Role of CAMKII in ethanol self-administration

描述（由申请人提供）：酒精成瘾是一种疾病，可以负面影响人们的关系，生产力，并可能导致重大的健康问题，包括死亡。据估计，1760万美国人患有酒精滥用或酒精中毒，但有效的治疗方法很少，这意味着需要更好地了解酒精如何影响哺乳动物的大脑。该提案的目标是更好地了解介导酒精消费和复发的分子机制。人们普遍认为，饮酒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。导致酒精寻求行为的大脑回路特别脆弱，它们的失调可能介导与酒精成瘾相关的行为病理。杏仁核是一个令人感兴趣的大脑区域，它调节酒精强化和奖励相关的学习。初步数据将钙/钙调蛋白激酶II（CAMKII）的α亚基鉴定为钙信号传导蛋白激酶，其在C57 BL/6 J小鼠中慢性酒精自我施用后在杏仁核中上调。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仍然未知。该项目试图检验以下总体假设：在自我饮酒后CAMKII增加，并且调节CAMKII活性在三个独立但综合的具体目标中功能性地调节寻求酒精的行为：1）使用蛋白质印迹和免疫组织化学分析来表征长期自我饮酒对CAMKII通路的影响; 2）检查操作性酒精自我给药对CAMKII活性的影响，并使用将药理学CAMKII抑制剂位点特异性输注到杏仁核中来确定CAMKII是否在功能上调节酒精在小鼠中的增强作用;和3）使用CAMKII抑制剂的位点特异性输注，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的恢复，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学特征。

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.