MECHANISMS OF OPIATE/OPIOID PEPTIDE ACTION

这些研究的长期目标是通过以下方式了解其机制： 阿片类药物和阿片肽改变正常的神经元功能， 分析神经系统培养细胞中受体介导的事件 起源 我们建议使用神经瘤细胞和感觉神经元， 拥有Mu，Kappa和Delta受体，可用于研究急性和慢性 阿片类药物和阿片肽的影响，这样我们最终可以设计一个 麻醉品成瘾的合理治疗 阿片类药物的急性作用（0- 30分钟）将在神经母细胞瘤中进行研究。 中国仓鼠神经元杂交细胞系（NCB-20），表达δ （脑啡肽）和σ样（苯并吗啡喃）阿片受体， 电压敏感性钙通道（VSCC）和大鼠嗜铬细胞瘤 表达VSCC的PC 12细胞释放神经递质， 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： 阿片类药物调节钙流动，内部钙动员，循环 AMP依赖蛋白磷酸化，钙依赖蛋白 磷酸化，多磷酸肌醇周转，三磷酸肌醇 释放和调节K+通量。 我们还将使用背根神经节 细胞中，由细胞去极化引起的P物质释放， 被阿片激动剂阻断 我们的主要假设是，阿片类药物诱导的细胞减少 环AMP水平可能会导致一个磷酸化的减少， 二氢吡啶敏感的钙通道。 这将导致 引起的钙离子内流减少和随后的抑制 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.