Lipoprotein lipase activity was measured in the three skeletal muscle fiber types of untrained rats and in those of rats subjected to a 12-wk program of treadmill running. Lipoprotein lipase activity in slow-twitch red fibers was approximately 14- to 20-fold higher (P less than 0.001) than that in fast-twitch white and approximately 2-fold higher (P less than 0.001) than that in fast-twitch red fibers in the untrained animals. These results suggest that, in sedentary animals, mainly slow-twitch red and fast-twitch red fibers are capable of taking up plasma triglyceride fatty acids. Regularly performed endurance exercise resulted in significant increase (2- to 4.5-fold) in lipoprotein lipase activity in the three muscle fiber types examined. The increase in lipoprotein lipase activity in response to treadmill running suggests that exercise increases the capacity of these fibers to take up and oxidize plasma triglyceride fatty acids. Cardiac muscle did not undergo an exercise-induced increase in the levels of activity of lipoprotein lipase similar to that seen in skeletal muscle.