Electrophysiology of alcohol in extended amygdela

这个项目是基于杏仁中央核(CEA)和蓝斑的行为学发现 (LC)是参与应激反应和滥用药物增强特性的关键大脑区域， 这些行为可能涉及几种递质(GABA、谷氨酸、去甲肾上腺素)和神经肽。 (CRF、阿片类药物和甘丙素)。这两个区域都与动机行为和焦虑状态有关，而我们 假设CEA和LC内的这些相同的神经化学系统参与了过度的 在依赖的动物身上可以看到酒精饮料。因此，我们提出了几组实验：1)为了 通过比较CEA的细胞和网络功能，评估CRF受体在过量饮酒中的作用 在对照组和过量饮酒小鼠(WID模型)小鼠的脑片中，相对于乙醇 细胞化学联合增强GABA能IPSCs或抑制谷氨酸能EPSPS CRF及其受体的定位。2)确定kappa阿片受体(kappa opiate Receptor，KRs)在 过量饮酒，为了与我们的Mu和Delta受体数据进行比较，通过检测CEA细胞功能 大脑KRs基因敲除(KO)的WID小鼠的脑切片。3)确定甘丙素及其受体的作用 通过检测WID小鼠和WID小鼠切片中的CEA和LC细胞来检测过量饮酒中的受体 脑胆汁和Gal2受体以及甘丙肽过度表达的KOS，以及神经化学和 CEA和LC神经元的分子生物学测量。4)确定对最大的WID的影响- 由Crabbe和Crabbe选择性培育的HDID小鼠的特异性AIMS 1-3的结果诱导的变化 芬兰人在黑暗中酗酒与他们的对照组相比，以及Shag与SLAG品系，被选为 预定的高和低酒精消耗量。电生理学研究将使用CEA和LC脑 包括标准的细胞内和全细胞钳制方法。我们将采取一系列措施来 评估乙醇和多肽作用于突触前和突触后的部位。RIA、实时聚合酶链式反应和 受体结合研究将用于甘丙素研究。这个项目应该会提供重要的新的 在细胞水平上关于酒精中毒可能的后遗症的信息，以及通过比较 酒精和多肽在控制、过度饮酒和基因敲除模型中的作用也将提供线索，如 与酒精依赖的突触、细胞和离子通道相关。

This project is based on behavioral findings that the central amygdala nucleus (CeA) and locus coeruleus (LC) are key brain areas involved in stress reactions and the reinforcing properties of abused drugs, and that these behaviors may involve several transmitters (GABA, glutamate, norepinephrine) and neuropeptides (CRF, opioids and galanin). Both regions are implicated in motivated behaviors and anxiety states, and we hypothesize that these same neurochemical systems within the CeA and LC are involved in the excessive ethanol drinking seen in dependent animals. Therefore, we propose several sets of experiments: 1) To assess the role of CRF receptors in excessive drinking, by comparing the CeA cellular and network function in brain slices from control and excessively drinking mice (WID model) mice, with respect to the ethanol augmentation of GABAergic IPSCs or inhibition of glutamatergic EPSPs, combined with cytochemical localization of CRF and CRF receptors. 2) To determine the role of kappa opiate receptors (KORs) in excessive drinking, for comparison to our mu and delta receptor data, by examining CeA cellular function in brain slices from WID mice with a knockout (KO) for brain KORs. 3) To determine the role of galanin and its receptors in excessive drinking, by examining CeA and LC cellular in slices from WID mice and those with KOs for brain Gall and Gal2 receptors and with galanin over-expression, and by neurochemical and molecular biological measures in CeA and LC neurons. 4) To determine the effects on the largest WID- induced changes from the results of Specific Aims 1-3, in the HDID mice selectively bred by the Crabbe and Finn groups for high drinking in the dark versus their controls, and for SHAG vs.SLAG lines, selected for scheduled high and low alcohol consumption.The electrophysiological studies will use CeA and LC brain slices and involve standard intracellular and whole-cell clamp methods. We will use a battery of measures to assess the pre- versus postsynaptic sites of action of ethanol and peptide effects. RIA, real-time PCR and receptor binding studies will be used in the galanin studies. This project should provide important new ¿ information on the possible sequelae of ethanol intoxication at the cellular level, and, by comparisons of ethanol and peptide actions in control, excessively drinking, and knockout models, will also provide clues as to the synaptic, cellular and ion channel correlates of ethanol dependence.