EFFECTS OF MORPHINE AND ANALOGUES ON CELL METABOLISM

吗啡及其类似物对细胞代谢的影响

• 批准号: 3206697
• 负责人: Eric J Simon
• 金额: $ 47.69万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1975
• 资助国家: 美国
• 起止时间: 1975-12-01 至 1988-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3206697
• 关键词: Anura; affinity chromatography; affinity labeling; analgesics; autoradiography; bioassay; brain metabolism; cell membrane; chemical binding; conformation; drug metabolism; drug tolerance; electron spin resonance spectroscopy; enkephalins; enzyme linked immunosorbent assay; ethanol; fluorescent dye /probe; glycoproteins; guinea pigs; human tissue; immunofluorescence technique; laboratory mouse; laboratory rat; liposomes; membrane activity; membrane structure; microinjections; monoclonal antibody; morphine; naltrexone; narcotics; neuropeptides; opioid receptor; phospholipase A2; radioimmunoassay; spinal cord; tissue /cell culture

The long term objective of our research is the elucidation of the molecular mechanism of the acute and chronic effects of opiate narcotic analgesics. Our approach to this problem rests primarily on the study of the endogenous opioid peptide-opiate receptor system. A knowledge of the workings of this system would have wide-ranging implications and, we feel, will lead towards an understanding of the actions of exogenous opiates. Our focus will be on opiate receptors, their properties and isolation. The proposed research can be divided into 4 categories: 1) membrane-bound opiate receptors 2) soluble opiate receptors 3) opiate receptors and endogenous opioids in tissue culture and 4) involvement of the endogenous opioid system in the control of pain and aversion. In studies with membrane-bound receptors we will attempt further characterization of agonist and antagonist binding and of receptor subtypes by thermodynamic and kinetic experiments and selective inhibition or inactivation of receptor subclasses. In collaboration with its developers, Drs. Hirth and Goeldner, a novel method for covalent labeling called "energy transfer photoaffinity labeling" will be applied to opiate receptors. Good yields of solubilized opiate receptors have been obtained by us from nonmammalian and mammalian brain. We plan to characterize these by physical chemical methods and by reconstitution into liposomes and cell membranes. Purification of solubilized receptors and separation of receptor subtypes will be carried out. Attempts will be made to raise monoclonal antibodies, which would aid in further purification and make feasible molecular biological experiments. Tissue culture of spinal cord with attached dorsal root ganglia from fetal mice will be used for studies of opiate receptor subtypes and opioid peptides in tolerance and under various environmental conditions. Fluorescent enkephalin derivatives will be used in video-intensification fluorescence microscopic experiments to learn about receptor distribution and internalization. An approach begun about 2 years ago in this laboratory is the study of the role of endogenous opioids and opiate receptors in the regulation of pain and aversion, using behavioral and brain stimulation procedures. The role of opiate receptor subclasses and endogenous opioids in the modulation of pain responses organized at different levels of the CNS will be assessed.

我们研究的长期目标是阐明分子机制 阿片类麻醉镇痛药的急性和慢性作用的机制。 我们解决这个问题的方法主要依赖于内源性的研究 阿片肽-阿片受体系统。 了解此功能的工作原理 系统将产生广泛的影响，我们认为，将导致 了解外源性阿片类药物的作用。 我们的重点将是 阿片受体、其特性和分离。 拟议的研究 可分为4类：1）膜结合阿片受体2） 可溶性阿片受体 3) 阿片受体和内源性阿片类药物 组织培养和4）内源性阿片系统的参与 控制疼痛和厌恶。 在膜结合受体的研究中，我们 将尝试进一步表征激动剂和拮抗剂的结合 通过热力学和动力学实验以及选择性的受体亚型 受体亚类的抑制或失活。 与合作 它的开发者，博士。 Hirth 和 Goeldner，一种新的共价方法 称为“能量转移光亲和标记”的标记将应用于 阿片受体。 溶解阿片受体的良好收率 我们从非哺乳动物和哺乳动物的大脑中获得。 我们计划 通过物理化学方法和重构来表征它们 脂质体和细胞膜。 溶解的受体的纯化和 将进行受体亚型的分离。 将会做出尝试 产生单克隆抗体，这将有助于进一步纯化和 进行可行的分子生物学实验。 脊柱组织培养 带有来自胎儿小鼠的背根神经节的脐带将用于 阿片受体亚型和阿片肽的耐受性和耐受性研究 在各种环境条件下。 荧光脑啡肽衍生物 将用于视频增强荧光显微实验 了解受体分布和内化。 一种方法 大约两年前，这个实验室开始研究 内源性阿片类药物和阿片受体在疼痛调节中的作用 厌恶，使用行为和大脑刺激程序。 的作用 阿片受体亚类和内源性阿片类药物在疼痛调节中的作用 将评估中枢神经系统不同级别组织的应对措施。