Neuroplasticity of the Extended Amygdala CRF circuitry in alcohol dependence

酒精使用障碍(AUD)是一种主要的公共卫生问题，其特征是失去对 饮酒和戒酒期间出现的消极情绪。这个 过渡到酒精依赖与伴随的执行功能失调有关 通过内侧前额叶皮质(MPFC)和促肾上腺皮质激素释放因子(CRF)的募集 它的CRF受体1(CRF1)位于杏仁中央核(CEA)，但人们对此知之甚少 CRF/CRF1信号通路在皮质中的作用。至关重要的是，开发新的访问工具 CRF1神经元亚群为我们研究CRF/CRF1的功能和连接提供了便利 系统。在最初的拨款中，我们使用了一种创新的转基因小鼠模型，在该模型中，CRF1 共表达绿色荧光蛋白的(CRF1+)神经元(CRF1：GFP小鼠) 了解急性和慢性酒精对局部和下游CEA的细胞类型特异性影响 微电路。初步数据显示，CRF1+神经元在mPFC中高表达 2/3层，与未标记的相比，慢性酒精后经历了特定的神经适应 (CRF1-)神经元。因此，在这次更新中，我们将继续使用CRF1：GFP和CRF1：CRE小鼠来 CRF1+和CRF1+的电生理、神经化学和形态特征 并阐明了mPFC CRF1+通路在焦虑症和非焦虑症中的作用。 饮酒行为。通过系统生物学的方法，我们将描绘CRF1依赖的效应 酒精在mPFC中的表达，以及可能调节酒精依赖的杏仁核-mPFC连接 和退缩，并导致相关的焦虑相关行为。此外，我们还将使用 荧光激活细胞分选和全转录组分析揭示分子 这个特定的CRF1皮质群体的特征和神经适应，这可能识别新的 为酒精中毒的治疗策略提供了有希望的目标。

Alcohol use disorder (AUD) is a major public health concern characterized by the loss of control over alcohol drinking and the emergence of negative emotionality during abstinence from alcohol. The transition to alcohol dependence is associated with the concomitant dysregulation of executive function by the medial prefrontal cortex (mPFC) and the recruitment of corticotropin releasing factor (CRF) and its CRF receptor 1 (CRF1) in the central nucleus of the amygdala (CeA), but much less is known about the role of CRF/CRF1 signaling in cortical areas. Critically, the development of new tools to access subpopulations of CRF1 neurons has facilitated our studies of function and connectivity of CRF/CRF1 systems. In the original grant, we use an innovative transgenic mouse models in which CRF1 expressing (CRF1+) neurons co-express green fluorescent protein (GFP) (CRF1:GFP mice) to understand the cell type-specific effects of acute and chronic alcohol in local and downstream CeA microcircuits. Preliminary data have revealed that the CRF1+ neurons are highly expressed in mPFC layer 2/3 and undergo specific neuroadaptations following chronic alcohol compared to unlabeled (CRF1-) neurons. Thus, in this renewal, we will continue to use CRF1:GFP and CRF1:Cre mice to characterize the electrophysiological, neurochemical and morphological properties of both CRF1+ and CRF1- neurons in the mPFC, and elucidate the role of mPFC CRF1+ circuitry in anxiety-like and drinking behaviors. Through a systems biology approach, we will delineate the CRF1-dependent effects of ethanol in mPFC, as well as amygdala-mPFC connectivity that may regulate ethanol dependence and withdrawal and contribute to associated anxiety-related behaviors. In addition, we will use fluorescence activated cell sorting (FACS) followed by whole transcriptome analysis to reveal molecular signatures and neuroadaptations of this specific CRF1 cortical population, which may identify new promising targets for treatment strategies for alcoholism.