A method for portal venous, aortic, and gastric cannulation in the rat is described. Changes in mean portal venous and aortic blood glucose and lactate concentrations after an intragastric infusion of d-glucose to chronically catheterized rats (after regaining preoperative weight) were compared to those of acutely catheterized rats (1 h after catheter placement). Both portal venous and aortic catheters were patent for blood sampling in 67% of rats 8 days after catheter placement and 42% after 14 days. After intragastric infusion of 10 ml of 0.29 M dextrose, mean aortic blood glucose and lactate concentrations in chronically catheterized rats increased 80 and 73%, respectively, 15 min after infusion and decreased to preinfusion concentrations by 60 min. In acutely catheterized rats, mean aortic glucose and lactate concentrations increased linearly for 60 min. The concentration differences between portal venous and aortic blood glucose and lactate concentrations in chronically catheterized rats were not significantly different than baseline differences. The portal venous glucose concentrations were significantly greater than aortic concentrations in acutely catheterized rats 15, 30, and 45 min after the glucose infusion when compared to baseline differences. These data suggest that the rate of glucose uptake and metabolism to lactate is significantly altered in acutely catheterized rats when compared to chronically catheterized animals.