Excessive drinking and urocortin 1 neurocircuit

描述(由申请人提供)：本提案寻求成为INIA联合体“过度饮酒的神经生物学基础”的一部分，并重点研究神经肽Urocortin 1(Ucn1)。Ucn1是促肾上腺皮质激素释放因子受体CRF1和CRF2最有效的内源性配体。Ucn1在脑内的主要来源是非节前Edinger-Westphal核(NpEW)。外侧隔(LS)是npEW的主要投影区之一。最近的证据表明，Ucn1系统对酒精极其敏感，该系统的差异使动物容易受到酒精消费的差异，并且该系统的操作调节酒精摄入量。基于这一证据，我们假设Ucn1活性的差异是过量饮酒的重要决定因素。在这个项目中，我们建议通过合作努力来研究三个具体的目标：(1)利用微阵列技术寻找在选择性繁殖的高酒精和低酒精消耗动物中npEW和LS中持续差异表达的基因。将探索以下动物模型：为在黑暗中过量饮酒而选择性繁殖的小鼠，为在预定液体获取程序中过度饮酒而选择性繁殖的小鼠，使用2瓶选择程序选择性繁殖的小鼠，使用2瓶选择程序选择性繁殖的大鼠，以及它们各自的对照品系。基因表达的差异将通过免疫组织化学、原位杂交和定量RT-PCR来证实。(2)使用三种行为模型测试Ucn1基因敲除小鼠的饮酒量：DID-在黑暗中过量饮酒；Shag-由于预定时间进入而过度饮酒；以及标准的两瓶饮酒程序。我们还将使用微阵列技术来研究Ucn1基因敲除小鼠是否在特定目标1中识别的基因中获得补偿。(3)测试在特定目标1和特定目标2中过度饮酒的动物模型中识别的基因是否在人死后大脑的npEW和LS中表达，以及它们在酒精受试者和对照组之间是否存在差异表达。人类识别基因的同源物将通过定量RT-PCR进行测试。综上所述，这些研究将提供对Ucn1神经回路及其参与过度饮酒的全面分析，并可能为开发俄勒冈州健康治疗酒精中毒和酒精滥用障碍的新方法奠定基础。

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.