Simultaneous measures of contraction and intracellular calcium in single, cultured smooth muscle cells |
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Authors: | Kathryn M L Cross Lisa M Dahm Chauncey W Bowers |
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Institution: | (1) Division of Neurosciences, Beckman Research Institute of the City of Hope, 1450 East Duarte Road, 91010-3011 Duarte, California |
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Abstract: | Summary 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. |
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Keywords: | amnion calcium transient defined media Indo-1 |
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