To provide more comprehensive information on the extent and pattern of muscle activation during running, we determined lower extremity muscle activation by using exercise-induced contrast shifts in magnetic resonance (MR) images during horizontal and uphill high-intensity (115% of peak oxygen uptake) running to exhaustion (2.0-3.9 min) in 12 young women. The mean percentage of muscle volume activated in the right lower extremity was significantly (P <0.05) greater during uphill (73 +/- 7%) than during horizontal (67 +/- 8%) running. The percentage of 13 individual muscles or groups activated varied from 41 to 90% during horizontal running and from 44 to 83% during uphill running. During horizontal running, the muscles or groups most activated were the adductors (90 +/- 5%), semitendinosus (86 +/- 13%), gracilis (76 +/- 20%), biceps femoris (76 +/- 12%), and semimembranosus (75 +/- 12%). During uphill running, the muscles most activated were the adductors (83 +/- 8%), biceps femoris (79 +/- 7%), gluteal group (79 +/- 11%), gastrocnemius (76 +/- 15%), and vastus group (75 +/- 13%). Compared with horizontal running, uphill running required considerably greater activation of the vastus group (23%) and soleus (14%) and less activation of the rectus femoris (29%), gracilis (18%), and semitendinosus (17%). We conclude that during high-intensity horizontal and uphill running to exhaustion, lasting 2-3 min, muscles of the lower extremity are not maximally activated, suggesting there is a limit to the extent to which additional muscle mass recruitment can be utilized to meet the demand for force and energy. Greater total muscle activation during exhaustive uphill than during horizontal running is achieved through an altered pattern of muscle activation that involves increased use of some muscles and less use of others.