Molecular Component - Contet

摘要-分子组件 TSRI-ARC作为一个完整的整体，将专注于长期戒断的神经生物学及其在 复发和康复。分子成分将检验这样一种假设，即成分的变化 和/或微管(MT)细胞骨架的动力学有助于动机和情绪 通过驱动大脑关键区域神经元的结构重塑而产生的戒断症状学。 第一个具体目标将是使用最先进的技术来确定MT组成和动态的变化 蛋白质组学方法定量α和β-微管蛋白亚型的丰度，已知MT相关蛋白 调节MT的动力学，以及翻译后的微管蛋白修饰指示MT的稳定性。这些分析 将在内侧前额叶皮质(MPFC)、前岛和杏仁核进行。小白鼠将暴露在 慢性间歇性酒精吸入(CIE)和大脑将在戒断后1周和4周收集，以探讨 戒断早期和晚期的分子适应的发展和持续。第二 具体目的将是测试MT改变在酒精饮酒升级中的功能含义，阴性 戒断过程中的影响和树突重塑。第一种方法将涉及局部击倒微管蛋白 同种类型和MT相关蛋白，以模拟或逆转特定目标1中确定的丰度变化。 另一种方法将需要使用合成孕烯醇酮衍生物MAP4343进行全身治疗，以刺激 MT的动态和加速戒断的恢复。第三个具体目标将是补充研究 由其他TSRI-ARC组件执行。第一个实验将探索糖皮质激素的参与 CIE对MT动力学影响的受体信号转导。第二个实验将测试5-羟色胺能 与CIE戒断相关的焦虑样行为中杏仁中央核和mPFC的投射。 综上所述，该项目有望确定内环境平衡失败的分子机制--或 异位--这是酒精使用障碍的特征，也是长期酗酒易复发的原因 禁欲。分子成分将与动物模型核心有很强的相互作用，将涉及 与神经生理学和神经回路组件合作，并将提供机械基础 用于由临床组件对化合物进行测试。

SUMMARY – Molecular Component The TSRI-ARC as an integrated whole will focus on the neurobiology of protracted abstinence and its role in relapse and recovery. The Molecular Component will test the hypothesis that alterations in the composition and/or dynamics of the microtubule (MT) cytoskeleton contribute to the motivational and emotional symptomatology of abstinence by driving the structural remodeling of neurons in key brain regions. The first Specific Aim will be to determine alterations in MT composition and dynamics, using state-of-the-art proteomics methodology to quantify the abundance of α- and β-tubulin isotypes, MT-associated proteins known to regulate MT dynamics, and tubulin posttranslational modifications indicative of MT stability. These analyses will be performed in the medial prefrontal cortex (mPFC), anterior insula, and amygdala. Mice will be exposed to chronic intermittent ethanol inhalation (CIE) and brains will be collected 1 and 4 weeks into withdrawal to explore the development and persistence of molecular adaptations in early and late abstinence, respectively. The second Specific Aim will be to test the functional implication of MT alterations in ethanol drinking escalation, negative affect and dendritic remodeling during abstinence. A first approach will involve local knockdown of tubulin isotypes and MT-associated proteins to mimic or reverse abundance changes identified in Specific Aim 1. Another approach will entail systemic treatment with the synthetic pregnenolone derivative MAP4343 to stimulate MT dynamics and accelerate recovery from withdrawal. The third Specific Aim will be to complement research conducted by other TSRI-ARC components. A first experiment will probe the involvement of glucocorticoid receptor signaling in the effect of CIE on MT dynamics. A second experiment will test the role of serotonergic projections to the central amygdala and mPFC in the anxiety-like behavior associated with abstinence from CIE. Altogether, this project is anticipated to identify a molecular mechanism for the homeostatic failure—or allostasis—that characterizes alcohol use disorders and underlies relapse vulnerability during protracted abstinence. The Molecular Component will have strong interactions with the Animal Models Core, will involve collaborations with the Neurophysiology and Neurocircuitry Components, and will provide a mechanistic basis for the testing of compounds by the Clinical Component.