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Vol. 289, Issue 2, 632-640, May 1999

Differential Induction of Nitric Oxide Synthase in Rat Gastric and Vascular Smooth Muscle Tissue: Distinct Tissue Distribution and Distinctive Signaling Pathways1

Xi-Long Zheng , Yu Gui , Keith A. Sharkey and Morley D. Hollenberg

Endocrine (X.-L.Z., Y.G., M.D.H.) and Neuroscience (K.A.S.) Research Groups, Departments of Physiology (K.A.S.), Pharmacology and Therapeutics (X.-L.Z., Y.G., M.D.H.), and Medicine (X.-L.Z., Y.G., M.D.H.), The University of Calgary, Faculty of Medicine, Calgary, Alberta, Canada

In rat aorta rings (RA) and in gastric circular muscle (CM) and gastric longitudinal muscle (LM) preparations maintained in vitro, inducible nitric oxide synthase (iNOS) induction was monitored functionally (1 mM L-arginine-induced relaxation), biochemically (appearance of iNOS mRNA), and immunohistochemically. Functional iNOS (L-arginine-mediated relaxation) was induced in RA and CM tissues (but NOT in the LM preparation) over 2 to 5 h. iNOS induction was detected by immunocytochemistry in RA smooth muscle elements and in macrophage-like cells in CM. Functional iNOS induction correlated with iNOS mRNA induction. In the RA and CM, functional iNOS induction was blocked by both actinomycin D and cycloheximide; actinomycin D also blocked the appearance of iNOS mRNA in both tissues. In contrast, cycloheximide blocked CM (but not RA) iNOS mRNA induction. In CM tissue, functional iNOS induction was not affected by genistein, tyrphostin 47/AG213, or vanadate. But, in the RA, both genistein and tyrphostin 47/AG213 blocked the appearance of functional iNOS; neither inhibitor prevented the appearance of RA iNOS mRNA. Vanadate, in the RA tissue, blocked both the appearance of iNOS mRNA and the induction of functional iNOS. In RA tissue, but not in the CM, inhibitors of NF-kappa B activation blocked the appearance of both functional iNOS and iNOS mRNA. We conclude that in different smooth muscle preparations (aorta versus gastric), there can be a differential induction of iNOS mRNA and "functional" iNOS not only in different cellular elements but also in terms of different signaling pathways.

0022-3565/99/2892-0632$03.00/0
THE JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS
Copyright © 1999 by The American Society for Pharmacology and Experimental Therapeutics


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