor activity and initiates multiple transcription factors that control neural plasticity, yet the functional involvement of CAMKII in alcohol self-administration remains unknown. This project seeks to test the overall hypothesis that CAMKII is increased following alcohol self-administration and that modulating CAMKII activity functionally regulates alcohol-seeking behaviors in three separate but integrated specific aims: 1) to characterize the effect of chronic alcohol self- administration on the CAMKII pathway using western blot and immunohistochemical analysis; 2) to examine the effect of operant alcohol self-administration on CAMKII activity and determine if CAMKII functionally regulates the reinforcing effects of alcohol in mice using site-specific infusions of a pharmacological CAMKII inhibitor into the amygdala; and 3) to determine if CAMKII in the amygdala functionally regulates cue- induced reinstatement of alcohol-seeking, a behavioral model of relapse, using site-specific infusions of the CAMKII inhibitor. These preclinical experiments have the potential to reveal a novel molecular mechanism that underlies alcohol reinforcement and relapse, behavioral pathologies that characterize alcohol addiction.