Mechanisms and relevance of the ethanol-induced suppression of inhibitory signaling in the basolateral amygdala

项目摘要 一种新的共识是，上瘾是一种适应不良的学习方式，而接触毒品 例如乙醇(Etoh)通过改变学习的结构和/或功能来促进持续的药物使用- 相关的大脑区域。这一建议的前提是分子和细胞的鉴定 乙醇改变联想学习的机制将澄清乙醇成瘾的关键方面，并可能 导致改善酒精中毒的治疗方法。最近的工作，包括来自 这位候选人提出，乙醇可能通过影响G蛋白门控来调节联想学习 内向整流K+(GIRK)通道。初步数据显示，GIRK通道以 杏仁核基外侧核(BLA)是联想学习的关键结构，在大脑中反复出现。 活体乙醇暴露抑制该区域锥体/投射神经元中依赖于GIRK的信号。这 该提案将结合一系列跨学科的方法来测试乙醇诱导的 BLA锥体神经元GIRK通道活性的抑制有助于重复刺激的复杂效应 乙醇暴露，包括奖励信号和行为的变化。在目标1中，候选人将使用SLICE 电生理学方法探讨乙醇诱发的电生理特征及其机制 抑制GIRK信号。在AIM 2中，候选人将结合病毒介导的GIRK抑制和 偏好和奖励的行为测量，以及切片电生理和快速扫描周期 伏安法(FSCV)，评估BLA中GIRK抑制对奖赏相关的影响 神经传递和行为。该项目将提供高级研究培训的机会，将 协助应聘者未来的努力，包括测量兴奋性神经传递的方法 和脑片的可塑性，神经元对基因表达的特定操作，通过 快速扫描循环伏安法(FSCV)，并在小鼠中使用自愿的乙醇消耗模型。导师制 培训将由Kevin Wickman博士和Anna Lee博士提供。对该项目的其他支持以及 候选人的专业发展将来自Jeffery Weiner博士(顾问)，他是 乙醇对杏仁核生理的影响，以及由成瘾专家组成的当地咨询委员会 具有培训独立调查人员的良好记录的研究(Marilyn Carroll博士、Stan Thayer博士、 和罗伯特·梅塞尔)。这项提案的所有方面都是为了帮助候选人实现她的目标 过渡到一个独立的研究职位，在那里她打算制定一个研究计划， 调查Etoh在参与学习的地区的互连网络中的影响 支持上瘾的过程。

Project Summary There is an emerging consensus that addiction is a form of maladaptive learning, wherein exposure to drugs such as ethanol (EtOH) promotes continued drug use by altering the structure and/or function of learning- related brain regions. The premise of this proposal is that the identification of molecular and cellular mechanisms by which EtOH alters associative learning will clarify critical aspects of EtOH addiction and may lead to improved therapeutic approaches for the treatment of alcoholism. Recent work, including studies from the candidate, has suggested that EtOH may modulate associative learning via its effect on G protein-gated inwardly rectifying K+ (GIRK) channels. Preliminary data show that GIRK channels are expressed in the basolateral nucleus of the amygdala (BLA), a critical structure for associative learning, and that repeated in vivo EtOH exposure suppresses GIRK-dependent signaling in pyramidal/projection neurons in this region. This proposal will combine an array of interdisciplinary approaches to test the hypothesis that the EtOH-induced suppression of GIRK channel activity in BLA pyramidal neurons contributes to the complex effects of repeated EtOH exposure, including changes in reward signaling and behavior. In AIM 1, the candidate will utilize slice electrophysiology to probe the salient characteristics and underlying mechanisms of the EtOH-induced suppression of GIRK signaling. In AIM 2, the candidate will combine viral-mediated GIRK suppression with behavioral measures of preference and reward, as well as slice electrophysiology and fast scan cyclic voltammetry (FSCV), to assess the impact of GIRK suppression in the BLA on reward-related neurotransmission and behavior. This project will offer the opportunity for advanced research training that will assist the candidate in her future endeavors, including approaches to measuring excitatory neurotransmission and plasticity in brain slices, neuron-specific manipulation of gene expression, measuring dopamine release by fast-scan cyclic voltammetry (FSCV), and the use of voluntary EtOH consumption models in mice. Mentorship and training will be provided by Dr. Kevin Wickman and Dr. Anna Lee. Additional support for the project and the professional development of the candidate will come from Dr. Jeffery Weiner (Consultant), an expert in the effects of EtOH on amygdala physiology, and a local Advisory Committee composed of experts in addiction research with a strong track record of training independent investigators (Drs. Marilyn Carroll, Stan Thayer, and Robert Meisel). All facets of this proposal are designed to assist the candidate in achieving her goal of transitioning to an independent research position, where she intends to develop a research program that investigates the impact of EtOH across an interconnected network of regions involved in the learning processes that support addiction.