Causes of excitation-induced muscle cell damage in isometric contractions: mechanical stress or calcium overload?
Prolonged or unaccustomed exercise leads to muscle cell membrane damage, detectable as release of the intracellular enzyme lactic acid dehydrogenase (LDH). This is correlated to excitation-induced influx of Ca2+, but it cannot be excluded that mechanical stress contributes to the damage. We here explore this question using N-benzyl-p-toluene sulfonamide (BTS), which specifically blocks muscle contraction. Extensor digitorum longus muscles were prepared from 4-wk-old rats and mounted on holders for isometric contractions. Muscles were stimulated intermittently at 40 Hz for 15-60 min or exposed to the Ca2+ ionophore A23187. Electrical stimulation increased 45Ca influx 3-5 fold. This was followed by a progressive release of LDH, which was correlated to the influx of Ca2+. BTS (50 microM) caused a 90% inhibition of contractile force but had no effect on the excitation-induced 45Ca influx. After stimulation, ATP and creatine phosphate levels were higher in BTS-treated muscles, most likely due to the cessation of ATP-utilization for cross-bridge cycling, indicating a better energy status of these muscles. No release of LDH was observed in BTS-treated muscles. However, when exposed to anoxia, electrical stimulation caused a marked increase in LDH release that was not suppressed by BTS but associated with a decrease in the content of ATP. Dynamic passive stretching caused no increase in muscle Ca2+ content and only a minor release of LDH, whereas treatment with A23187 markedly increased LDH release both in control and BTS-treated muscles. In conclusion, after isometric contractions, muscle cell membrane damage depends on Ca2+ influx and energy status and not on mechanical stress.
Fredsted
Anne
A
Department of Physiology and Biophysics, University of Aarhus, Denmark. af@fi.au.dk
Gissel
Hanne
H
Madsen
Klavs
K
Clausen
Torben
T
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D012090:Cumulative Trauma Disorders / Q000209:etiology* / Q000503:physiopathology*; D004558:Electric Stimulation / Q000009:adverse effects*; D005260:Female; D066298:In Vitro Techniques; D007537:Isometric Contraction; D008297:Male; D018482:Muscle, Skeletal / Q000293:injuries* / Q000503:physiopathology*; D051381:Rats; D017208:Rats, Wistar; D013314:Stress, Mechanical
D000255:Adenosine Triphosphate; D002118:Calcium