Electrolysis stimulates creatine transport and transporter cell surface expression in incubated mouse skeletal muscle: potential role of ROS.
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 muscles of male NMRI mice (35-50 g) were incubated in an oxygenated Krebs buffer between platinum electrodes. Muscles were exposed to [(14)C]creatine for 30 min after either 12 min of repeated tetanic isometric contractions (contractions) or electrical stimulation of only the buffer before 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 (to a lesser degree) electrolysis stimulated creatine transport severalfold 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 approximately 50 and approximately 100%, respectively. The electrolysis effects on creatine uptake were completely inhibited by beta-guanidino propionic acid, 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 present results indicate that electrical field stimulation of incubated mouse muscles, independently of contractions per se, stimulates creatine transport by a mechanism that 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.
Derave
Wim
W
Dept. of Biomedical Kinesiology, Katholieke Universiteit Leuven, B-3001 Leuven, Belgium.
Straumann
Nadine
N
Olek
Robert A
RA
Hespel
Peter
P
D000818:Animals; D001693:Biological Transport, Active / Q000502:physiology; D001710:Biotin / Q000378:metabolism; D002374:Catalase / Q000378:metabolism; D003401:Creatine / Q000378:metabolism*; D004558:Electric Stimulation; D004572:Electrolysis; D006861:Hydrogen Peroxide / Q000378:metabolism; D007700:Kinetics; D008297:Male; D026901:Membrane Transport Proteins / Q000378:metabolism*; D051379:Mice; D009119:Muscle Contraction / Q000502:physiology; D018482:Muscle, Skeletal / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism
D026901:Membrane Transport Proteins; D017382:Reactive Oxygen Species; D011956:Receptors, Cell Surface; C080679:creatine transporter; D001710:Biotin; D006861:Hydrogen Peroxide; D002374:Catalase; D013482:Superoxide Dismutase; D003401:Creatine