The role of intermittent binge ethanol, sex, and age on memory and CREB binding protein (CBP) expression

摘要 在人类中，青春期是一个关键的发育时期，以冒险行为的增加为标志，大脑 成熟和认知发展。对于大多数未成年人来说，酒精是一种很容易吸毒的东西 饮酒者要获得。早年饮酒会增加一个人晚年酗酒和依赖的风险 并可能导致蛋白质表达和行为的持久变化。青少年有一种不同的敏感度 酒精可能是由于它们正在发育中，对负面影响的敏感度降低，而 与成年人相比，对奖励效果的敏感度增加。这种发展上的差异 可能会进一步促进青少年饮酒，导致更严重的长期变化 而不是成年人。饮酒导致蛋白质表达的变化和记忆障碍 在成年人和青少年中都能观察到。两种已知的与记忆有关的蛋白质是cAMP反应 元素结合蛋白(CREB)和CREB结合蛋白(CBP)。CBP是一种组蛋白乙酰转移酶， 包括CREB在内的许多转录因子的辅活化子/辅因子，对短期和长期都是必需的- 术语记忆。海马体和前额叶皮质是经历成熟变化的两个大脑区域。 并在青春期起到记忆的作用。性也被认为在记忆中起着作用，研究表明 这表明女性可能对酒精的影响更敏感。考虑到酒精可能产生的持久影响 在行为和大脑方面，进一步了解酒精对性别和年龄的持续影响是 对文献和进一步治疗的可能性很重要。我们的实验室已经证明乙醇对 最后一次服药后3周的记忆，青少年在新物体识别任务中观察到的缺陷 处理的小鼠，而不是成年处理的小鼠。此外，我们还发现了几个与CRE、CREB和CBP相关的基因 在基因芯片分析后，由于乙醇的作用而减少。这项提案将研究性的影响， 酒精和发育年龄对记忆和CBP蛋白表达的行为和分子研究 在DBA/2J小鼠体内进行检测。目标1将研究酗酒对乙醇代谢的影响 乙醇对青春期和成年期雄性和雌性小鼠记忆和认知灵活性的影响 使用血液酒精浓度和巴恩斯迷宫任务。在目标2中，我将评估蛋白质水平 使用西语表达CBP和其他与CBP相互作用或参与Cre/CREB的蛋白质。我 还将使用免疫共沉淀和质谱学相结合的方法来评估CBP蛋白质的相互作用。 最后，在目标3中，我将使用病毒载体在青春期狂欢后的小鼠身上测试CBP的充分性 乙醇程序来评估对前面提到的记忆任务的影响。这些结合在一起 行为和分子研究将为解决以下问题的文献提供有用的知识 发育年龄、性别和酒精对记忆和CBP途径的影响。

Summary In humans, adolescence is a critical developmental period marked by increased risk-taking behaviors, brain maturation, and cognitive development. Alcohol is highly consumed as an easy drug for most underage drinkers to obtain. The early age of drinking increases one’s risk of alcohol abuse and dependence later in life and can lead to lasting changes in protein expression and behavior. Adolescents have an altered sensitivity to alcohol likely due to their ongoing development, showing decreased sensitivity to the negative effects while having increased sensitivity to the rewarding effects as compared to adults. This difference in development may further promote alcohol drinking in adolescents, resulting in long-lasting changes that are more severe than in adults. Alcohol consumption has resulted in changes in protein expression and memory impairments observed in both adults and adolescents. Two known proteins involved in memory are cAMP-response element binding (CREB) protein and CREB binding protein (CBP). CBP is a histone acetyltransferase, coactivator/cofactor for many transcription factors including CREB, and necessary for both short term and long- term memory. The hippocampus and prefrontal cortex are two brain regions that undergo maturation changes during adolescence and play a role in memory. Sex is also known to play a role in memory and studies have suggested women may be more sensitive to the effects of alcohol. Given the lasting impact alcohol can have on behavior and the brain, further understanding the persistent effects of alcohol due to sex and age is important for the literature and further treatment possibilities. Our lab has shown ethanol to differentially impact memory 3 weeks after the last dose, with an observed deficit in the novel object recognition task in adolescent treated mice but not adult treated mice. Furthermore, we found several genes related to CRE, CREB, and CBP were decreased due to ethanol following microarray analysis. This proposal will investigate the effects of sex, ethanol, and developmental age on memory and CBP protein expression using behavioral and molecular assays in DBA/2J mice. Aim 1 will investigate the impact of binge ethanol on ethanol metabolism, spatial memory, and cognitive flexibility in male and female mice treated with ethanol in adolescence and adulthood using blood ethanol concentration and the Barnes Maze task. In Aim 2, I will assess the level of protein expression of CBP and other proteins that interact with CBP or are involved with CRE/CREB using westerns. I will also assess CBP protein interactions using co-immunoprecipitation coupled with mass spectrometry. Finally, in Aim 3 I will test the sufficiency of CBP using viral vectors in mice following the adolescent binge ethanol procedure to assess the impact on the previously mentioned memory tasks. The combination of these behavioral and molecular studies will provide useful knowledge to the literature addressing the gap regarding developmental age, sex, and ethanol in memory and the CBP pathway.