The role of proteases in excitation-contraction coupling failure in muscular dystrophy

Am J Physiol Cell Physiol. 2015 Jan 1;308(1):C33-40. doi: 10.1152/ajpcell.00267.2013. Epub 2014 Oct 8.

Abstract

Duchenne muscular dystrophy (DMD) is one of the most frequent types of muscular dystrophy. Alterations in intracellular calcium (Ca(2+)) handling are thought to contribute to the disease severity in DMD, possibly due to the activation of Ca(2+)-activated proteases. The purpose of this study was twofold: 1) to determine whether prolonged excitation-contraction (E-C) coupling disruption following repeated contractions is greater in animals lacking both dystrophin and utrophin (mdx/Utr(-/-)) compared with mice lacking only dystrophin (mdx); and 2) to assess whether protease inhibition can prevent E-C coupling failure following repeated tetani in these dystrophic mouse models. Excitation-contraction coupling was assessed using Fura-2 ratio, as an index of intracellular free Ca(2+) concentration, in response to electrical stimulation of single muscle fibers from the flexor digitorum brevis muscle. Resting Fura-2 ratio was higher in dystrophic compared with control (Con) fibers, but peak Fura-2 ratios during stimulation were similar in dystrophic and Con fibers. One hour after a series of repeated tetani, peak Fura-2 ratios were reduced by 30 ± 5.6%, 23 ± 2%, and 36 ± 3.1% in mdx, mdx/Utr(+/-), and mdx/Utr(-/-), respectively, with the greatest reduction in mdx/Utr(-/-) fibers (P < 0.05). Protease inhibition attenuated this decrease in peak Fura-2 ratio. These data indicate that E-C coupling impairment after repeated contractions is greatest in fibers lacking both dystrophin and utrophin and that prevention of protease activation can mitigate the prolonged E-C coupling impairment. These data further suggest that acute protease inhibition may be useful in reducing muscle weakness in DMD.

Keywords: Duchenne muscular dystrophy; excitation-contraction coupling; intracellular Ca2+; protease(s).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / metabolism*
  • Disease Models, Animal
  • Electric Stimulation
  • Excitation Contraction Coupling* / drug effects
  • Mice, Inbred mdx
  • Mice, Knockout
  • Muscle Fibers, Skeletal / drug effects
  • Muscle Fibers, Skeletal / enzymology*
  • Muscle Strength
  • Muscle Weakness
  • Muscle, Skeletal / drug effects
  • Muscle, Skeletal / enzymology*
  • Muscle, Skeletal / physiopathology
  • Muscular Dystrophy, Duchenne / drug therapy
  • Muscular Dystrophy, Duchenne / enzymology*
  • Muscular Dystrophy, Duchenne / genetics
  • Muscular Dystrophy, Duchenne / physiopathology
  • Peptide Hydrolases / metabolism*
  • Protease Inhibitors / pharmacology
  • Time Factors
  • Utrophin / deficiency
  • Utrophin / genetics

Substances

  • Protease Inhibitors
  • Utrophin
  • Peptide Hydrolases
  • Calcium