To investigate the role of insulin in the potentiation effect of secretin and cholecystokinin (CCK) on pancreatic exocrine secretion, the pancreas was isolated from rats and perfused with modified Krebs-Henseleit solution containing glucose at three concentrations. Intraarterial glucose at concentrations of 2.5, 10, and 25 mMproduced modest but significant increases in both the pancreatic flow rate and the amylase output in a concentration-dependent manner. The mixture of secretin and CCK at concentrations of 18.5 and 14 pM, respectively, added to the glucose solutions augmented the pancreatic flow rate and amylase output in relation to the glucose concentration. In the streptozotocin-treated pancreas, the mixture of secretin and CCK failed to augment the pancreatic exocrine secretion unless exogenous insulin was added to the perfusate. Secretin markedly potentiated the CCK-induced amylase output when insulin was present in the circulation. However, CCK did not potentiate the secretin-induced flow rate even if insulin was present in the circulation. Insulin did not affect the actions of secretin alone but it potentiated the actions of CCK alone in both the pancreatic flow rate and the amylase output. It is concluded from the above results that insulin intensifies the combined actions of secretin and CCK in pancreatic exocrine secretion by potentiating the CCK action. Furthermore, in the presence of insulin, secretin is able to potentiate the pancreatic enzyme secretion stimulated by CCK.