Effect of cyclopiazonic acid, an inhibitor of the sarcoplasmic reticulum Ca-ATPase, on skeletal muscles from normal and mdx mice

Acta Physiol Scand. 2005 Jul;184(3):173-86. doi: 10.1111/j.1365-201X.2005.01450.x.

Abstract

Aim: In this study, we investigated Ca2+ loading by the sarcoplasmic reticulum in skeletal muscle from mdx mice, an animal model of human Duchenne's muscular dystrophy, at two stages of development: 4 and 11 weeks.

Method: Experiments were conducted on fast- (extensor digitorum longus, EDL) and slow- (soleus) twitch muscles expressing different isoforms of Ca2+-ATPase, which is responsible for the uptake of Ca2+ by the sarcoplasmic reticulum.

Results: In sarcoplasmic reticulum vesicles, the ATP-dependent activity and sensitivity to cyclopiazonic acid (CPA), an inhibitor of the sarcoplasmic reticulum Ca2+-ATPase, were similar in mdx and normal EDL muscle. Furthermore, in chemically-skinned fibres from both normal and mdx muscles, the presence of CPA induced a decrease in Ca2+ uptake by the sarcoplasmic reticulum. However, the sensitivity to CPA was lower in mdx EDL muscle than in normal muscle. In addition, in EDL muscle from 4-week-old mdx mice, the expression of the slow Ca2+-pump isoform (SERCA2a) was significantly increased, without any accompanying change in slow myosin expression. In contrast, the expression and function of the Ca2+-ATPase in mdx soleus muscles at 4- and 11-weeks of development did not differ from those in age-matched controls.

Conclusion: These findings show that in dystrophic muscle, where the Ca2+ homeostasis was perturbed, the Ca2+ handling by the sarcoplasmic reticulum was altered in fast-twitch muscle, and this was associated with the expression of the slow isoform of SERCA. In these muscles, reduced Ca2+ uptake could then contribute to an elevated concentration of Ca2+ in the cytosol, and also to Ca2+ depletion of the sarcoplasmic reticulum.

MeSH terms

  • Animals
  • Calcium / pharmacokinetics
  • Calcium-Transporting ATPases / analysis
  • Calcium-Transporting ATPases / antagonists & inhibitors*
  • Enzyme Inhibitors / pharmacology*
  • Immunohistochemistry / methods
  • Indoles / pharmacology*
  • Male
  • Mice
  • Mice, Inbred mdx
  • Muscle Contraction / physiology
  • Muscle Proteins / metabolism
  • Muscle, Skeletal / drug effects*
  • Muscular Dystrophy, Duchenne / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Sarcoplasmic Reticulum / enzymology
  • Sarcoplasmic Reticulum / metabolism*
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases

Substances

  • Enzyme Inhibitors
  • Indoles
  • Muscle Proteins
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases
  • Calcium-Transporting ATPases
  • Calcium
  • cyclopiazonic acid