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Am J Physiol Lung Cell Mol Physiol 276: L35-L40, 1999;
1040-0605/99 $5.00
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Vol. 276, Issue 1, L35-L40, January 1999

cGMP modulation of Ca2+ sensitivity in airway smooth muscle

Keith A. Jones, Gilbert Y. Wong, Christopher J. Jankowski, Masaki Akao, and David O. Warner

Departments of Anesthesiology and Physiology and Biophysics, Mayo Clinic and Mayo Foundation, Rochester, Minnesota 55905

A beta -escin-permeabilized canine tracheal smooth muscle preparation was used to test the hypothesis that cGMP decreases Ca2+ sensitivity in airway smooth muscle primarily by inhibiting the membrane receptor-coupled mechanisms that regulate Ca2+ sensitivity and not by inhibiting Ca2+/calmodulin activation of the contractile proteins. 8-Bromo-cGMP (100 µM) had no effect on the free Ca2+ concentration-response curves generated in the absence of muscarinic receptor stimulation. In the presence of 100 µM ACh plus 10 µM GTP, 8-bromo-cGMP (100 µM) caused a rightward shift of the free Ca2+ concentration-response curve, significantly increasing the EC50 for free Ca2+ from 0.35 ± 0.03 to 0.75 ± 0.06 µM; this effect of 8-bromo-cGMP was concentration dependent from 1 to 100 µM. 8-Bromo-cGMP (100 µM) decreased the level of regulatory myosin light chain (rMLC) phosphorylation for a given cytosolic Ca2+ concentration but had no effect on the amount of isometric force produced for a given level of rMLC phosphorylation. These findings suggest that cGMP decreases Ca2+ sensitivity in canine tracheal smooth muscle primarily by inhibiting the membrane receptor-coupled mechanisms that modulate the relationship between cytosolic Ca2+ concentration and rMLC phosphorylation.

guanosine 3',5'-cyclic monophosphate; trachea; calcium sensitivity; myosin light chain phosphorylation; acetylcholine; guanosine 5'-O-(3-thiotriphosphate); fura 2; beta -escin; nitrovasodilators


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