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Rotenone inhibits primary murine myotube formation via Raf-1 and ROCK2

Sander Grefte1, Jori A L Wagenaars1, Renate Jansen1

  • 1Department of Biochemistry, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, Nijmegen, The Netherlands.

Biochimica Et Biophysica Acta
|April 2, 2015
PubMed

Insights

Rotenone specifically inhibits myotube formation by affecting the Raf-1/ROCK2 pathway, independent of its impact on mitochondrial complex I. This finding offers new insights into rotenone

Area of Science:

  • Mitochondrial biology and cellular respiration
  • Muscle development and differentiation

Background:

  • Rotenone (ROT) is a common inhibitor of mitochondrial complex I (CI) in the oxidative phosphorylation (OXPHOS) system.
  • ROT can exhibit off-target effects, particularly at higher concentrations, necessitating investigation into its precise mechanisms of action.

Purpose of the Study:

  • To investigate the specific effects of rotenone on in vitro primary murine myotube formation.
  • To elucidate the molecular pathways involved in rotenone-induced inhibition of myogenesis.

Main Methods:

  • Treatment of primary murine myotubes with rotenone (ROT) and piericidin A (PA).
  • Assessment of myotube formation and myoblast oxygen consumption.
  • Gene knockdown experiments targeting ROCK1, ROCK2, and RhoA, along with inhibition of Raf-1 activity.

Main Results:

  • ROT (10-100nM) specifically inhibited myotube formation, unlike piericidin A (PA).
  • Both ROT and PA blocked oxygen consumption at 100nM.
  • ROCK2 knockdown, and to a lesser extent ROCK1, prevented ROT-induced inhibition, which was reversed by inhibiting Raf-1 activity.
  • RhoA knockdown did not prevent ROT-induced inhibition of myotube formation.

Conclusions:

  • Rotenone inhibits primary myotube formation through a mechanism independent of its effect on Complex I-driven mitochondrial ATP production.
  • The Raf-1/ROCK2 pathway is critically involved in rotenone's inhibitory effects on myogenesis.
  • These findings highlight specific off-target effects of rotenone in muscle cell differentiation.
Keywords:
Fusion indexGW5074Piericidin ARho-GTPaseRotenoneU0126

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