Electrical field stimulation of isolated, incubated rodent skeletal muscles is a frequently used model to study the effects of contractions on muscle metabolism. In this study, this model was used to investigate the effects of electrically-stimulated contractions on creatine transport. Soleus and extensor digitorum longus (EDL) muscles of male NMRI mice (35-50g) were incubated in an oxygenated Krebs-buffer between platinum electrodes. Muscles were exposed to ¹⁴C-labelled creatine for 30 min following either 12 min of repeated tetanic isometric contractions (contractions) or following electrical stimulation of buffer only prior to incubation of the muscle (electrolysis). Electrolysis was also investigated in the presence of the reactive oxygen species (ROS) scavenging enzymes superoxide dismutase (SOD) and catalase. Both contractions and electrolysis (to a lesser degree) stimulated creatine transport several fold over basal. The amount of electrolysis, but not contractile activity, induced (determined) creatine transport stimulation. Incubation with SOD and catalase at 100 and 200 U/ml decreased electrolysis-induced creatine transport by ~50% and ~100%, respectively. The electrolysis effects on creatine uptake were completely inhibited by GPA, a competitive inhibitor of (creatine for) the creatine transporter (CRT), and were accompanied by increased cell surface expression of CRT. Muscle glucose transport was not affected by electrolysis. The current results indicate that electrical field stimulation of incubated mouse muscles, independent of contractions per se, stimulates creatine transport by a mechanism which depends on electrolysis-induced formation of ROS in the incubation buffer. The increased creatine uptake is paralleled by an increased cell surface expression of the creatine transporter.