CRF neurons of the extended amygdala and alcohol drinking

项目总结 酒精使用障碍是一个主要的公共健康问题。到目前为止，可用的治疗方案仅限于 心理社会干预，FDA批准的三种药物(双硫仑、氨基己酸酯和纳曲酮)和一种 很少有药物被批准用于其他适应症，所有这些药物的效果都相对较小。显然有必要 基于对神经生物学机制的更深入了解而进行的其他治疗。促肾上腺皮质激素 释放因子(CRF)是由41个氨基酸组成的神经肽，主要由室旁神经元产生。 下丘脑、中央杏仁核(CEA)和终纹床核(BNST)在其中起作用 在应激的行为和生理反应中起重要作用。长期以来，慢性肾功能衰竭一直与驾驶有关 在啮齿动物中使用CRF受体拮抗剂的先前研究表明，过量的酒精摄入。然而， 最近试图测试CRF受体拮抗剂作为人类酒精渴求的治疗方法 令人失望。这种不成功的部分原因可能是某些化合物的类药物特性不充分。 以及需要不同的人体实验室模型来测试药物对戒断和阴性的影响 依赖的受试者的强化。另一个原因可能是由于CRF从 神经元与其他多肽和神经递质可能协同作用，推动过度饮酒。 理解这些共同释放的因素中哪些是重要的，需要一个不同的战略，专注于 CRF神经元本身，而不是CRF受体。 缺乏CRF神经元亚群的遗传途径使CRF的生物学研究变得困难 神经元，CRF在不同大脑区域的来源，以及CRF调节行为的电路。 为了填补这一空白，我们建立了一个BAC转基因Wistar大鼠系，在其中Cre重组酶从 CRH基因启动子使CRF神经元能够获得遗传信息。在这个项目中，我们将利用这些老鼠来追踪 假设随着动物形成酒精依赖，CEA和BNST中的CRF神经元促进 通过GABA、CRF和其他神经肽的协调释放来消耗酒精。我们会 通过有选择地激活或抑制这些神经元群体及其投射来检验这一假设 使用化学遗传工具，我们将解决不同传递体和 通过依赖Cre的RNA干扰，这些神经元释放出调节剂。最后，我们将调查 反复酒精摄入对这些CRF神经元群体转录组的影响 了解乙醇如何改变它们的表型，这应该会为我们提供重要的新线索，了解它们是如何 他们会过度饮酒。 好了！

PROJECT SUMMARY Alcohol use disorder is a major public health problem. To date, available treatment options are limited to psychosocial intervention, three FDA approved medications (disulfuram, acamprosate, and naltrexone) and a few drugs approved for other indications, all with relatively small effect sizes. There is a clear need for additional treatments based on a deeper understanding of neurobiological mechanisms. Corticotrophin releasing factor (CRF) is a 41-amino acid neuropeptide produced mainly by neurons of the paraventricular hypothalamus, central amygdala (CeA), and bed nucleus of the stria terminalis (BNST) where it plays an important role in behavioral and physiological responses to stress. CRF has been long implicated in driving excessive ethanol consumption through prior studies that used CRF receptor antagonists in rodents. However, recent attempts to test CRF receptor antagonists as treatments for alcohol craving in humans have been disappointing. Part of this lack of success may be due to inadequate drug-like properties of some compounds and by a need for different human laboratory models that test drug effects on withdrawal and negative reinforcement in dependent subjects. Another reason may be due to the fact that CRF is released from neurons with other peptides and neurotransmitters that may act in synergy to drive excessive drinking. Understanding which of these co-released factors is important necessitates a different strategy that focuses on the CRF neurons themselves rather than on CRF receptors. The lack of genetic access to subpopulations of CRF neurons has made it difficult to study the biology of CRF neurons, the sources of CRF in different brain regions, and the circuitry underlying CRF-regulated behaviors. To fill this gap, we generated a BAC transgenic Wistar rat line in which Cre recombinase is expressed from the Crh gene promoter to enable genetic access to CRF neurons. In this project, we will use these rats to pursue the hypothesis that as animals develop ethanol dependence, CRF neurons in the CeA and BNST promote ethanol consumption through the coordinated release of GABA, CRF, and other neuropeptides. We will examine this hypothesis by selectively activating or inhibiting these neuronal populations and their projections using chemogenetic tools, and we will address the relative importance of the different transmitters and modulators released from these neurons using Cre-dependent RNA interference. Finally, we will investigate the role of repeated ethanol consumption on the transcriptome of these CRF neuronal populations to understand how ethanol changes their phenotype, which should provide us with important new clues as to how they drive excessive drinking. !