The present experiments were undertaken to characterize basal release from vesicles of the regulated secretory pathway. In transfected GH3 cells, progastrin was released by the constitutive route, and mature, bioactive, amidated gastrin by the regulated secretory pathway. Studies using brefeldin A and bafilomycin A1 which inhibit progression through the Golgi complex suggested that both basal and stimulated release of amidated gastrin originated from mature secretory granules. Basal, but not stimulated, secretion of amidated gastrin was strongly inhibited at 22 degrees C. Mature secretory vesicles therefore support both basal and evoked secretion although the mechanisms underlying the two processes differ in their temperature sensitivity.