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

总结 对于人类来说，青春期是一个关键的发育时期，其特点是冒险行为、大脑 成熟和认知发展。对于大多数未成年人来说，酒精是一种容易吸食的毒品，因此被大量消费 饮酒者获得。早期饮酒会增加晚年酗酒和依赖酒精的风险 并可以导致蛋白质表达和行为的持久变化。青少年的敏感度发生了改变 酒精可能是由于它们的持续发育，显示出对负面影响的敏感性降低，而 与成年人相比，他们对奖励效应的敏感性更高。这种发展上的差异 可能会进一步促进青少年饮酒，导致更严重的长期持续变化 比成人。饮酒导致蛋白质表达变化和记忆障碍 在成人和青少年中均观察到。两种已知的参与记忆的蛋白质是 cAMP 反应 元件结合（CREB）蛋白和CREB结合蛋白（CBP）。 CBP是一种组蛋白乙酰转移酶， 许多转录因子（包括 CREB）的辅激活因子/辅因子，对于短期和长期转录都是必需的。 术语记忆。海马体和前额叶皮层是经历成熟变化的两个大脑区域 在青春期并在记忆中发挥作用。众所周知，性对记忆也有影响，研究表明 建议女性可能对酒精的影响更敏感。鉴于酒精可能产生的持久影响 对行为和大脑的影响，进一步了解酒精对性别和年龄的持续影响是 对于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会产生不同的影响 最后一次给药后 3 周的记忆力，在青少年的新物体识别任务中观察到缺陷 治疗的小鼠，但不是成年治疗的小鼠。此外，我们还发现了几个与CRE、CREB和CBP相关的基因 微阵列分析后由于乙醇而减少。该提案将调查性的影响， 使用行为和分子方法研究乙醇和发育年龄对记忆和 CBP 蛋白表达的影响 DBA/2J 小鼠中的测定。目标 1 将研究酗酒对乙醇代谢、空间和空间的影响。 在青春期和成年期接受乙醇处理的雄性和雌性小鼠的记忆力和认知灵活性 使用血液乙醇浓度和巴恩斯迷宫任务。在目标 2 中，我将评估蛋白质水平 使用蛋白质印迹法表达 CBP 和其他与 CBP 相互作用或与 CRE/CREB 相关的蛋白质。我 还将使用免疫共沉淀结合质谱法评估 CBP 蛋白质相互作用。 最后，在目标 3 中，我将在青少年暴饮暴食后使用病毒载体在小鼠中测试 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。这些的组合 行为和分子研究将为文献提供有用的知识，解决以下方面的差距 发育年龄、性别和记忆中的乙醇以及 CBP 途径。

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.