Inhibition of Drp1-dependent mitochondrial division impairs myogenic differentiation

Boa Kim, Ji Seok Kim, Yisang Yoon, Mayra C. Santiago, Michael D. Brown, Joon Young Park

Research output: Contribution to journalArticle

40 Citations (Scopus)

Abstract

Mitochondria are dynamic organelles forming a tubular network that is continuously fusing and dividing to control their morphology and functions. Recent literature has shed new light on a potential link between the dynamic behavior of mitochondria and muscle development. In this study, we investigate the role of mitochondrial fission factor dynamin-related protein 1 (Drp1) in myogenic differentiation. We found that differentiation of C2C12 myoblasts induced by serum starvation was accompanied by a gradual increase in Drp1 protein expression (to ~350% up to 3 days) and a fast reduction of Drp1 phosphorylation at Ser-637 (to ~30%) resulting in translocation of Drp1 protein from the cytosol to mitochondria. During differentiation, treatment of myoblasts with mitochondrial division inhibitor (mdivi-1), a specific inhibitor of Drp1 GTPase activity, caused extensive formation of elongated mitochondria, which coincided with increased apoptosis evidenced by both enhanced caspase-3 activity and increased number of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive cells. Furthermore, the mdivi-1 -treated myotubes (day 3 in differentiation media) showed a reduction in mitochondrial DNA content, mitochondrial mass, and membrane potential in a dose-dependent manner indicating defects in mitochondrial biogenesis during myogenic differentiation. Most interestingly, mdivi-1 treatment significantly suppressed myotube formation in both C2C12 cells and primary myoblasts. Likewise, stable overexpression of a dominant negative mutant Drp1 (K38A) dramatically reduced myogenic differentiation. These data suggest that Drp-1-dependent mitochondrial division is a necessary step for successful myogenic differentiation, and perturbation of mitochondrial dynamics hinders normal mitochondrial adaptations during muscle development. Therefore, in the present study, we report a novel physiological role of mitochondrial dynamics in myogenic differentiation.

Original languageEnglish (US)
Pages (from-to)R927-R938
JournalAmerican Journal of Physiology - Regulatory Integrative and Comparative Physiology
Volume305
Issue number8
DOIs
StatePublished - Oct 15 2013

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Dynamins
Mitochondrial Dynamics
Myoblasts
Mitochondria
Proteins
Muscle Development
Skeletal Muscle Fibers
DNA Nucleotidylexotransferase
Mitochondrial Membrane Potential
GTP Phosphohydrolases
Organelle Biogenesis
Starvation
Mitochondrial DNA
Caspase 3
Organelles
Cytosol
Phosphorylation
Apoptosis
Serum

Keywords

  • Dynamin-related protein 1
  • Mdivi-1
  • Mitochondrial dynamics
  • Myogenesis

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)

Cite this

Inhibition of Drp1-dependent mitochondrial division impairs myogenic differentiation. / Kim, Boa; Kim, Ji Seok; Yoon, Yisang; Santiago, Mayra C.; Brown, Michael D.; Park, Joon Young.

In: American Journal of Physiology - Regulatory Integrative and Comparative Physiology, Vol. 305, No. 8, 15.10.2013, p. R927-R938.

Research output: Contribution to journalArticle

Kim, Boa ; Kim, Ji Seok ; Yoon, Yisang ; Santiago, Mayra C. ; Brown, Michael D. ; Park, Joon Young. / Inhibition of Drp1-dependent mitochondrial division impairs myogenic differentiation. In: American Journal of Physiology - Regulatory Integrative and Comparative Physiology. 2013 ; Vol. 305, No. 8. pp. R927-R938.
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