Secretagogue-induced [14C]aminopyrine uptake in isolated equine parietal cells

Equine oxyntic mucosal cells were obtained by sequential exposure to pronase and collagenase. Acid production by parietal cells was assessed by uptake of [14C]aminopyrine (AP), a weak base that accumulates in intracellular acidic spaces. Incubation for various times revealed a maximal AP uptake at 10 minutes for histamine and carbachol. Similar secretagogue responses were observed for parietal cells from the mucosal cell preparation or after enrichment by elutriation. Histamine and isobutyl-methylxanthine (IBMX) stimulated AP uptake with a dose-dependent response and maximal effective concentration of 100 microM. Carbachol, 1 to 100 microM, and pentagastrin (PG), 1 to 1,000 nM, were ineffective stimulants of AP uptake. The AP uptake values for 100 microM IBMX, 1 microM carbachol, or 100 nM PG were 77 +/- 6%, 50 +/- 3.2%, and 40 +/- 4.5%, respectively, of that observed with maximal stimulation by 100 microM histamine (mean +/- SEM, n = 4 to 14). Uptake of AP by nonstimulated control cells was 36 +/- 3.6% of maximal histamine stimulation. The AP accumulations during control conditions and after stimulation with 100 microM histamine and IBMX, 1 microM carbachol, or 100 nM PG were 1.18 +/- 0.39, 2.81 +/- 0.85, 1.93 +/- 0.48, 1.44 +/- 0.36, and 1.23 +/- 0.33 pmol of AP/10(5) parietal cells, respectively. Individual histamine dose-response curves were shifted to the right by increasing ranitidine and cimetidine concentrations (0.1 to 50 microM). These results indicate that isolated equine parietal cells are maximally stimulated by histamine and minimally stimulated by carbachol and PG.