Micropuncture techniques have been used to examine electrolyte secretion by the in vitro rabbit pancreas. The concentration profiles of the major secreted ions and digestive protein and the electrical potential profile within the pancreatic ductal system have been determined during spontaneous and secretin-stimulated secretion. The active transport of both Na and HCO(3) are the rate-controlling steps in primary secretion. Spontaneous secretion is produced primarily within the intralobular ducts. The anion composition of this primary secretion depends on the secretion rate with HCO(3) ranging from about 70 meq/liter at low rates to about 110 meq/liter at high rates. With secretin stimulation the smaller extralobular ducts also secrete and this extralobular secretion has a higher HCO(2) content than that of the intralobular secretion. In the main collecting duct the anion composition of the juice is modified further by Cl-HCO(3) exchange which appears to be a passive process depending on the average residence time of the juice in the main duct.