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Na+/K+/Cl− cotransport in cultured vascular smooth muscle cells: Stimulation by angiotensin II and calcium ionophores, inhibition by cyclic AMP and calmodulin antagonists

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Summary

The specific activity of the Na+/K+/Cl− cotransporter was assayed by measuring the initial rates of furosemide-inhibitable86Rb+ influx and efflux. The presence of all three ions in the external medium was essential for cotransport activity. In cultured smooth muscle cells furosemide and bumetanide inhibited influx by 50% at 5 and 0.2 μm, respectively. The dependence of furosemide-inhibitable86Rb+ influx on external Na+ and K+ was hyperbolic with apparentK m values of 46 and 4mm, respectively. The dependence on Cl− was sigmoidal. Assuming a stoichiometry of 1∶1∶2 for Na+/K+/Cl−, aK m of 78mm was obtained for Cl−. In quiescent smooth muscle cells cotransport activity was approximately equal to Na+ pump activity with each pathway accounting for 30% of total86Rb+ influx. Growing muscle cells had approximately 3 times higher cotransport activity than quiescent ones. Na+ pump activity was not significantly different in the gorwing and quiescent cultures. Angiotensin II (ANG) stimulated cotransport activity as did two calcium-transporting ionophores, A23187 and ionomycin. The removal of external Ca2+ prevented A23187, but not ANG, from stimulating the cotransporter. Calmodulin antagonists selectively inhibited86Rb+ influx via the cotransporter. Beta-adrenoreceptor stimulation with isoproterenol, like other treatments which increase cAMP, inhibited cotransport activity. Cultured porcine endothelial cells had 3 times higher cotransport activity than growing muscle cells. Calmodulin antagonists inhibited cotransport activity, but agents which increase cAMP or calcium had no effect on cotransport activity in the endothelial cells.

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  1. Department of Pharmacology, University of Alabama at Birmingham, 35294, Birmingham, Alabama

    Jeffrey Bingham Smith & Lucinda Smith

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  1. Jeffrey Bingham Smith
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Smith, J.B., Smith, L. Na+/K+/Cl− cotransport in cultured vascular smooth muscle cells: Stimulation by angiotensin II and calcium ionophores, inhibition by cyclic AMP and calmodulin antagonists. J. Membrain Biol. 99, 51–63 (1987). https://doi.org/10.1007/BF01870621

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  • DOI: https://doi.org/10.1007/BF01870621

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