Normal myogenesis and increased apoptosis in myotonic dystrophy type-1 muscle cells

E. Loro, F. Rinaldi, A. Malena, E. Masiero, G. Novelli, C. Angelini, V. Romeo, M. Sandri, A. Botta, L. Vergani

Research output: Contribution to journalArticle

Abstract

Myotonic dystrophy (DM) is caused by a (CTG)(n) expansion in the 3′-untranslated region of DMPK gene. Mutant transcripts are retained in nuclear RNA foci, which sequester RNA binding proteins thereby misregulating the alternative splicing. Controversy still surrounds the pathogenesis of the DM1 muscle distress, characterized by myotonia, weakness and wasting with distal muscle atrophy. Eight primary human cell lines from adult-onset (DM1) and congenital (cDM1) patients, (CTG)(n) range 90-1800, were successfully differentiated into aneural-immature and contracting-innervated-mature myotubes. Morphological, immunohistochemical, RT-PCR and western blotting analyses of several markers of myogenesis indicated that in vitro differentiation-maturation of DM1 myotubes was comparable to age-matched controls. In all pathological muscle cells, (CTG)(n) expansions were confirmed by long PCR and RNA fluorescence in situ hybridization. Moreover, the DM1 myotubes showed the splicing alteration of insulin receptor and muscleblind-like 1 (MBNL1) genes associated with the DM1 phenotype. Considerable myotube loss and atrophy of 15-day-differentiated DM1 myotubes indicated activated catabolic pathways, as confirmed by the presence of apoptotic (caspase-3 activation, cytochrome c release, chromatin fragmentation) and autophagic (P62LC3) markers. Z-VAD treatment significantly reduced the decrease in myonuclei number and in average width in 15-day-differentiated DM1 myotubes. We thus propose that the muscle wasting typical in DM1 is due to impairment of muscle mass maintenance- regeneration, through premature apoptotic-autophagic activation, rather than altered myogenesis.

Original languageEnglish
Pages (from-to)1315-1324
Number of pages10
JournalCell Death and Differentiation
Volume17
Issue number8
DOIs
Publication statusPublished - Aug 2010

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Myotonic Dystrophy
Muscle Development
Skeletal Muscle Fibers
Muscle Cells
Apoptosis
Muscles
Myotonia
Nuclear RNA
Polymerase Chain Reaction
RNA-Binding Proteins
Muscular Atrophy
Insulin Receptor
Alternative Splicing
3' Untranslated Regions
Cytochromes c
Fluorescence In Situ Hybridization
Caspase 3
Genes
Chromatin
Atrophy

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Loro, E., Rinaldi, F., Malena, A., Masiero, E., Novelli, G., Angelini, C., ... Vergani, L. (2010). Normal myogenesis and increased apoptosis in myotonic dystrophy type-1 muscle cells. Cell Death and Differentiation, 17(8), 1315-1324. https://doi.org/10.1038/cdd.2010.33

Normal myogenesis and increased apoptosis in myotonic dystrophy type-1 muscle cells. / Loro, E.; Rinaldi, F.; Malena, A.; Masiero, E.; Novelli, G.; Angelini, C.; Romeo, V.; Sandri, M.; Botta, A.; Vergani, L.

In: Cell Death and Differentiation, Vol. 17, No. 8, 08.2010, p. 1315-1324.

Research output: Contribution to journalArticle

Loro, E, Rinaldi, F, Malena, A, Masiero, E, Novelli, G, Angelini, C, Romeo, V, Sandri, M, Botta, A & Vergani, L 2010, 'Normal myogenesis and increased apoptosis in myotonic dystrophy type-1 muscle cells', Cell Death and Differentiation, vol. 17, no. 8, pp. 1315-1324. https://doi.org/10.1038/cdd.2010.33
Loro E, Rinaldi F, Malena A, Masiero E, Novelli G, Angelini C et al. Normal myogenesis and increased apoptosis in myotonic dystrophy type-1 muscle cells. Cell Death and Differentiation. 2010 Aug;17(8):1315-1324. https://doi.org/10.1038/cdd.2010.33
Loro, E. ; Rinaldi, F. ; Malena, A. ; Masiero, E. ; Novelli, G. ; Angelini, C. ; Romeo, V. ; Sandri, M. ; Botta, A. ; Vergani, L. / Normal myogenesis and increased apoptosis in myotonic dystrophy type-1 muscle cells. In: Cell Death and Differentiation. 2010 ; Vol. 17, No. 8. pp. 1315-1324.
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