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Am J Physiol Heart Circ Physiol 279: H1185-H1193, 2000;
0363-6135/00 $5.00
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Vol. 279, Issue 3, H1185-H1193, September 2000

H2O2 mediates Ca2+- and MLC20 phosphorylation-independent contraction in intact and permeabilized vascular muscle

Nancy J. Pelaez1, Tracey R. Braun1, Richard J. Paul3, Richard A. Meiss2, and C. Subah Packer1

1 Departments of Physiology and Biophysics and 2 Obstetrics and Gynecology, Indiana University School of Medicine, Indianapolis, Indiana 46202-5102; and 3 Department of Molecular and Cell Physiology, University of Cincinnati College of Medicine, Cincinnati, Ohio 45267-0576

One purpose of the current study was to establish whether vasoconstriction occurs in all vessel types in response to H2O2. Isometric force was measured in pulmonary venous and arterial rings, and isobaric contractions were measured in mesenteric arteries and veins in response to H2O2. A second purpose was to determine whether H2O2-induced contraction is calcium independent. The addition of H2O2 to calcium-depleted (using the Ca2+ ionophore ionomycin in zero calcium EGTA buffer) muscle caused contraction. Furthermore, permeabilized muscle contracted in response to H2O2 even in zero Ca2+. The final purpose was to determine whether the 20-kDa regulatory myosin light chain (MLC20) phosphorylation plays a role in H2O2-induced contraction. Pulmonary arterial strips were freeze-clamped at various time points during H2O2-induced contractions, and the relative amounts of phosphorylated MLC20 were measured. H2O2 caused dose-dependent contractions that were independent of MLC20 phosphorylation. ML-9, a myosin light chain kinase inhibitor, had no effect on the H2O2 contractile response. In conclusion, H2O2 induces Ca2+- and MLC20 phosphorylation-independent contraction in pulmonary and systemic arterial and venous smooth muscle.

reactive oxygen species; vasoconstriction; signal transduction; arterial smooth muscle; venous smooth muscle; myosin light chain


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