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

总结 在人类中，青春期是一个关键的发展时期，其特征是冒险行为增加，大脑 成熟和认知发展。酒精作为一种容易吸食的毒品，对大多数未成年人来说， 酒客们来拿。过早饮酒会增加一个人日后酗酒和依赖酒精的风险 并可导致蛋白质表达和行为的持久变化。青少年对 酒精可能是由于他们正在进行的发展，显示出对负面影响的敏感性下降， 与成年人相比，对奖励效应的敏感性增加。这种发展的差异 可能会进一步促进青少年饮酒，导致更严重的长期变化， 比成年人的多。饮酒导致蛋白质表达的变化和记忆障碍 在成人和青少年中观察到。两种已知的与记忆有关的蛋白质是cAMP反应蛋白 元件结合蛋白（CREB）和CREB结合蛋白（CBP）。CBP是组蛋白乙酰转移酶， 它是许多转录因子（包括CREB）的辅激活因子/辅因子，也是短期和长期转录所必需的。 术语记忆海马体和前额叶皮层是两个经历成熟变化的大脑区域 并在记忆中发挥作用。众所周知，性在记忆中也起着一定的作用， 女性可能对酒精的影响更敏感。考虑到酒精的持久影响 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， 重要的文献和进一步的治疗可能性。我们的实验室已经证明乙醇对 末次给药后3周的记忆，在青少年中观察到新物体识别任务缺陷 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 在微阵列分析后由于乙醇而降低。这项提案将调查性的影响， 乙醇和发育年龄对记忆和CBP蛋白表达的影响 在DBA/2 J小鼠中的测定。目的1将研究酗酒对乙醇代谢的影响， 在青春期和成年期用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 CBP和其他与CBP相互作用或与CRE/CREB相关的蛋白质的表达。我 还将使用免疫共沉淀结合质谱法评估CBP蛋白相互作用。 最后，在目标3中，我将在青少年狂欢后使用病毒载体在小鼠中测试CBP的充分性 乙醇程序，以评估对先前提到的记忆任务的影响。结合这些 行为学和分子学研究将为解决以下差距的文献提供有用的知识： 发育年龄，性别，乙醇在记忆和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.