CELLULAR MODELS OF DEPENDENCE USING BRAIN SLICES

使用脑切片的依赖性细胞模型

• 批准号: 6563146
• 负责人: GEORGE Robert SIGGINS
• 金额: $ 25.15万
• 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: 2001-12-01 至 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6563146
• 关键词: GABA receptor; NMDA receptors; alcoholism /alcohol abuse; alcoholism antagonist; amygdala; behavioral /social science research tag; disease /disorder model; drug withdrawal; ethanol; laboratory mouse; laboratory rat; model design /development; neuropharmacology; nucleus accumbens; reinforcer; relapse /recurrence; tissue /cell culture

This project proposes to continue cellular studies of the role of synaptic transmitters and their receptors in alcohol intoxication and addiction. The rationale behind this approach is the considerable literature showing the synapse as the most sensitive site of ethanol action, and particularly those mediated by glutamate and GABA release. Our past studies lead us to hypothesize a role for presynaptic opiate and GABA/B receptors and postsynaptic NMDA and GABA/A receptors in ethanol intoxication and dependence. Our rationale is also based on behavioral findings with abused drugs, including alcohol, suggesting that the nucleus accumbens (NAcc) and amygdala are key areas in the reinforcing properties of these drugs, and that these properties may involve the same transmitter candidates, among others. We propose four sets of experiments: 1) To examine the effects of chronic ethanol treatment (via the ethanol vapor inhalation method) and withdrawal on membrane and synaptic properties in NAcc core and shell and amygdala neurons; 2) To examine synaptic and membrane properties in NAcc core and shell and amygdala neurons in an animal model of 'relapse' or protracted abstinence, whereby rats are treated chronically with ethanol vapor and then withdrawn for 7-14 days (at a time when 'craving' would be maximal); 3) To continue studies of the effect of the 'anti-alcoholism' drug acamprosate on NMDA and GABA/b receptors in NA/cc neurons, but now in the relapse model of specific aim #2; 4) To initiate normative in vitro studies of acute and chronic ethanol on membrane and synaptic properties of accumbens neurons in the mouse, for later studies of murine 'relapse' and genetic models. These studies will use hippocampal, NA/cc and amygdala brain slices and involve standard intracellular and "patch-slice" whole- cell clamp methods. We will use the infrared videomicroscopic method to distinguish morphologically different cell types for comparison to electrophysiological properties. We will examine evoked, pharmaco- logically-isolated monosynaptic events, responses to exogenous transmitters, paired-pulse inhibition and spontaneous synaptic events, to test the specificity and site of ethanol action. We believe these studies will provide important new information on possible sequelae of ethanol dependence at the cellular level, and--by virtue of comparisons of ethanol and acamprosate actions in control and protracted abstinence models--will also provide clues as to the cellular and ion channel correlates of alcohol seeking behavior.

该项目建议继续对突触作用进行细胞研究 酒精中毒和成瘾中的递质及其受体。 这种方法背后的基本原理是大量文献表明 突触是乙醇作用最敏感的位点，特别是 那些由谷氨酸和 GABA 释放介导的。我们过去的研究引导我们 假设突触前阿片受体和 GABA/B 受体的作用 突触后 NMDA 和 GABA/A 受体在乙醇中毒和 依赖性。我们的理由还基于虐待行为的发现 药物，包括酒精，表明伏隔核（NAcc）和 杏仁核是增强这些药物特性的关键区域，并且 这些属性可能涉及相同的发射机候选者，其中 其他的。我们提出四组实验：1）检查效果 慢性乙醇治疗（通过乙醇蒸气吸入法）和 NAcc 核和壳中膜和突触特性的撤回 杏仁核神经元； 2) 检查 NAcc 中的突触和膜特性 “复发”动物模型中的核、壳和杏仁核神经元 长期禁欲，即长期用乙醇治疗大鼠 蒸气，然后撤回 7-14 天（此时“渴望”会出现） 最大）； 3）继续研究“反酗酒”的效果 药物阿坎酸作用于 NA/cc 神经元的 NMDA 和 GABA/b 受体，但现在 具体目标#2的复发模型； 4) 启动体外规范 急性和慢性乙醇对膜和突触特性的研究 小鼠伏隔神经元，用于小鼠“复发”的后续研究 和遗传模型。这些研究将使用海马、NA/cc 和杏仁核 脑切片并涉及标准细胞内和“补丁切片”整体 细胞钳方法。我们将使用红外视频显微方法 区分形态上不同的细胞类型以进行比较 电生理特性。我们将检查诱发的、药物的 逻辑上孤立的单突触事件，对外源性的反应 递质、配对脉冲抑制和自发突触事件， 测试乙醇作用的特异性和位点。我们相信这些研究 将提供有关乙醇可能产生的后遗症的重要新信息 细胞水平的依赖性，以及--通过与乙醇的比较 和阿坎酸在控制和长期禁欲模型中的行动——将 还提供了细胞和离子通道相关性的线索 寻求酒精的行为。