Arterial vessel responses to sodium nitroprusside (SNP) and acetylcholine (ACh) were measured in the spinotrapezius muscle of sedentary (Sed) and treadmill-trained (Tr) rats to determine whether these endothelium-dependent (ACh) and -independent (SNP) mechanisms contribute to the training-induced increase in functional vasodilation previously observed. Control and maximal vessel diameters were similar between Sed and Tr. After 8 wk of training, functional dilation (2-, 4-, and 8-Hz contractions) was enhanced in all orders of vessels studied [terminal feed artery (FA), largest arterioles (1A), and intermediate-sized arterioles (2A)], but responses to SNP were increased only in FA. Responses to ACh were not significantly increased in any vessel order. After 16 wk of training, functional dilation had regressed in Tr such that only the FA response to 4 Hz was significantly elevated relative to Sed. However, the FA and 1A responses to SNP were significantly greater in Tr than in Sed, as were the 1A and 2A responses to ACh. These results show a dissociation of functional dilation and SNP- or ACh-mediated responses, as well as age-dependent interactions, a time-dependent progression, and vessel order specificity in the adaptations to training.