Rat gastric cells isolated by pronase and subdivided by Percoll into 3 fractions (F1, F2, F3) were used to study prostaglandin E2 (PGE2) formation and, as an indirect measure of parietal cell H+ production, [14C]-aminopyrine uptake. Cells that had not been fractionated, with 20 to 25% parietal cells, contained at 0 degree C 1.7 +/- 0.35 (s.e.mean) ng PGE2 10(8) cells-1. During incubation at 37 degrees C these cells steadily synthesized up to 4.36 +/- 0.73 ng PGE2 10(8) cells-1 from endogenous substrate. Indomethacin in concentrations higher than 10(-6) mol 1(-1) inhibited this basal formation completely, but 10(-4) mol 1(-1) did not reduce the cellular PGE2 level below 1.4 +/- 0.2 ng 10(8) cells-1. Arachidonic acid in concentrations higher than 10(-5) mol 1(-1) evoked an abundant formation of PGE2, and 10(-4) mol 1(-1) built up a plateau of over 7.5 +/- 1.65 ng PGE2 10(8) cells-1 within 15 min. PGE2 formation in cell fractions increased significantly with the number of parietal cells per assay tube. Indomethacin (10(-8) to 10(-4) mol 1(-1] did not influence the histamine-stimulated uptake of [14C]-aminopyrine, while arachidonic acid (10(-5) to 10(-4) mol 1(-1] inhibited this process. PGE2 formation in response to arachidonic acid was prevented by indomethacin, but the inhibition of aminopyrine uptake by arachidonic acid could not be prevented by indomethacin. The data suggest that isolated gastric cells of the rat sustain constant PGE2 synthesis in vitro, which is more pronounced in parietal than in mucosal and chief cells. PGE2 may exert different effects within distinct gastric cell types.