Excessive drinking and urocortin 1 neurocircuit

描述（由申请人提供）：该提案旨在成为INIA联盟“过量饮酒的神经生物学基础”的一部分，并关注神经肽Urocortin 1（Ucn 1）。Ucn 1是促肾上腺皮质激素释放因子（CRF）受体CRF 1和CRF 2的最有效的内源性配体。Ucn 1在脑中的主要来源是非节前Edinger-Westphal核（npEW）。npEW的主要投射区域之一是外侧隔（LS）。最近的证据表明，Ucn 1系统是非常敏感的酒精，在这个系统的差异易使动物的差异，酒精消费，并操纵这个系统调节酒精摄入量。基于这一证据，我们假设Ucn 1活性的差异是过量饮酒的重要决定因素。在本项目中，我们提出了三个具体的目标：（1）使用微阵列技术来识别选择性饲养的高和低酒精消耗动物之间在npEW和LS中表现出一致差异表达的基因。将探索以下动物模型：在黑暗中选择性饲养过量饮酒的小鼠、在预定液体进入程序中选择性饲养过量饮酒的小鼠、使用2瓶选择程序选择性饲养的小鼠、使用2瓶选择程序选择性饲养的大鼠及其各自的对照系。将使用免疫组织化学、原位杂交和定量RTPCR确认基因表达差异。(2)使用三种行为模型测试Ucn 1基因敲除小鼠的酒精消耗：DID -在黑暗中过度饮酒; SHAG -由于预定访问而过度饮酒;以及标准的2瓶饮酒程序。我们还将使用微阵列技术来研究Ucn 1基因敲除小鼠是否在特定目标1中鉴定的基因中产生补偿。(3)为了测试在特定目标1和特定目标2中过量饮酒的动物模型中鉴定的基因是否在人类死后大脑的npEW和LS中表达，以及它们是否在酒精受试者和对照之间差异表达。将通过定量RT-PCR检测已鉴定基因的人类同源物。综上所述，这些研究将提供一个彻底的全面分析Ucn 1神经回路及其参与过量饮酒，并可能提供基础的新方法的发展俄勒冈州健康治疗酒精中毒和酒精滥用障碍。

DESCRIPTION (provided by applicant): This proposal seeks to become a part of the INIA consortium "Neurobiologial Basis of Excessive Drinking", and focuses of the neuropeptide Urocortin 1 (Ucn1). Ucn1 is the most effective endogenous ligand of both corticotropin releasing factor (CRF) receptors CRF1 and CRF2. The main source of Ucn1 in the brain is the non-preganglionic Edinger-Westphal nucleus (npEW). One of the main projection areas of npEW is the lateral septum (LS). Recent evidence indicates that the Ucn1 system is extremely sensitive to alcohol, that differences in this system predispose animals to differences in alcohol consumption, and that manipulations of this system regulate alcohol intake. Based on this evidence we hypothesize that differences in Ucn1 activity are important determinants of excessive alcohol intake. In this project we propose to apply collaborative efforts to investigate three specific aims: (1) To identify genes showing consistently different expression in npEWand LS between selectively-bred high and low alcohol consuming animals using microarray technology. Following animal models will be explored: mice selectively bred for excessive drinking in the dark, mice selectively bred for excessive drinking in the scheduled fluid access procedure, mice selectively bred using the 2-bottle choice procedure, rats selectively bred using the 2-bottle choice procedure, and their respective control lines. Differences in gene expression will be confirmed using immunohistochemistry, in situ hybridization and quantitative RTPCR. (2) To test alcohol consumption in Ucn1 knockout mice using three behavioral models: DID - excessive drinking in the dark; SHAG - excessive drinking due to scheduled access; and the standard 2-bottle drinking procedure. We will also use microarray technology to investigate whether Ucn1 knockout mice developed compensations in genes identified in Specific Aim 1. (3) To test whether genes identified in animal models of excessive alcohol consumption in Specific Aim 1 and Specific Aim 2 are expressed in npEW and LS of human post-mortem brains, and whether they are differentially expressed between alcoholic subjects and controls. Human homologues of the identified genes will be tested by quantitative RT-PCR. Taken together, these studies will provide a thorough comprehensive analysis of the Ucn1 neurocircuit and its involvement in excessive alcohol consumption, and could provide groundwork for development of new approaches for Oregon Health treatments of alcoholism and alcohol abuse disorders.