Calpains are activated in necrotic fibers from mdx dystrophic mice.
Death of dystrophin-deficient muscle purportedly results from increases in [Ca]in that cause the activation of calpains. We have tested whether calpains play a role in this process by assaying for changes in calpain concentration and activation in peak necrotic mdx mice (4 weeks of age) and in completely regenerated mdx mice (14 weeks of age). Biochemical fractionation and immunoblotting with epitope-specific antisera allowed measurement of the concentrations of m- and mu-calpains and the extent of autoproteolytic modification. Our findings show that total calpain concentration is elevated in both 4-week and 14-week mdx mice. This increase in concentration was shown to result primarily from a significant increase in m-calpain concentration at 4 weeks. Northern analysis demonstrated that neither m- nor mu-calpain mRNA concentrations differed between mdx and controls suggesting that the increased calpain concentration results from post-translational regulation. Immunoblotting with antibodies directed against amino-terminal peptides revealed an increase in autoproteolysis of mu-calpain, indicative of increased activation. The extent of autoproteolysis of mu-calpain returns to control levels during regeneration. This is not a consequence of increased calpastatin mRNA or protein. The findings reported here support a role for calpains in both the degenerative and regenerative aspects of mdx dystrophy.
Spencer
M J
MJ
Department of Physiological Science, University of California, Los Angeles, USA.
Croall
D E
DE
Tidball
J G
JG
D000367:Age Factors; D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002154:Calpain / Q000235:genetics / Q000378:metabolism*; D016189:Dystrophin / Q000502:physiology*; D004789:Enzyme Activation; D015870:Gene Expression; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D009137:Muscular Dystrophy, Animal / Q000378:metabolism* / Q000473:pathology; D009336:Necrosis; D012333:RNA, Messenger / Q000235:genetics
D002135:Calcium-Binding Proteins; D016189:Dystrophin; D012333:RNA, Messenger; C033668:calpastatin; D002154:Calpain