Project 3: Adolescent vulnerability to chronic ethanol: neurophysiological, biochemical, and behavioral mechanisms of adult AUD

总结 控制从酒精使用过渡到滥用的神经生物学机制知之甚少。 我们的总体方法是检查脆弱人群，以突出特定的细胞/分子途径 参与到这一转变中。例如，暴露于大量饮酒的人类青少年的死亡率要高得多。 成年后酗酒的风险。最近的研究表明，青少年也有类似的责任。 包括啮齿动物在内的动物。我们发表的研究表明，青少年慢性乙醇暴露差异 调节外侧/基底外侧杏仁核（BLA）中的谷氨酸能和GABA能神经传递， 在情绪反应期间，对整合认知和感觉信息至关重要的回路内的“节点”， 以特定于输入的方式。我们的数据还表明哺乳动物雷帕霉素靶蛋白（mTOR）的关键作用- 在突触强化中依赖信号级联。此外，我们提供的初步数据表明， 这些突触效应中的许多在成年动物中不存在或大大减弱。这表明mTOR- 依赖性信号直接调节青少年乙醇暴露期间的突触调制。整体 因此，本项目的目标是使用成熟的青春期大鼠乙醇蒸汽暴露， 通过整合细胞、分子和生物学信息，了解这种暴露对成年动物的长期影响。 行为方法论拟议的工作包括三个具体目标：目标1将描述影响 青少年乙醇暴露对成人BLA能和GABA能神经传递的影响 细胞膜片钳电生理学;目的2将描述青少年乙醇暴露对mTOR的影响 BLA中突触后和突触前区室的信号传导;目标3将检查 药物干预沿着mTOR信号通路及其对长期行为的影响 青少年乙醇暴露的后果。这些目标合在一起意义重大，因为它们利用了 脆弱群体（青少年）、创新的技术和概念方法， 我们的研究团队的专业知识，以帮助确定特定的细胞信号传导过程， 乙醇暴露在多个水平的分析。我们将直接测试这些信号过程是否代表 潜在的治疗目标，旨在中断从乙醇使用过渡到滥用。

SUMMARY The neurobiological mechanisms controlling the transition from alcohol use to abuse are poorly understood. Our overall approach is to examine vulnerable populations to highlight specific cellular/molecular pathways involved in this transition. For example, human adolescents exposed to heavy alcohol use are at much greater risk for the development of alcoholism as adults. Recent studies suggest similar liabilities for adolescent animals including rodents. Our published work indicates adolescent chronic ethanol exposure differentially modulates both glutamatergic and GABAergic neurotransmission in the lateral/basolateral amygdala (BLA), a `node' within circuits critical for the integration cognitive and sensory information during emotional responses, in an input-specific fashion. Our data also suggest a critical role for mammalian target of rapamycin (mTOR)- dependent signaling cascades in synaptic strengthening. Further, we provide preliminary data suggesting that many of these synaptic effects are absent or greatly diminished in adult animals. This suggests that mTOR- dependent signaling directly regulates synaptic modulation during adolescent ethanol exposure. The overall goal of the current project is to therefore use a well-established ethanol vapor exposure in adolescent rats to understand the long-term impact of this exposure in adult animals by integrating cellular, molecular, and behavioral methodologies. The proposed work includes three specific aims: Aim 1 will characterize the effects of adolescent ethanol exposure on adult BLA glutamatergic and GABAergic neurotransmission using whole- cell patch clamp electrophysiology; Aim 2 will describe the effect of adolescent ethanol exposure on mTOR signaling in both postsynaptic and presynaptic compartments in the BLA; and, Aim 3 will examine pharmacological intervention along the mTOR-signaling pathway and its impact on the long-term behavioral consequences of adolescent ethanol exposure. Together these aims are significant because they leverage a vulnerable population (adolescents), innovative technical and conceptual approaches, and the substantial expertise of our research team to help identify specific cellular signaling processes governing the impact of ethanol exposure across multiple levels of analysis. We will directly test if these signaling processes represent potential therapeutic targets for treatments designed to interrupt the transition from ethanol use to abuse.