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Am J Physiol Cell Physiol 273: C1714-C1720, 1997;
0363-6143/97 $5.00
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Vol. 273, Issue 5, C1714-C1720, November 1997

Long-term regulation of contractility and calcium current in smooth muscle

Maria Gomez and Karl Swärd

Department of Physiology and Neuroscience, Lund University, S-223 62 Lund, Sweden

Longitudinal smooth muscle strips from guinea pig ileum were cultured in vitro for 5 days, and the relationship between extracellular Ca2+ and force in high-K+ medium was evaluated. In strips cultured with 10% fetal calf serum (FCS), this relationship was shifted to the right (50% effective concentration changed by 2-3 mM) compared with strips cultured without FCS. The shift was prevented by inclusion of verapamil (1 µM) during culture and mimicked by ionomycin in the absence of FCS. The intracellular Ca2+ concentration ([Ca2+]i) during stimulation with high-K+ solution or carbachol was reduced after culture with FCS, whereas the [Ca2+]i-force relationship was unaffected. Cells were isolated from cultured strips, and whole cell voltage-clamp experiments were performed. Maximum inward Ca2+ current (10 mM Ba2+), normalized to cell capacitance, was almost three times smaller in cells isolated from strips cultured with FCS. Culture with 1 µM verapamil prevented this reduction. These results suggest that increased [Ca2+]i during culture downregulates Ca2+ current density, with associated effects on contractility.

tissue culture; fura 2; patch clamp; verapamil; ionomycin


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