MECHANISMS OF OPIATE/OPIOID PEPTIDE ACTION

这些研究的长期目标是通过 哪种阿片剂和阿片肽通过改变正常神经元功能 神经系统培养细胞中受体介导事件的分析 起源。我们建议使用神经肿瘤细胞和感觉神经元 拥有Mu、Kappa和Delta受体研究急性和慢性 阿片剂和阿片肽的作用，这样我们最终就可以设计出一种 对毒品成瘾的合理治疗。 阿片类药物(0-30分钟)对神经母细胞瘤的急性影响将被研究。 表达两者的中国仓鼠神经元杂交细胞系(NCB-20) (脑啡肽)和西格玛样(苯吗啡)阿片受体和 电压敏感钙通道(VSCC)和大鼠嗜铬细胞瘤 (PC12)表达VSCC的细胞，释放神经递质，如 去甲肾上腺素(NE)，可被NGF诱导表达 脑啡肽受体。将特别强调以下方面的作用 阿片类药物调节内钙动员中的钙离子流动 AMP依赖蛋白磷酸化，钙依赖蛋白 磷酸化、聚磷脂酰肌醇周转、三磷酸肌醇 K+通量的释放和调制。我们还将使用背根神经节 由细胞去极化引起的P物质释放的细胞 被阿片类激动剂阻断。 我们的主要假设是阿片类药物诱导的细胞数量减少 环磷酸腺苷水平可能导致蛋白的磷酸化水平降低。 这些细胞对二氢吡啶敏感的钙通道。这将导致 减少引起的钙离子内流和随后的抑制 发射器释放。

The long-range goal of these studies is to understand the mechanisms by which opiates and opioid peptides alter normal neuronal function by analyzing receptor-mediated events in cultured cells of nervous system origin. We propose to use neurotumor cells and sensory neurons that possess Mu, Kappa and Delta receptors to study both the acute and chronic effects of opiates and opioid peptides so that eventually we can devise a rational therapy for narcotic addiction. Acute effects of opiates (0-30min) will be studied in a neuroblastoma x Chinese hamster neuron hybrid cell line (NCB-20) which expresses both delta (enkephalin) and sigma-like (benzomorphan) opiate receptors and voltage-sensitive calcium channels (VSCC), and in rat pheochromocytoma (PC12) cells which express VSCC, release neurotransmitters such as norepinephrine (NE), and which can be induced with NGF to express enkephalin receptors. Particular emphasis will be placed on the role of opiates in regulating calcium fluxes internal calcium mobilization, cyclic AMP-dependent protein phosphorylation, calcium-dependent protein phosphorylation, polyphosphoinositide turnover, inositol triphosphate release and modulation of K+ flux. We will also use dorsal root ganglion cells in which substance P release, evoked by cell depolarization, is blocked by opiate agonists. It is our primary hypothesis that an opiate-induced reduction of cellular cyclic AMP levels may result in a reduced phosphorylation of a dihydropyridine-sensitive calcium channel in these cells. This would lead to a reduction in evoked influx of calcium ions and subsequent inhibition of transmitter release.