The factors responsible for control of glucose transport during exercise are not fully understood. We investigated the role of mechanical load in contraction-mediated glucose transport in an isolated muscle preparation. Mouse extensor digitorum longus muscles were stimulated with repeated contractions for 10 min ± N-benzyl-p-toluene sulphonamide (BTS, an inhibitor of myosin II ATPase) to block crossbridge activity. BTS inhibited force production during repeated contraction to ~5% of control. In contrast, BTS had little effect on glucose transport in the basal state (Control = 0.55±0.04; BTS = 0.47±0.09 µmol/20 min/ml) or after contraction (Control = 2.27±0.15; BTS = 2.10±0.16). BTS did not significantly alter the contraction-mediated changes in high-energy phosphates, glutathione status (a measure of oxidant status) or AMP-activated protein kinase activity. In conclusion, these data show that mechanical load plays little role in contraction-mediated glucose transport. Instead, it is likely that the increased glucose transport during contraction is a consequence of the increase in myoplasmic Ca2+ and the subsequent alterations in metabolism, e.g., increased energy turnover and production of reactive oxygen species.