Activation of cyclic AMP-dependent protein kinase during canine lower esophageal sphincter relaxation

MS Barnette, M Grous, TJ Torphy and HS Ormsbee

Department of Pharmacology, Smith Kline Beecham p.l.c., King of Prussia, Pennsylvania.

In isolated strips of opossum lower esophageal sphincter (LES) smooth muscle, elevation of cyclic AMP (cAMP) content is associated with relaxation. Because the activation state of cyclic nucleotide-dependent protein kinases may be a more sensitive measure of functionally important changes in cyclic nucleotide levels, we examined the ability of several pharmacological agents and electrical stimulation of the enteric neurons to activate cAMP dependent-protein kinase (cA-PK) and to relax isolated strips of LES smooth muscle. Addition of either isoproterenol or SK&F 94120, a selective inhibitor of the low Km cAMP phosphodiesterase to isolated strips of canine LES produced concentration-dependent increases in the activity ratio of cA-PK and concentration-dependent relaxations of canine LES. In contrast, although both zaprinast (M&B 22948), a selective inhibitor of the cyclic GMP selective phosphodiesterase, and sodium nitroprusside relaxed canine LES neither drug significantly increased the activity ratio of cA-PK. Furthermore, electrical stimulation of the enteric neurons produced a frequency-dependent relaxation but did not significantly activate cA-PK. To eliminate the possibility that the rapid metabolism of cAMP prevented us from observing a significant activation of cA-PK during electrical field stimulation, the ability of electrical field stimulation (1.0 Hz) to activate cA-PK was examined in the presence of 0.1 mM 3-isobutyl-1-methylxanthine. At at concentration of 0.1 mM, 3-isobutyl-1-methylxanthine, by itself, significantly increased the activity ratio of cA-PK; however, there was no additional activation produced by electrical field stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Volume 252, Issue 3, pp. 1160-1166, 03/01/1990
Copyright © 1990 by American Society for Pharmacology and Experimental Therapeutics