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Am J Physiol Gastrointest Liver Physiol 283: G282-G291, 2002; doi:10.1152/ajpgi.00357.2001
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Vol. 283, Issue 2, G282-G291, August 2002

PGF2alpha -induced contraction of cat esophageal and lower esophageal sphincter circular smooth muscle

Weibiao Cao, Karen M. Harnett, Jose Behar, and Piero Biancani

Department of Medicine, Rhode Island Hospital and Brown University School of Medicine, Providence, Rhode Island 02903

Lower esophageal sphincter (LES) tone depends on PGF2alpha and thromboxane A2 acting on receptors linked to Gi3 and Gq to activate phospholipases and produce second messengers resulting in muscle contraction. We therefore examined PGF2alpha signal transduction in circular smooth muscle cells isolated by enzymatic digestion from cat esophagus (Eso) and LES. In Eso, PGF2alpha -induced contraction was inhibited by antibodies against the alpha -subunit of G13 and the monomeric G proteins RhoA and ADP-ribosylation factor (ARF)1 and by the C3 exoenzyme of Clostridium botulinum. A [35S]GTPgamma S-binding assay confirmed that G13, RhoA, and ARF1 were activated by PGF2alpha . Contraction of Eso was reduced by propranolol, a phospholipase D (PLD) pathway inhibitor and by chelerythrine, a PKC inhibitor. In LES, PGF2alpha -induced contraction was inhibited by antibodies against the alpha -subunit of Gq and Gi3, and a [35S]GTPgamma S-binding assay confirmed that Gq and Gi3 were activated by PGF2alpha . PGF2alpha -induced contraction of LES was reduced by U-73122 and D609 and unaffected by propranolol. At low PGF2alpha concentration, contraction was blocked by chelerythrine, whereas at high concentration, contraction was blocked by chelerythrine and CGS9343B. Thus, in Eso, PGF2alpha activates a PLD- and protein kinase C (PKC)-dependent pathway through G13, RhoA, and ARF1. In LES, PGF2alpha receptors are coupled to Gq and Gi3, activating phosphatidylinositol- and phosphatidylcholine-specific phospholipase C. At low concentrations, PGF2alpha activates PKC. At high concentration, it activates both a PKC- and a calmodulin-dependent pathway.

smooth muscle contraction; prostaglandins; G proteins; phospholipases


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