Cross K M, Dahm L M, Bowers C W
Division of Neurosciences, Beckman Research Institute of the City of Hope, Duarte, California 91010-3011, USA.
In Vitro Cell Dev Biol Anim. 2000 Jan;36(1):50-7. doi: 10.1290/1071-2690(2000)036<0050:SMOCAI>2.0.CO;2.
Simple methods are presented for quantitating contraction and intracellular calcium simultaneously in single, cultured smooth muscle cells. These methods are the first to demonstrate that reliable velocities of cell shortening can be measured in cultured smooth muscle cells and that cells in vitro exhibit shortening velocities comparable to those measured in the fastest phasic muscles in situ. Temporal relationships between changes in intracellular calcium and shortening within single cells were determined with a resolution of 100 ms and were consistent with measures in more "classical" preparations. Intracellular calcium rose quickly and transiently 10-fold above the basal level of 80-90 nM in response to the muscarinic agonist, carbachol. Shortening of the cells occurred 200 ms after intracellular calcium began to rise. The sensitivity and reliability of these methods allowed the effects of different stimuli to be easily resolved. The present report demonstrates that genuine contractility need not be ignored in cultured smooth muscle cells and that the temporal relations between shortening and intracellular calcium mobilization can be quantitatively assessed in controlled in vitro environments.
本文介绍了在单个培养的平滑肌细胞中同时定量收缩和细胞内钙的简单方法。这些方法首次证明,在培养的平滑肌细胞中可以测量到可靠的细胞缩短速度,并且体外培养的细胞表现出的缩短速度与在最快的相位性肌肉原位测量的速度相当。在单个细胞内,细胞内钙变化与缩短之间的时间关系以100毫秒的分辨率确定,并且与在更“经典”制剂中的测量结果一致。响应毒蕈碱激动剂卡巴胆碱,细胞内钙迅速且短暂地升高至高于80 - 90 nM基础水平的10倍。细胞缩短在细胞内钙开始升高后200毫秒出现。这些方法的灵敏度和可靠性使得不同刺激的效果能够轻松分辨。本报告表明,在培养的平滑肌细胞中,真正的收缩性不容忽视,并且在受控的体外环境中,可以定量评估缩短与细胞内钙动员之间的时间关系。