Maria Benavente-Diaz1,2,3, Glenda Comai1,2, Daniela Di Girolamo1,2, Francina Langa4, Shahragim Tajbakhsh5,6. 1. Stem Cells & Development Unit, Institut Pasteur, 25 rue du Dr. Roux, 75015, Paris, France. 2. UMR CNRS 3738, Institut Pasteur, Paris, France. 3. Sorbonne Universités, Complexité du Vivant, F-75005, Paris, France. 4. Mouse Genetics Engineering Center, Institut Pasteur, Paris, France. 5. Stem Cells & Development Unit, Institut Pasteur, 25 rue du Dr. Roux, 75015, Paris, France. shahragim.tajbakhsh@pasteur.fr. 6. UMR CNRS 3738, Institut Pasteur, Paris, France. shahragim.tajbakhsh@pasteur.fr.
Abstract
BACKGROUND: 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. RESULTS: 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. CONCLUSIONS: We expect that this reporter mouse will be a valuable resource for researchers investigating skeletal muscle biology in developmental and adult contexts.
BACKGROUND: 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. RESULTS: 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. CONCLUSIONS: We expect that this reporter mouse will be a valuable resource for researchers investigating skeletal muscle biology in developmental and adult contexts.
Authors: C I Rodríguez; F Buchholz; J Galloway; R Sequerra; J Kasper; R Ayala; A F Stewart; S M Dymecki Journal: Nat Genet Date: 2000-06 Impact factor: 38.330
Authors: Ramkumar Sambasivan; Roseline Yao; Adrien Kissenpfennig; Laetitia Van Wittenberghe; Andràs Paldi; Barbara Gayraud-Morel; Hind Guenou; Bernard Malissen; Shahragim Tajbakhsh; Anne Galy Journal: Development Date: 2011-09 Impact factor: 6.868