毒蕈碱受体与人类神经母细胞瘤细胞系SK-N-SH中的磷酸肌醇水解和胞质钙升高相关。
Muscarinic receptors coupled to phosphoinositide hydrolysis and elevated cytosolic calcium in a human neuroblastoma cell line SK-N-SH.
作者信息
Baird J G, Lambert D G, McBain J, Nahorski S R
机构信息
Department of Pharmacology and Therapeutics, Leicester.
出版信息
Br J Pharmacol. 1989 Dec;98(4):1328-34. doi: 10.1111/j.1476-5381.1989.tb12681.x.
Abstract
- The effects of the muscarinic agonist carbachol on phosphoinositide metabolism and its relationship to alteration of intracellular calcium were examined in SK-N-SH human neuroblastoma cells. Muscarinic receptors on these cells are coupled to phospholipase C and the myo [2-3H]-inositol phosphates resulting from receptor activation of cells labelled with [3H]-inositol accumulate rapidly. The breakdown of both inositol monophosphate (InsP1) and inositol bisphosphate (InsP2) is sensitive to lithium with inhibition of the latter only observed at higher concentrations of this ion. 2. Use of the calcium indicator dye Fura 2 revealed that carbachol stimulates a biphasic increase in intracellular calcium. 3. Carbachol was able to stimulate both [3H]-inositol phosphate production and intracellular calcium levels with respective EC50 values of 15.9 +/- 1.0 microM and 10.7 +/- 3.2 microM, indicating that no amplification occurs between these steps in the signal transduction pathway. 4. Inositol 1,4,5 trisphosphate (Ins(1,4,5)P3) released 45Ca2+ in a stereospecific and dose-related manner from intracellular stores of permeabilised cells. 5. These results suggest that this cell line may represent a useful model system to investigate receptor-mediated phosphoinositide metabolism and calcium homeostasis.
摘要
- 在SK-N-SH人神经母细胞瘤细胞中研究了毒蕈碱激动剂卡巴胆碱对磷酸肌醇代谢的影响及其与细胞内钙变化的关系。这些细胞上的毒蕈碱受体与磷脂酶C偶联，用[3H] - 肌醇标记的细胞经受体激活后产生的肌醇[2-3H] - 磷酸迅速积累。肌醇单磷酸（InsP1）和肌醇二磷酸（InsP2）的分解对锂敏感，仅在该离子的较高浓度下才观察到对后者的抑制作用。2. 使用钙指示剂染料Fura 2显示，卡巴胆碱刺激细胞内钙呈双相增加。3. 卡巴胆碱能够刺激[3H] - 肌醇磷酸的产生和细胞内钙水平，其各自的EC50值分别为15.9±1.0 microM和10.7±3.2 microM，表明在信号转导途径的这些步骤之间没有发生放大作用。4. 肌醇1,4,5 - 三磷酸（Ins(1,4,5)P3）以立体特异性和剂量相关的方式从通透细胞的细胞内储存中释放45Ca2+。5. 这些结果表明，该细胞系可能代表一个有用的模型系统，用于研究受体介导的磷酸肌醇代谢和钙稳态。
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