CELLULAR MODELS OF DEPENDENCE USING BRAIN SLICES

该项目建议继续对突触的作用进行细胞研究。 酒精中毒和成瘾中的递质及其受体。 这种方法背后的理论基础是大量的文献表明 突触是乙醇作用最敏感的部位，尤其是 谷氨酸和γ-氨基丁酸的释放。我们过去的研究引导我们 假设突触前阿片和GABA/B受体的作用 突触后NMDA和GABA/A受体与乙醇中毒 依赖。我们的理由也是基于对被虐待的行为的发现 药物，包括酒精，表明伏隔核(NAcc)和 杏仁核是这些药物增强性能的关键区域，并且 这些属性可能涉及相同的候选发送器，其中 其他。我们提出了四组实验：1)检验 慢性酒精治疗(通过酒精蒸气吸入法)和 NAcc核和壳中的膜和突触特性的戒断 杏仁核神经元；2)检测NAcc的突触和膜特性 复发性痴呆动物模型中的核壳和杏仁核神经元 长期禁欲，即用乙醇长期治疗大鼠 蒸气，然后退出7-14天(在一段时间，‘渴望’将 最大)；3)继续研究“解酒”的效果 药物氨基己酸酯对NA/CC神经元NMDA和GABA/b受体的影响 2；4)体外启动标准化的特定目标复发模型 急、慢性乙醇对细胞膜和突触特性的影响 对小鼠伏隔核神经元的研究，为以后研究小鼠的“复发”做准备 和遗传模型。这些研究将使用海马体、NA/CC和杏仁核。 脑片，并涉及标准的细胞内和“贴片切片”整体- 细胞钳制方法。我们将使用红外摄像显微镜的方法 区分形态上不同的细胞类型，以便与 电生理特性。我们会检查诱发的，药剂- 逻辑隔离的单突触事件，对外源的反应 成对脉冲抑制和自发突触事件的递质，以 检测乙醇作用的特异性和部位。我们相信这些研究 将为乙醇可能的后遗症提供重要的新信息 在细胞水平上的依赖性，以及--通过与乙醇的比较 在控制和长期戒断模型中采取积极的行动--Will 还提供了与细胞和离子通道相关的线索 寻酒行为。

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an animal model of 'relapse' or protracted abstinence, whereby rats are treated chronically with ethanol vapor and then withdrawn for 7-14 days (at a time when 'craving' would be maximal); 3) To continue studies of the effect of the 'anti-alcoholism' drug acamprosate on NMDA and GABA/b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.