Directional blood flow in panreatic islets may be important for regulation of islet hormone release. We therefore perfused an isolated canine pancreas via the celiac artery (arterial perfusion) and then via the portal vein (venous perfusion) in the same pancreas. Basal insulin and glucagon levels and their rate of release in response to 10 mM arginine, 11 mM glucose, 500 pg/ml somatostatin, or 500 pg/ml glucagon were similar under both conditions. However, the inhibition of glucagon release due to somatostatin and its recovery after the cessation of somatostatin infusion was poor in the case of venous perfusion. The basal somatostatin level and its release in response to 10 mM arginine, 11 mM glucose, and 500 pg/ml glucagon during venous perfusion was significantly higher than that during arterial perfusion. From these results, it is speculated that the directional blood flow in pancreatic islets may not be essential for regulation of hormone release from the canine pancreas or that such directionality does not exist in canine pancreatic islets, and that a considerable portion of released somatostatin may be taken up by the pancreas located downstream—probably in the exocrine pancreas.