The influence of venom (TSV) from the Brazilian scorpion, Tityus serrulatus, on exocrine pancreatic secretion was studied in relation to known cholinergic and peptidergic secretagogue activity. Pulse-labeling followed by chase incubation in the presence of secretagogues and various pharmacological agents revealed unique physiological characteristics of TSV in guinea pig pancreatic lobules. Exocytotic discharge of newly synthesized 3H-labeled proteins during a 3-h chase incubation showed a marked increase over basal discharge levels using logarithmic TSV doses of 0.10 to 100 micrograms/ml. This stimulation was comparable to maximal values elicited by carbachol, cholecystokinin-octapeptide (CCK-8) or caerulein and discharge kinetics were similar. TSV-mediated secretion was ATP and calcium dependent and partially inhibited by atropine. Only tetrodotoxin completely blocked TSV stimulation of newly synthesized protein discharge. Both botulinum toxin and curare had no effect on venom stimulation, indicating that TSV interaction with exocrine pancreatic cells occurs postsynaptically. Verapamil, a calcium channel antagonist, produced a moderate inhibition of TSV stimulation. When antagonists to the cholecystokinin (CCK) receptor were incubated with TSV, no change in secretory activity occurred. Therefore, TSV does not bind to CCK receptors and probably operates through its own receptor which may be an ion channel. Additionally, morphological studies in vitro revealed a high level of pancreatic secretory activity as evidenced by dense secretory acinar luminal content, reduction in zymogen granule (ZG) population, and development of exocytotic images.