Increased resting intracellular calcium modulates NF-κB-dependent inducible nitric-oxide synthase gene expression in dystrophic mdx skeletal myotubes

Francisco Altamirano, Jose R. López, Carlos Henríquez, Tadeusz Molinski, Paul D. Allen, Enrique Jaimovich

Research output: Contribution to journalArticle

57 Scopus citations

Abstract

Duchenne muscular dystrophy (DMD) is a genetic disorder caused by dystrophin mutations, characterized by chronic inflammation and severe muscle wasting. Dystrophic muscles exhibit activated immune cell infiltrates, up-regulated inflammatory gene expression, and increased NF-κB activity, but the contribution of the skeletal muscle cell to this process has been unclear. The aim of this work was to study the pathways that contribute to the increased resting calcium ([Ca 2+] rest) observed in mdx myotubes and its possible link with up-regulation of NF-κB and pro-inflammatory gene expression in dystrophic muscle cells. [Ca 2+] rest was higher in mdx than in WT myotubes (308 ± 6 versus 113 ± 2 nM, p < 0.001). In mdx myotubes, both the inhibition of Ca 2+ entry (low Ca 2+ solution, Ca 2+-free solution, and Gd 3+) and blockade of either ryanodine receptors or inositol 1,4,5-trisphosphate receptors reduced [Ca 2+] rest. Basal activity of NF-κB was significantly up-regulated in mdx versus WT myotubes. There was an increased transcriptional activity and p65 nuclear localization, which could be reversed when [Ca 2+] rest was reduced. Levels of mRNA for TNFα, IL-1beta;, and IL-6 were similar in WT and mdx myotubes, whereas inducible nitric-oxide synthase (iNOS) expression was increased 5-fold. Reducing [Ca 2+] rest using different strategies reduced iNOS gene expression presumably as a result of decreased activation of NF-κB. We propose that NF-κB, modulated by increased [Ca 2+] rest, is constitutively active in mdx myotubes, and this mechanism can account for iNOS overexpression and the increase in reactive nitrogen species that promote damage in dystrophic skeletal muscle cells.

Original languageEnglish (US)
Pages (from-to)20876-20887
Number of pages12
JournalJournal of Biological Chemistry
Volume287
Issue number25
DOIs
StatePublished - Jun 15 2012

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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