Ouabain exacerbates botulinum neurotoxin-induced muscle paralysis via progression of muscle atrophy in mice

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  • Fujikawa Ryu
    Department of Pathogenetic Veterinary Science, The United Graduate School of Veterinary Sciences, Gifu University
  • Muroi Yoshikage
    Department of Pathogenetic Veterinary Science, The United Graduate School of Veterinary Sciences, Gifu University Department of Basic Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine
  • Unno Toshihiro
    Department of Pathogenetic Veterinary Science, The United Graduate School of Veterinary Sciences, Gifu University The Laboratory of Pharmacology, Faculty of Applied Biological Science, Gifu University
  • Ishii Toshiaki
    Department of Pathogenetic Veterinary Science, The United Graduate School of Veterinary Sciences, Gifu University Department of Basic Veterinary Medicine, Obihiro University of Agriculture and Veterinary Medicine

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Botulinum neurotoxin serotype A (BoNT/A) inhibits acetylcholine release at the neuromuscular junction in isolated muscles, and ouabain can partially block its effect. However, it is not clear whether ouabain attenuates BoNT/A-induced neuromuscular paralysis in vivo. In this work, we investigated the effects of ouabain on BoNT/A-induced neuromuscular paralysis in mice. Ouabain was administered to mice intraperitoneally immediately after a single injection of BoNT/A into skeletal muscle. The effects of ouabain on BoNT/A-induced muscle paralysis were assessed by quantitative monitoring of muscle tension and digit abduction via the digit abduction scoring (DAS) assay. A single administration of ouabain significantly prolonged BoNT/A-induced neuromuscular paralysis. Moreover, consecutive daily injection of ouabain exacerbated BoNT/A-induced neuromuscular paralysis, and led to a significant decrease in both twitch and tetanic forces as assayed in isolated BoNT/A-injected muscles. We next looked at the effects of ouabain on BoNT/A-induced muscle atrophy. Administration of ouabain led to a decrease in the myofibrillar cross-sectional area (CSAs) by 14 post-BoNT/A injection. In addition, repeated administration of ouabain increased mRNA expression levels of ubiquitin ligases, which are markers of muscle atrophy, in BoNT/A-injected muscle. These results suggest that ouabain exacerbates BoNT/A-induced neuromuscular paralysis via a marked progression of BoNT/A-induced muscle atrophy.

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