The purpose of the study was to examine the pattern of electromyographic (EMG) activity of the rat soleus (SOL) and tibialis anterior (TA) muscles during treadmill locomotion at various speeds after 7 days of hindlimb unloading (HU). Raw EMG signals were processed to determine cycle duration, burst duration and mean EMG (burst surface divided by its duration). Cycle duration and SOL burst duration increased after HU (+7% and +5%, respectively) while TA burst duration decreased (-16%). After HU, the alternating pattern of activity between extensor and flexor muscles was maintained. Nevertheless, a co-activation of the two muscles was sometimes observed. The EMG pattern revealed no difference in the timing of the coordination between flexor and extensor muscles after HU. The delay between TA offset and SOL onset was increased (+12 ms), but this increase could be explained by the decrease in TA burst duration. Neither TA burst duration nor TA mean EMG were changed with increased treadmill speed, so that the flexor muscle activity was not related to speed of locomotion. These results would suggest that SOL activity is centrally programmed. Moreover, it is proposed that a decline in afferent feedback from SOL in rats which are suspended has an effect upon the locomotor pattern, leading to an hyperexcitability of SOL motoneurons and, via reciprocal inhibition, to a reduction in TA activity.