Mucin plays a principal role in protecting the gastric mucosa against injury. We investigated the effect of acid secretagogues on mucin synthesis using a primary gastric mucous cell monolayer culture system of guinea pig. Significant increases in mucin synthesis were observed in response to the secretagogues pentagastrin (10(-8)M, 10(-7)M) and carbachol (10(-4)M, 10(-3)M), but not to histamine. After pretreatment with indomethacin (10(-5)M), 10(-8)M pentagastrin significantly increased mucin synthesis to 125.6 +/- 3.9%, but carbachol did not. Prostaglandin E2 release into the culture medium was significantly increased by 10(-4)M carbachol to 118.0 +/- 5.9%, but there was no change after application of pentagastrin. These findings suggest that pentagastrin and carbachol may act directly on mucous cells, and that part of the mucin synthesis-promoting action of carbachol is mediated by prostaglandins. There were no changes in intracellular cAMP concentration after the addition of these acid secretagogues. However, calcium ionophore (A23187) produced an increase in mucin synthesis, suggesting a Ca++ involvement in mucin synthesis. No differences were found in the sugar side chain structures of newly synthesized mucin glycoprotein as a result of exposure to acid secretagogues by histochemical or biochemical methods using lectins.