Separating myoblast differentiation from muscle cell fusion using IGF-I and the p38 MAP kinase inhibitor SB202190

Samantha Gardner, Sean M. Gross, Larry David, John E. Klimek, Peter Rotwein

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

The p38 MAP kinases play critical roles in skeletal muscle biology, but the specific processes regulated by these kinases remain poorly defined. Here we find that activity of p38α/β is important not only in early phases of myoblast differentiation, but also in later stages of myocyte fusion and myofibrillogenesis. By treatment of C2 myoblasts with the promyogenic growth factor insulin-like growth factor (IGF)-I, the early block in differentiation imposed by the p38 chemical inhibitor SB202190 could be overcome. Yet, under these conditions, IGF-I could not prevent the later impairment of muscle cell fusion, as marked by the nearly complete absence of multinucleated myofibers. Removal of SB202190 from the medium of differentiating myoblasts reversed the fusion block, as multinucleated myofibers were detected several hours later and reached ∼90% of the culture within 30 h. Analysis by quantitative mass spectroscopy of proteins that changed in abundance following removal of the inhibitor revealed a cohort of upregulated muscle-enriched molecules that may be important for both myofibrillogenesis and fusion. We have thus developed a model system that allows separation of myoblast differentiation from muscle cell fusion and should be useful in identifying specific steps regulated by p38 MAP kinase-mediated signaling in myogenesis.

Original languageEnglish (US)
Pages (from-to)C491-C500
JournalAmerican Journal of Physiology - Cell Physiology
Volume309
Issue number7
DOIs
StatePublished - Sep 1 2015

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Cell Fusion
Myoblasts
p38 Mitogen-Activated Protein Kinases
Insulin-Like Growth Factor I
Muscle Cells
Muscle Development
Mass Spectrometry
Intercellular Signaling Peptides and Proteins
Skeletal Muscle
Phosphotransferases
Muscles
4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)imidazole
Proteins

Keywords

  • IGF-I signaling
  • Muscle cell fusion
  • Muscle differentiation
  • Myofiber formation
  • P38

ASJC Scopus subject areas

  • Cell Biology
  • Physiology

Cite this

Separating myoblast differentiation from muscle cell fusion using IGF-I and the p38 MAP kinase inhibitor SB202190. / Gardner, Samantha; Gross, Sean M.; David, Larry; Klimek, John E.; Rotwein, Peter.

In: American Journal of Physiology - Cell Physiology, Vol. 309, No. 7, 01.09.2015, p. C491-C500.

Research output: Contribution to journalArticle

Gardner, Samantha ; Gross, Sean M. ; David, Larry ; Klimek, John E. ; Rotwein, Peter. / Separating myoblast differentiation from muscle cell fusion using IGF-I and the p38 MAP kinase inhibitor SB202190. In: American Journal of Physiology - Cell Physiology. 2015 ; Vol. 309, No. 7. pp. C491-C500.
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