The changes seen in the T1 and T2 relaxation times, water content and size of the extracellular fluid spaces of rat muscle samples following 15 days of denervation were studied by in vitro proton NMR spectrometry (10 MHz). Two different skeletal muscle groups (gastrocnemius and soleus) were studied. Denervation led to longer T1 values: 548 +/- 61 msec vs. 486 +/- 16 msec (P less than .05) for the gastrocnemius and 581 +/- 27 msec vs. 521 +/- 25 msec (P less than .05) for the soleus. Similar increases in T2 were measured. The sizes of the extracellular fluid spaces of denervated muscle were significantly larger despite a minor increase in total water content. Overall, the relaxation times of skeletal muscle correlated better with the size of the extracellular fluid space than with the total water content.