The effect of depletion of serotonin stores on vagally stimulated gastric acid secretion and motility was studied in rats. Pretreatment of rats with parachlorophenylalanine (p-CPA) produced a 57% reduction in the intraluminal gastric release of serotonin and a 43-100% potentiation of the gastric acid secretory response elicited by intracisternal injection of the stable TRH analogue RX 77368 in conscious pylorusligated rats or in urethane-anesthetized rats with an acute gastric fistula. p-CPA also enhanced the vagally stimulated gastric acid output produced by intravenous injection of baclofen. In contrast, p-CPA pretreament had no effect on gastric acid secretion stimulated by bethanechol, histamine, or pentagastrin. Selective depletion of central serotonin stores by the neurotoxin 5,7-dihydroxytryptamine (5,7-DHT) given alone or combined with parachloroamphetamine pretreatment did not alter RX 77368-stimulated gastric acid secretion. In addition, gastric contractility stimulated by intracisternal injection of RX 77368 was significantly enhanced by p-CPA but not by 5,7-DHT pretreatment; whereas the contractile response to carbachol was not altered by p-CPA pretreatment. These results suggest that depletion of peripheral but not central serotonergic stores potentiates gastric acid secretion and contractility stimulated by vagally, but not peripherally, acting gastric stimulants. Thus, peripheral serotonin may exert an inhibitory tone on vagally stimulated gastric acid secretion and motility in the rat.