CELLULAR NEUROBIOLOGY RESEARCH PROJECT

细胞神经生物学研究项目

• 批准号: 6719833
• 负责人: GEORGE Robert SIGGINS
• 金额: $ 27.71万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-03-13 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6719833
• 关键词: GABA receptor; NMDA receptors; alcoholism /alcohol abuse; alcoholism antagonist; amygdala; behavioral /social science research tag; disease /disorder model; drug withdrawal; electrophysiology; ethanol; laboratory rat; model design /development; neurobiology; neuropeptide Y; neuropharmacology; nucleus accumbens; reinforcer; relapse /recurrence; synapses; tissue /cell culture; voltage /patch clamp

DESCRIPTION (provided by applicant): This project proposes to continue cellular studies of the role of neurotransmitters and their receptors in alcohol intoxication and addiction, with the rationale that the synapse is the most sensitive site of ethanol action, and particularly those synapses mediated by glutamate, GABA, and neuropeptide release. Our past studies led us to hypothesize a role for 'metabotropic' receptors and postsynaptic NMDA and GABAA receptors in ethanol intoxication and dependence. A second rationale is based on behavioral findings suggesting that the extended amygdala is a key system in the addictive properties of several drugs, including alcohol, and that several transmitters, neuropeptides, and endocannabinoids may be involved there. The amygdala has been implicated in motivated behaviors and anxiety states. Stressors and anxiety-provoking stimuli may trigger relapse in human alcoholics, and ethanol withdrawal is associated with increases both in anxiety-like behavior and in ethanol self-administration in rodents. Therefore, we hypothesize that the same neuropharmacological systems within the extended amygdala's circuitry mediate increases in anxiety state and in ethanol self-administration that occur during withdrawal from chronic ethanol, and we propose the following 3 sets of cellular studies to test this hypothesis: 1) Examination of the effects and interactions of acute and chronic ethanol administration, early withdrawal and protracted abstinence on CRF effects in central amygdala (CeA) neurons. 2) Examination of the effects of acute and chronic ethanol administration, early withdrawal and abstinence on neuropeptide Y (NPY) effects in CeA neurons. 3) Testing the effects of acute and chronic ethanol, early withdrawal and abstinence on endocannabinoid effects in CeA and nucleus accumbens (NAcc). In all 3 of these aims, subjects will be sham (control) male rats or those receiving chronic ethanol either via ethanol vapor inhalation and/or via self-administration, and their CeA or NAcc sliced and studied 1-12 hours or 1-2 weeks after withdrawal. We will record from amygdala and NAcc brain slices with intracellular (current- and voltageclamp) and "patch-slice" whole-cell clamp methods. The infrared DIC-videomicroscopic method will be used to identify morphologically distinct cells types for comparison of electrophysiological and pharmacological properties. We will record evoked, pharmacologically-isolated monosynaptic currents or potentials, and spontaneous and miniature synaptic events, to better test the specificity and site of action of ethanol and ligands. We believe these studies will provide important new information on possible sequelae of ethanol intoxication at the cellular level, and, by virtue of analyses of ethanol and peptide/cannabinoid interactions in control, chronic and protracted abstinence models, will also provide clues as to the cellular and ion channel correlates of ethanol dependence.

描述（由申请人提供）：该项目建议继续对神经递质及其受体在酒精中毒和成瘾中的作用进行细胞研究，其基本原理是突触是乙醇作用最敏感的部位，特别是那些由谷氨酸介导的突触、GABA 和神经肽释放。我们过去的研究使我们推测“代谢型”受体以及突触后 NMDA 和 GABAA 受体在乙醇中毒和依赖中的作用。第二个基本原理是基于行为研究结果，表明扩展的杏仁核是包括酒精在内的多种药物成瘾特性的关键系统，并且可能涉及多种递质、神经肽和内源性大麻素。杏仁核与动机行为和焦虑状态有关。压力源和引起焦虑的刺激可能会引发人类酗酒者的复发，而乙醇戒断与啮齿类动物焦虑样行为和乙醇自我给药的增加有关。因此，我们假设扩展杏仁核回路内的相同神经药理学系统介导了长期乙醇戒断期间发生的焦虑状态和乙醇自我给药的增加，并且 我们提出以下三组细胞研究来检验这一假设：1）检查急性和慢性乙醇给药、早期戒断和长期戒酒对中央杏仁核（CeA）神经元CRF影响的影响和相互作用。 2)检查急性和慢性乙醇给药、早期戒断和戒酒对CeA神经元中神经肽Y(NPY)作用的影响。 3) 测试急性和慢性乙醇、早期戒断和戒断对 CeA 和伏隔核 (NAcc) 内源性大麻素影响的影响。在所有这 3 个目标中，受试者将是假（对照）雄性大鼠或通过乙醇蒸气吸入和/或通过自我给药接受慢性乙醇的大鼠，并将其 CeA 或 NAcc 切片并研究 1-12 小时或 1-2撤药后几周。我们将使用细胞内（电流和电压钳）记录杏仁核和 NAcc 脑切片 和“贴片切片”全细胞钳方法。红外 DIC-视频显微方法将 用于识别形态上不同的细胞类型，以比较电生理学和 药理特性。我们将记录诱发的、药理学隔离的单突触电流或电位，以及自发和微型突触事件，以更好地测试乙醇和配体的特异性和作用位点。我们相信这些研究将为细胞水平上乙醇中毒可能的后遗症提供重要的新信息，并且通过在对照、慢性和长期戒断模型中对乙醇和肽/大麻素相互作用的分析，还将提供关于细胞水平的线索。和离子通道与乙醇依赖性相关。