Intracellular calcium in canine cultured tracheal smooth muscle cells is regulated by M3 muscarinic receptors
Intracellular calcium in canine cultured tracheal smooth muscle cells is regulated by M3 muscarinic receptors
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犬培养的气管平滑肌细胞中的细胞内钙受 M3 毒蕈碱受体的调节
DOI：
10.1111/j.1476-5381.1993.tb13910.x
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发表时间：
1993
影响因子：
7.3
通讯作者：
Yen‐Yi Wang
中科院分区：
文献类型：
--
作者：
Chuen;Y. Yo;Yen‐Yi Wang
1在负载fura-2的犬培养的气管平滑肌细胞（TSMC）中直接测量了暴露于卡巴胆碱期间细胞溶质Ca 2+浓度（[Ca 2 +]i）的调节。通过卡巴胆碱刺激毒蕈碱胆碱受体（毒蕈碱AChR）产生[Ca 2 +]i的剂量依赖性升高，随后是稳定的平台期。卡巴胆碱的峰值和持续平台反应的EC 50值分别为0.34和0.33 μm。2阿托品（10 μm）阻止了卡巴胆碱的所有反应，当在卡巴胆碱反应期间加入时，在5 s内显著但不完全降低[Ca 2 +]i。因此，[Ca 2 +]i的变化由卡巴胆碱介导的毒蕈碱AChRs。3 AF-DX 116（一种选择性M2拮抗剂）和4-二苯基乙酰氧基-N-甲基哌啶（4-DAMP，一种选择性M3拮抗剂）抑制卡巴胆碱刺激的[Ca 2 +]i增加，pKB值分别为6.4和9.4，对应于AF-DX 116拮抗该反应的低亲和力和4-DAMP拮抗该反应的高亲和力。[Ca ~（2+）]i的平台升高依赖于外源性Ca ~（2+）的存在。通过加入2 mm EGTA去除Ca 2+导致[Ca 2 +]i迅速下降至静息水平。在没有外部Ca 2+的情况下，只看到[Ca 2 +]i的初始瞬时峰值，然后下降到静息水平;然后在卡巴胆碱的持续存在下，通过添加Ca 2+（1.8 mm）可以诱发[Ca 2 +]i的持续升高。5 [Ca 2 +]i的变化需要Ca 2+内流，因为Ca 2+通道阻滞剂地尔硫卓（10 μ m）、硝苯地平（10 μ m）、维拉帕米（10 μm）和Ni 2+（5 mm）可降低卡巴胆碱引起的[Ca 2 +]i的初始和持续升高。这些Ca 2+通道阻滞剂在平台期应用时也降低了[Ca 2 +]i的持续升高。6总之，我们已经证明，卡巴胆碱刺激的[Ca 2 +]i最初可检测到的增加是由于Ca 2+从内部储存中释放，然后是外部Ca 2+流入细胞。这种细胞外Ca 2+的流入部分涉及L-型Ca 2 +-通道。M3毒蕈碱受体似乎介导犬TSMCs中的Ca 2+动员。
1 The regulation of cytosolic Ca2+ concentrations ([Ca2+]i) during exposure to carbachol was measured directly in canine cultured tracheal smooth muscle cells (TSMCs) loaded with fura‐2. Stimulation of muscarinic cholinoceptors (muscarinic AChRs) by carbachol produced a dose‐dependent rise in [Ca2+]i which was followed by a stable plateau phase. The EC50 values of carbachol for the peak and sustained plateau responses were 0.34 and 0.33 μm, respectively. 2 Atropine (10 μm) prevented all the responses to carbachol, and when added during a response to carbachol, significantly, but not completely decreased [Ca2+]i within 5 s. Therefore, the changes in [Ca2+]i by carbachol were mediated through the muscarinic AChRs. 3 AF‐DX 116 (a selective M2 antagonist) and 4‐diphenylacetoxy‐N‐methylpiperidine (4‐DAMP, a selective M3 antagonist) inhibited the carbachol‐stimulated increase in [Ca2+]i with pKB values of 6.4 and 9.4, respectively, corresponding to low affinity for AF‐DX 116 and high affinity for 4‐DAMP in antagonizing this response. 4 The plateau elevation of [Ca2+]i was dependent on the presence of external Ca2+. Removal of Ca2+ by the addition of 2 mm EGTA caused the [Ca2+]i to decline rapidly to the resting level. In the absence of external Ca2+, only an initial transient peak of [Ca2+]i was seen which then declined to the resting level; the sustained elevation of [Ca2+]i could then be evoked by the addition of Ca2+ (1.8 mm) in the continued presence of carbachol. 5 Ca2+ influx was required for the changes of [Ca2+]i, since the Ca2+‐channel blockers, diltiazem (10 μm), nifedipine (10 μm), verapamil (10 μm) and Ni2+ (5 mm), decreased both the initial and sustained elevation of [Ca2+]i in response to carbachol. These Ca2+‐channel blockers also decreased the sustained elevation of [Ca2+]i when applied during the plateau phase. 6 In conclusion, we have demonstrated that the initial detectable increase in carbachol‐stimulated [Ca2+]i is due to the release of Ca2+ from internal stores, followed by the flux of external Ca2+ into the cells. This influx of extracellular Ca2+ partially involves an L‐type Ca2+‐channel. M3 muscarinic receptors appear to mediate the Ca2+ mobilization in canine TSMCs.
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DOI：
10.1113/jphysiol.1989.sp017602
发表时间：
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期刊：
The Journal of physiology
影响因子：
--
作者：
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DOI：
--
发表时间：
1991
期刊：
The Journal of pharmacology and experimental therapeutics
影响因子：
--
作者：
Shieh,CC;Petrini,MF;Dwyer,TM;Farley,JM
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Farley,JM
DOI：
10.1073/pnas.81.21.6899
发表时间：
1984-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
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作者：
BARON, CB;CUNNINGHAM, M;COBURN, RF
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COBURN, RF
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DOI：
10.1152/ajplung.1993.264.1.l53
发表时间：
1993
期刊：
The American journal of physiology
影响因子：
--
作者：
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通讯作者：
Gunst,SJ
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DOI：
10.1152/ajpcell.1987.253.6.c817
发表时间：
1987
期刊：
The American journal of physiology
影响因子：
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作者：
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通讯作者：
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