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， 递质及其受体在酒精中毒和成瘾中的作用。 这种方法背后的基本原理是大量文献表明， 突触是乙醇作用的最敏感部位， 由谷氨酸和GABA释放介导。我们过去的研究使我们 假设突触前阿片和GABA/B受体作用， 突触后NMDA和GABA/A受体在乙醇中毒和 依赖我们的理论基础也是基于对虐待行为的研究结果， 包括酒精在内的药物，这表明，神经核（NAcc）和 杏仁核是这些药物增强特性的关键区域， 这些属性可能涉及相同的发射机候选者， 他人我们提出了四组实验：1）检查的影响， 长期乙醇治疗（通过乙醇蒸汽吸入法）， 在NAcc核和壳中的膜和突触特性上的撤回， 杏仁核神经元的突触和膜特性 核心和壳和杏仁核神经元在“复发”或 长期禁欲，即大鼠长期接受乙醇治疗 蒸汽，然后撤回7-14天（在“渴望”的时候， 最大）; 3）继续研究“戒酒”的效果 药物阿坎酸对NA/cc神经元中的NMDA和GABA/B受体的影响，但现在， 特定目标复发模型#2; 4）启动体外规范性研究 急性和慢性乙醇对细胞膜和突触特性的影响 小鼠的神经元，用于以后对小鼠“复发”的研究 基因模型。这些研究将使用海马、NA/cc和杏仁核 脑切片，并涉及标准的细胞内和“补丁切片”的整体， 细胞钳方法。我们将使用红外视频显微镜的方法， 区分形态上不同的细胞类型， 电生理特性我们会检查诱发的，药理学的- 逻辑上孤立的单突触事件，对外源性 传递，成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 将提供重要的新信息，对乙醇可能的后遗症， 在细胞水平上的依赖性，以及--通过比较乙醇 和阿坎酸在控制和长期禁欲模型中的作用-- 还提供了细胞和离子通道相关的线索， 酒精寻求行为

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animal model of 'relapse' or protracted abstinence, whereby rats are treated chronically with ethanol vapor and then withdrawn for 7-14 days (at a time when 'craving' would be maximal); 3) To continue studies of the effect of the 'anti-alcoholism' drug acamprosate on NMDA and GABA/b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