Neuroplasticity of the Extended Amygdala CRF circuitry in alcohol dependence

描述（由申请人提供）：最近的研究强调了杏仁核中央核（CeA）和促肾上腺皮质激素释放因子（CRF）系统在乙醇依赖性发展中的作用。特别是，CRF 1型受体（CRF1）拮抗剂阻断焦虑的影响和增加乙醇自我管理所产生的压力和乙醇戒断。值得注意的是，慢性CRF1拮抗作用消除了慢性间歇性乙醇暴露大鼠中乙醇饮用的依赖性诱导的升级。我们证明，急性乙醇和CRF通过CRF1激活增加小鼠和大鼠CeA中的GABA能传递，并且CRF和CRF1拮抗剂对乙醇依赖性大鼠CeA神经元中GABA能传递的影响增强。然而，CeA由异质细胞类型组成，并且技术限制已经排除了对从中获得记录的神经元的鉴定和进一步表征。因此，尽管积累了大量的知识，急性和慢性乙醇在这个大脑区域的CRF1介导的影响，潜在的局部神经元微电路需要划定。 因此，在这项研究中，我们将使用一种创新的转基因小鼠模型，表达绿色荧光蛋白（GFP）在表达CRF1的神经元（CRF1：GFP小鼠）比较CeA CRF1表达神经元的电生理，神经化学，形态学和hodological特性的未标记的神经元。使用电生理学与组织化学和神经示踪方法相结合，我们将获得新的信息，这些神经元的功能和连接，从而提供机制的CRF系统在酒精依赖的发展中的作用的见解。Contet博士和Mandyam博士是参与本研究的关键人员，拥有完成这样一个多学科项目所需的所有必要专业知识。 更好地了解乙醇效应的分子机制和乙醇依赖性中CRF神经回路的神经适应性对酒精研究人员来说是一个挑战，并将有助于发现新的治疗药物来缓解酒精中毒。

DESCRIPTION (provided by applicant): Recent research has highlighted the role of the central nucleus of the amygdala (CeA) and the recruitment of the corticotropin-releasing factor (CRF) system in the development of ethanol dependence. Particularly, CRF type 1 receptor (CRF1) antagonists block the anxiogenic effects and the increase in ethanol self-administration produced by stressors and ethanol withdrawal. Notably, chronic CRF1 antagonism abolishes dependence-induced escalation of ethanol drinking in rats exposed to chronic intermittent ethanol. We demonstrated that acute ethanol and CRF increase GABAergic transmission in mouse and rat CeA via CRF1 activation, and the effects of CRF and CRF1 antagonists on GABAergic transmission are enhanced in CeA neurons from ethanol-dependent rats. However, the CeA is comprised of heterogeneous cell types and technical limitations have precluded the identification and further characterization of neurons from which recordings were obtained. Thus, despite the vast knowledge accumulated on the CRF1-mediated effects of acute and chronic ethanol in this brain region, the underlying local neuronal microcircuitry needs to be delineated. Thus, in this research proposal we will use an innovative transgenic mouse model expressing green fluorescent protein (GFP) in neurons expressing CRF1 (CRF1:GFP mice) to compare the electrophysiological, neurochemical, morphological and hodological properties of CeA CRF1-expressing neurons to those of unlabeled neurons. Using electrophysiology in combination with histochemical and neurotracing methods, we will obtain novel information on functionality and connectivity of these neurons, thereby providing mechanistic insights into the role of the CRF system in the development of alcohol dependence. Drs. Contet and Mandyam, key personnel involved in the present study, possess all the necessary expertise needed to accomplish such a multidisciplinary project. A better understanding of the molecular mechanisms underlying ethanol effects and the neuroadaptations shaping the CRF neurocircuitry involved in ethanol-dependence represent a challenge to alcohol researchers and will be useful toward uncovering new therapeutic agents to alleviate alcoholism.