Blood flow (BF) to active muscle increases dramatically during exercise. This increase in BF is permitted by relaxation of smooth muscle (and ensuing vasodilation) in the vasculature of muscle tissue. Recently, attention has focused on the possible role of the endothelium-derived relaxing factor nitric oxide (EDNO) in the vasodilation of muscle vasculature during exercise. A variety of experimental approaches have been used in elucidating the role of EDNO. These include isolated vessel, isolated muscle or muscle group, and conscious exercising animal preparations. Studies utilizing isolated vessels have shown that arterioles from muscle dilate, in an endothelium-dependent manner, to stimuli present during exercise (e.g., increased flow rates). A limitation of such studies, however, is that only the potential for EDNO-induced vasodilation is indicated. The isolated muscle/muscle group approach has consistently demonstrated a role for EDNO in determining resting BF. Findings for muscle BF during contractions are equivocal. A limitation of this approach is that exercise is simulated by stimulating the motor neuron of the muscle of interest. Since this type of muscle activity elicits a relatively small active hyperemia, it may be that a role for EDNO in exercise-induced hyperemia is masked. Findings from exercising animals are equivocal. Some studies demonstrate a role for EDNO in permitting increased muscle blood flow during exercise, while others show no impact of inhibition of EDNO synthesis. Some studies suggest that the importance of EDNO may vary with the muscle (and its fiber type composition) studied. Additional research is needed to clarify the role of EDNO in mediating increased BF to skeletal muscle during exercise.