Assessing the adolescent PFC proteome for mechanisms of escalated alcohol self-administration in adulthood

 描述(由申请人提供)：青春期是一个关键时期，在此期间饮酒会增加患终生酒精使用障碍(AUDS)的风险。然而，调节这种风险增加的神经生物学机制尚未完全确定。为了在理解青少年对AUDS的易感性方面开辟新的天地，我们建议对小鼠的前额叶皮质(PFC)蛋白质组进行高通量筛选，以确定青少年和成年小鼠大脑中被乙醇修饰的蛋白质相互作用网络。这些基于发现的实验将在青少年PFC中识别新的酒精敏感蛋白质网络。初步的蛋白质组学研究结果表明，大麻素受体相互作用蛋白1a(CRIP1a)在青少年PFC中是一种发育调节的酒精敏感蛋白。有证据表明，内源性大麻素系统在功能上调节啮齿动物的饮酒，大麻素受体的表达在发育中的大脑中发生了显著的变化。因此，这一应用中的实验还将确定CRIP1a在青少年和成人饮酒中的功能作用，并确定CRIP1a的表达是否介导青少年饮酒的长期后果。为了确定CRIP1a是否介导了青少年饮酒后滥用的长期风险，我们将使用病毒载体介导的CRIP1a在青少年和成年PFC中的过度表达来推动CRIP1a表达的长期增加，并测量这两个年龄组的成年后有效的酒精自我给药。由于这种操作将与酒精对青少年PFC的影响相反，我们预计它将保护青少年而不是成人治疗组免受酒精强化效应的影响。总之，这些实验将在青少年PFC中识别新的酒精敏感蛋白质网络，并确定这些网络中的特定节点CRIP1a是否在青春期酒精暴露后的成年期调节酒精自我给药升级的风险。

 DESCRIPTION (provided by applicant): Adolescence is a critical period during which alcohol drinking increases the risk of lifetime alcohol use disorders (AUDs). However, the neurobiological mechanisms that mediate this increased risk have not been fully identified. To break new ground in our understanding of adolescent vulnerability to AUDs, we propose to conduct a high-throughput screen of the prefrontal cortex (PFC) proteome in mice to identify protein interaction networks that are modified by ethanol in the adolescent and adult mouse brain. These discovery-based experiments will identify novel alcohol-sensitive protein networks in adolescent PFC. Preliminary proteomic findings identified the cannabinoid receptor interacting protein 1a (CRIP1a) as a developmentally regulated ethanol-sensitive protein in the adolescent PFC. Evidence suggests that the endocannabinoid system functionally regulates alcohol drinking in rodents, and significant changes in expression of the cannabinoid receptors occur in the developing brain. Therefore, experiments in this application will also establish a functional role for CRIP1a in adolescent and adult alcohol drinking and determine whether CRIP1a expression mediates the long-term consequences of adolescent alcohol drinking. To determine whether CRIP1a mediates the long-term risk for abuse following adolescent alcohol drinking, we will use viral vector-mediated overexpression of CRIP1a in the adolescent and adult PFC to drive long-term increases in CRIP1a expression and measure operant self-administration of alcohol during adulthood in both age groups. As this manipulation will oppose the effect of ethanol in the adolescent PFC, we expect that it will be protective against enhanced reinforcing effects of alcohol in the adolescent but not adult treatment groups. Together, these experiments will identify novel alcohol sensitive protein-networks in the adolescent PFC and determine if a specific node in these networks, CRIP1a, mediates risk for escalated alcohol self-administration during adulthood after adolescent alcohol exposure.