ABSTRACT-In the intestinal smooth muscle of guinea pig taenia caeci, acetylcholine and carbachol induced a transient contraction followed by a sustained contraction. The magnitudes of the transient and sustained contractions were similar when muscle was stimulated with acetylcholine (0.1 uM -1 mM) or a lower concentration (0.1 rM) of carbachol. However, higher concentrations of carbachol (1-100 pM) induced significantly smaller sustained contraction than the transient contraction. In the 45 mM KCl-stimulated strips, addition of 100 pM carbachol induced a transient increase followed by a sustained decrease in the contractile tension. In contrast, acetylcholine (0.1 pM -1 mM) showed only weak inhibitory effects on the high K+-induced contraction either in the absence or presence of a cholinesterase inhibitor, 0.5 pM diisopropylfluorophosphate.The same concentration of diisopropylfluorophosphate shifted the concentration-response curve for acetylcholine to lower concentrations. In the muscles pretreated with 3 pM phorbol 12-myristate 13-acetate for 24 hr to desensitize protein kinase C, sustained contractions induced by higher concentrations of carbachol (1-100 pM) were significantly greater than those in the strips without the treatment with phorbol ester. However, the transient contraction and the contraction induced by a lower concentration (0.1 I-pM) of carbachol were not changed by the treatment with phorbol ester. Pretreatment with phorbol ester attenuated the inhibitory effect of carbachol on the high K+-induced contraction. These results suggest that the inhibitory effects of carbachol is composed of two phases: protein kinase Cindependent transient inhibition and protein kinase C-dependent sustained inhibition.Keywords: Smooth muscle (intestinal), Acetylcholine, Carbachol, Phorbol ester, Protein kinase C It has been shown that acetylcholine and carbachol induce contraction by increasing the Ca2+ influx in intestinal smooth muscle (1). Higher concentrations of acetylcholine and carbachol also stimulate phosphoinositide turnover (2-4), and one of the hydrolysis products, inositol 1,4,5-trisphosphate, releases Ca2+ to induce transient contraction (5).We have previously reported that high concentrations of carbachol inhibit the sustained contraction induced by carbachol itself by a decrease of cytosolic Ca2+ in the intestinal smooth muscle of guinea pig taenia caeci. We have also reported that high concentrations of carbachol inhibit high K+ induced contraction (6). Phorbol ester, 12-deoxyphorbol 13-isobutyrate, also inhibited high K+-induced contraction, and this effect was abolished when the protein kinase C was desensitized (7). These results suggest that the inhibitory effect of a high concentration of carbachol on high K+-induced contraction may be Male guinea pigs, weighing 250-300 g, were killed by a blow on the neck and bled. A section of taenia (5 -10 mm in length) was dissected from the caecum. For the desensitization of protein kinase C, the muscle strips were incubated in Dulbecco's modified Eagle ...