Ca2+ oscillation and c-fos gene expression induced via muscarinic acetylcholine receptor in human T- and B-cell lines
Ca2+ oscillation and c-fos gene expression induced via muscarinic acetylcholine receptor in human T- and B-cell lines
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DOI：
10.1007/s002100000251
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发表时间：
2000-07-01
影响因子：
3.6
通讯作者：
Kawashima, K
中科院分区：
文献类型：
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作者：
Fujii, T;Kawashima, K
我们以前报道，血液中的乙酰胆碱（ACh）主要来源于T淋巴细胞和毒蕈碱（Ms）ACh受体mRNA的表达在T和B淋巴细胞。在本研究中，我们使用共聚焦激光扫描显微镜和fluo-3，一种钙敏感的指示剂，研究Ms-ACh受体激动剂对细胞内游离Ca 2+浓度（[Ca 2 +]（i））的影响，从人T细胞系（CEM）和B细胞系（Daudi）的单个细胞中，我们用作淋巴细胞模型。在两种细胞系中，用ACh刺激Ms-ACh受体，（0.1-100 μ M），氨甲酰甲胆碱（100 μ M）卡巴胆碱（100 μ M）或氧化震颤素-M（氧代-M：0.1-100 μ M）诱导Fluo-3荧光的[Ca 2 +]（i）依赖性增加，在存在细胞外Ca 2+的情况下，随后是[Ca 2 +]（i）的振荡，所有的作用都能被Ms-ACh受体拮抗剂阿托品（1 μ M）完全阻断。在这两个细胞系Oxo-M（100 μ M）上调c-fos mRNA的表达在细胞外Ca 2+依赖的方式。同样，该作用被1 μ M阿托品阻断。这些结果提供了第一个证据表明，刺激Ms-ACh受体诱导钙振荡和上调c-fos基因表达的T-和B-淋巴细胞，这是一致的概念，ACh从T-淋巴细胞释放触发核信号通过Ms-ACh受体。
We previously reported that blood acetylcholine (ACh) mainly originates from T-lymphocytes and that muscarinic (Ms) ACh receptor mRNA is expressed in both T- and B-lymphocytes. In the present study, we used confocal laser scanning microscopy and fluo-3, a calcium-sensitive indicator, to investigate the effects of Ms-ACh receptor agonists on the intracellular free Ca2+ concentration ([Ca2+](i)) in single cells from human T-cell (CEM) and B-cell (Daudi) lines, which we used as models of lymphocytes. In both cell lines, stimulation of Ms-ACh receptors with ACh (0.1-100 mu M), bethanechol (100 mu M) carbachol (100 mu M) or oxotremorine-M (Oxo-M: 0.1-100 mu M) induced [Ca2+](i)-dependent increases in fluo-3 fluorescence, which in the presence of extracellular Ca2+ were followed by oscillations in [Ca2+](i), that persisted For at least 10 min. All effects were completely blocked by atropine (1 mu M), an Ms-ACh receptor antagonist. In both cell lines Oxo-M (100 mu M) up-regulated expression of c-fos mRNA in an extracellular Ca2+-dependent manner. Again, the effect was blocked by 1 mu M atropine. These results provide the first evidence that stimulation of Ms-ACh receptors induces Ca2+ oscillations and up-regulates c-fos gene expression in T- and B-lymphocytes, which is consistent with the notion that ACh released from T-lymphocytes triggers nuclear signaling via Ms-ACh receptors.