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Originally published In Press as doi:10.1074/jbc.M211075200 on November 5, 2002

J. Biol. Chem., Vol. 278, Issue 3, 1856-1863, January 17, 2003
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Ca2+-independent Phospholipase A2 Is Required for Agonist-induced Ca2+ Sensitization of Contraction in Vascular Smooth Muscle*

Zhenheng GuoDagger , Wen SuDagger , Zhongmin Ma§, George M. SmithDagger , and Ming C. GongDagger

From the Dagger  Department of Physiology, University of Kentucky Medical Center, Lexington, Kentucky 40536 and the § Division of Experimental Diabetes and Aging, Department of Geriatrics and Adult Development, Mount Sinai School of Medicine, New York, New York 1002

Excitatory agonists can induce significant smooth muscle contraction under constant free Ca2+ through a mechanism called Ca2+ sensitization. Considerable evidence suggests that free arachidonic acid plays an important role in mediating agonist-induced Ca2+-sensitization; however, the molecular mechanisms responsible for maintaining and regulating free arachidonic acid level are not completely understood. In the current study, we demonstrated that Ca2+-independent phospholipase A2 (iPLA2) is expressed in vascular smooth muscle tissues. Inhibition of the endogenous iPLA2 activity by bromoenol lactone (BEL) decreases basal free arachidonic acid levels and reduces the final free arachidonic acid level after phenylephrine stimulation, without significant effect on the net increase in free arachidonic acid stimulated by phenylephrine. Importantly, BEL treatment diminishes agonist-induced Ca2+ sensitization of contraction from 49 ± 3.6 to 12 ± 1.0% (p < 0.01). In contrast, BEL does not affect agonist-induced diacylglycerol production or contraction induced by Ca2+, phorbol 12,13-dibutyrate (a protein kinase C activator), or exogenous arachidonic acid. Further, we demonstrate that adenovirus-mediated overexpression of exogenous iPLA2 in mouse portal vein tissue significantly potentiates serotonin-induced contraction. Our data provide the first evidence that iPLA2 is required for maintaining basal free arachidonic acid levels and thus is essential for agonist-induced Ca2+-sensitization of contraction in vascular smooth muscle.

* This work was supported by National Institutes of Health (NIH) Grant HL67284 (to M. C. G.) and by NINDS, NIH, Grant NS38126 (to G. M. S.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed. Tel.: 859-323-6996; Fax: 859-323-1070; E-mail: mcgong2@uky.edu.

Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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