Electrophysiology of alcohol in extended amygdala

扩展杏仁核中酒精的电生理学

• 批准号: 7214012
• 负责人: GEORGE Robert SIGGINS
• 金额: $ 29.78万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-27 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7214012
• 关键词: GABA receptor; NMDA receptors; alcoholic beverage consumption; alcoholism /alcohol abuse; amygdala; behavioral /social science research tag; cooperative study; corticotropin releasing factor; craving; drug /alcohol abstinence; drug withdrawal; electrophysiology; ethanol; gene targeting; genetically modified animals; glutamate receptor; hormone receptor; laboratory mouse; membrane potentials; nucleus accumbens; opioid receptor; receptor expression; synapses; video microscopy; voltage /patch clamp

DESCRIPTION (provided by applicant): 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) T 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 WIDinduced 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 squeal 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.

描述（由申请人提供）：该项目基于行为发现，即中央杏仁核（CeA）和蓝斑（LC）是参与应激反应和滥用药物的强化特性的关键大脑区域，并且这些行为可能涉及多种递质（GABA、谷氨酸、去甲肾上腺素）和神经肽（CRF、阿片类药物和甘丙肽）。这两个区域都与动机行为和焦虑状态有关，我们假设 CeA 和 LC 内的这些相同的神经化学系统与依赖性动物的过度饮酒有关。因此，我们提出了几组实验：1）通过比较对照小鼠和过度饮酒小鼠（WID模型）小鼠脑切片中的CeA细胞和网络功能，结合GABA能IPSC的乙醇增强或谷氨酸能EPSP的抑制，结合CRF和CRF受体的细胞化学定位，评估CRF受体在过度饮酒中的作用。 2) 为了确定 kappa 阿片受体 (KOR) 在过度饮酒中的作用，与我们的 mu 和 delta 受体数据进行比较，通过检查脑 KOR 敲除 (KO) 的 WID 小鼠脑切片中的 CeA 细胞功能。 3) 为了确定甘丙肽及其受体在过度饮酒中的作用，通过检查 WID 小鼠和脑 Gall 和 Gal2 受体 KO 以及甘丙肽过度表达的小鼠切片中的 CeA 和 LC 细胞，并通过 CeA 和 LC 神经元的神经化学和分子生物学测量。 4) 为了确定特定目标 1-3 结果对最大 WID 诱导变化的影响，在由 Crabbe 和 Finn 组选择性培育的 HDID 小鼠中，用于在黑暗中大量饮酒与对照，以及针对预定的高和低饮酒量选择的 SHAG 与 SLAG 品系。电生理学研究将使用 CeA 和 LC 脑切片，并涉及标准的细胞内和全细胞钳方法。我们将使用一系列措施来评估乙醇和肽效应的突触前和突触后作用位点。 RIA、实时 PCR 和受体结合研究将用于甘丙肽研究。该项目应该提供关于细胞水平上可能出现的乙醇中毒尖叫的重要新信息，并且通过比较对照模型、过度饮酒模型和基因敲除模型中乙醇和肽的作用，还将提供关于乙醇依赖的突触、细胞和离子通道相关性的线索。