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  • Title: Effects of phorbol ester treatment on dibutyryl cyclic adenosine-5' monophosphate- and carbachol-stimulated aminopyrine accumulation in isolated rat parietal cells.
    Author: Kopp R, Pfeiffer A.
    Journal: Scand J Gastroenterol; 2000 Jul; 35(7):686-93. PubMed ID: 10972170.
    Abstract:
    BACKGROUND: The functional role of the intracellular diacylglycerol/protein kinase C second-messenger pathway in the regulation of gastric acid secretion and the effects on the involved inositotrisphosphate/Ca2+/calmodulin system are not well understood, and contradictory data have been reported. We therefore evaluated the effects of phorbol ester treatment (tetradecanoylphorbol-12,13-acetate (TPA)) on dibutyryl cyclic adenosine-5' monophosphate (dBcAMP)- and carbachol-stimulated aminopyrine (AP) accumulation in comparison with intracellular alterations of the phospholipase C/inostol phosphate signal transduction pathway in isolated rat gastric parietal cells. METHODS: [14C]AP accumulation was determined as an indirect measure of gastric acid secretion. Inositolphosphate second-messenger activation was investigated with [3H]inositolmonophosphate release in [3H]-myoinositol prelabeled rat gastric parietal cells. RESULTS: TPA at a low concentration of 5 nM caused a small (45%) but significant increase in carbachol (0.1 mM)-stimulated AP accumulation, which was dose-dependently inhibited by higher concentrations of TPA with corresponding shifts in the dose-response curve for carbachol-stimulated AP accumulation. AP uptake stimulated by dBcAMP (0.1 mM) and the synergistic stimulatory effect induced by carbachol together with dBcAMP were inhibited by TPA at all concentrations investigated. In the presence of increasing concentrations of the calcium ionophore ionomycin (10(-8)-10(-5) M) TPA at 5 nM increased AP accumulation (AP ratio was 4.02 with 5 nM TPA versus 1.23 in the absence of TPA; P < 0.05), indicating that phorbol ester stimulates AP uptake in rat parietal cells. Simultaneous investigation of [14C]AP accumulation and [3H]inositol monophosphate release showed that inhibitory effects of TPA on carbachol- and carbachol plus dBcAMP-stimulated cells are mediated by an inhibition of the receptor/G-protein/phospholipase C interaction, leading to a reduction of inositolphosphate release. The costimulation of rat parietal cells with dBcAMP, ionomycin, and TPA (5 nM) did not reproduce the synergistic effects of carbachol together with dBcAMP on AP accumulation, suggesting that carbachol-stimulated AP uptake seems to be additionally mediated by a still unknown pathway independent of intracellular calcium release or protein kinase C activation.
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