Catabolism of Cytoplasmic and Intramitochondrial Adenine Nucleotides in C2C12 Skeletal Myotube under Chemical Hypoxia.

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  • MATSUKI Naoaki
    Department of Veterinary Clinical Pathobiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo
  • INABA Mutsumi
    Department of Veterinary Clinical Pathobiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo
  • ONO Kenichiro
    Department of Veterinary Clinical Pathobiology, Graduate School of Agricultural and Life Sciences, The University of Tokyo

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Loss of adenosine-5'-triphosphate (ATP) and accumulation of inosine-5'-monophosphate (IMP) are the major purine metabolic changes in the skeletal muscle during hypoxia. This study addressed whether chemical metabolic inhibition reflects those changes in cultured skeletal myotube. For this aim, mouse-derived C2C 12 myotubes were cultured in Hank's balanced saline solution containing 2 mM sodium cyanide (CN) and/or 1 mM iodoacetic acid (IAA) up to 180 min. Inhibition of oxidative phosphorylation by CN induced a minimal change in the intracellular adenine nucleotide levels during 180 min. Blockage of glycolysis with IAA caused an over 90% decrease in adenine nucleotides both in the cytoplasmic and intramitochondrial spaces, accompanied with allantoin release. Since 1 mM allopurinol entirely inhibited the allantoin generation, xanthine dehydrogenase/oxidase was found to play a key role in the purine catabolism in IAA-treated C2C12 myotubes. By the combined treatment with CN+IAA, ATP exhaustion and IMP accumulation was achieved with significant cell injury. These changes were comparable with those in skeletal muscles during hypoxia, indicating that our model with CN+IAA is well applicable to the investigation of hypoxia-induced myopathy.

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