Several reports have shown that the use of oral creatine (Cr) supplementation can increase performance during brief high intensity exercise in humans. The purpose of this study was to examine the separate and combined effects of Cr supplementation and high intensity run training on the performance capacity and biochemical properties of rodent skeletal muscle. Running performance was assessed following acute (10-d) and chronic (4-wk) Cr supplementation. Results indicate that Cr supplementation alone has ergogenic effects and the combination of run training plus Cr results in a more pronounced enhancement of performance than either intervention alone. The benefits of Cr supplementation were seen most clearly during repetitive bouts of high intensity interval running. Cr concentrations increased in both the slow soleus and fast plantaris muscles (P< 0.05) in response to Cr supplementation. Increased creatine concentrations appeared to be reflected in increased phosphorylated creatine (PCr). Citrate synthase (CS) activity was increased in both the soleus and plantaris muscles following training(P< 0.05). CS activity of the untrained soleus but not the plantaris responded to the dietary stimulus. There were no significant changes in either creatine phosphokinase activity or myosin heavy chain isoform distribution following training or supplementation. These results indicate that the gains in high intensity running performance seen following Cr loading are a combined result of increased aerobic (CS) and anaerobic (Cr and PCr) energy buffering capacity of the muscle.