MilliporeSigma
  • Dynamics of myogenic differentiation using a novel Myogenin knock-in reporter mouse.

Dynamics of myogenic differentiation using a novel Myogenin knock-in reporter mouse.

Skeletal muscle (2021-02-20)
Maria Benavente-Diaz, Glenda Comai, Daniela Di Girolamo, Francina Langa, Shahragim Tajbakhsh
ABSTRACT

Myogenin is a transcription factor that is expressed during terminal myoblast differentiation in embryonic development and adult muscle regeneration. Investigation of this cell state transition has been hampered by the lack of a sensitive reporter to dynamically track cells during differentiation. Here, we report a knock-in mouse line expressing the tdTOMATO fluorescent protein from the endogenous Myogenin locus. Expression of tdTOMATO in MyogntdTom mice recapitulated endogenous Myogenin expression during embryonic muscle formation and adult regeneration and enabled the isolation of the MYOGENIN+ cell population. We also show that tdTOMATO fluorescence allows tracking of differentiating myoblasts in vitro and by intravital imaging in vivo. Lastly, we monitored by live imaging the cell division dynamics of differentiating myoblasts in vitro and showed that a fraction of the MYOGENIN+ population can undergo one round of cell division, albeit at a much lower frequency than MYOGENIN- myoblasts. We expect that this reporter mouse will be a valuable resource for researchers investigating skeletal muscle biology in developmental and adult contexts.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
IGEPAL® CA-630, for molecular biology
Sigma-Aldrich
SIGMAFAST Protease Inhibitor Tablets, For General Use
Sigma-Aldrich
1,4-Bis(2-methylstyryl)benzene, BioReagent, suitable for scintillation, ≥98% (UV)
Sigma-Aldrich
TWEEN® 20, viscous liquid
Sigma-Aldrich
Triton X-100, for molecular biology
Sigma-Aldrich
TWEEN® 20, for molecular biology, viscous liquid