Published Online
on June 9, 2003

A more recent version of this article appeared on June 24, 2003

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Submitted on January 13, 2003
Revised on February 27, 2003
Accepted on March 12, 2003

Nitric Oxide-Induced Decrease in Calcium Sensitivity of Resistance Arteries Is Attributable to Activation of the Myosin Light Chain Phosphatase and Antagonized by the RhoA/Rho Kinase Pathway

Steffen-Sebastian Bolz MD^*, Lukas Vogel , Daniel Sollinger , Roland Derwand MD, Cor de Wit MD, Gervaise Loirand PhD, and Ulrich Pohl MD, PhD

From Physiologisches Institut, Ludwig Maximilians Universität, München, Germany, and INSERM U-533 (G.L.), Faculté des Sciences et Techniques, Nantes, France.

^* To whom correspondence should be addressed. E-mail: bolz{at}lrz.uni-muenchen.de.

Background--NO-induced dilations in resistance arteries (RAs) are not associated with decreases in vascular smooth muscle cell Ca²⁺. We tested whether a cGMP-dependent activation of the smooth muscle myosin light chain phosphatase (MLCP) resulting in a Ca²⁺ desensitization of the contractile apparatus was the underlying mechanism and whether it could be antagonized by the RhoA pathway.

Methods and Results--The Ca²⁺ sensitivity of RA was assessed as the relation between changes in diameter and [Ca²⁺]_i in depolarized RA (120 mol/L K⁺) exposed to stepwise increases in Ca²⁺_ex (0 to 3 mmol/L). Effects of 10 µmol/L sodium nitroprusside (SNP) on Ca²⁺ sensitivity were determined before and after application of the soluble guanylate cyclase inhibitor ODQ (1 µmol/L) and the MLCP inhibitor calyculin A (120 nmol/L) and in presence of the RhoA-activating phospholipid sphingosine-1-phosphate (S1P, 12 nmol/L). SNP-induced dilations were also studied in controls and in RAs pretreated with the Rho kinase inhibitor Y27632 or transfected with a dominant-negative RhoA mutant (N19RhoA). Constrictions elicited by increasing Ca²⁺_ex were significantly attenuated by SNP, which, however, left associated increases in [Ca²⁺]_i unaffected. This NO-induced attenuation was blocked by ODQ, calyculin A, and S1P. The S1P-induced translocation of RhoA indicating activation of the GTPase was not reversed by SNP. Inhibition of RhoA/Rho kinase by N19RhoA or Y27632 significantly augmented SNP-induced dilations.

Conclusions--NO dilates RA by activating the MLCP in a cGMP-dependent manner, thereby reducing the apparent Ca²⁺ sensitivity of the contractile apparatus. MLCP inactivation via the RhoA/Rho kinase pathway antagonizes this Ca²⁺-desensitizing effect that, in turn, can be restored using RhoA/Rho kinase inhibitors.

Key words: microcirculation • muscle, smooth • nitric oxide • signal transduction • vasodilation

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