Cast immobilization causes skeletal muscle disuse atrophy and an increased susceptibility to muscle damage. The objective of this study was to explore the utility of noninvasive magnetic resonance (MR) imaging to monitor muscle damage in the lower hindlimb muscles of the mouse during reloading following cast immobilization and to compare the findings in different muscles. The hindlimbs of C57BL6 mice were immobilized for 2 weeks in plantarflexion using a bilateral casting model. Following immobilization the mice were allowed to reambulate and muscle damage was monitored at different times. Cage-restricted reloading following cast immobilization induced a significant shift (P < 0.0001) in the transverse (T2) relaxation characteristics of the postural slow-twitch soleus muscle, but not in the neighboring gastrocnemius. Soleus T2 values peaked at 2 days of reloading. Muscle-specific changes in MR T2 relaxation properties correlated with uptake of Evans blue dye, a histological marker of muscle damage. This study demonstrates that T2 MR imaging can be implemented to monitor noninvasively and sequentially muscle-specific damage during reloading following limb disuse.