A robust Pax7EGFP mouse that enables the visualization of dynamic behaviors of muscle stem cells

• DOI: 10.1186/s13395-018-0169-7
• Publication Date: 2018-08-21T14:22:21Z
• Authors: Elisia D. Tichy; David K. Sidibe; Christopher D. Greer; Nicholas M. Oyster; Panteleimon Rompolas; Nadia A. Rosenthal; Helen M. Blau; Foteini Mourkioti
• Keywords: Cell Sorting; Immunofluorescence

Pax7 is a transcription factor involved in the specification and maintenance of muscle stem cells (MuSCs). Upon injury, MuSCs leave their quiescent state, downregulate differentiate, contributing to skeletal regeneration. In majority regeneration studies, are isolated by fluorescence-activated sorting (FACS), based on cell surface markers. It known that heterogeneous population only small percentage true able self-renew. A strong reporter line would be valuable study vivo behavior Pax7-expressing cells.We generated characterized properties new transgenic Pax7EGFP mouse. Utilizing traditional immunofluorescence assays, we analyzed whole embryos sections fluorescence microscopy, addition muscles 2-photon detect specificity EGFP expression. Skeletal from mice were also evaluated steady state under injury conditions. Finally, MuSCs-derived control sorted FACS myogenic activity was comparatively examined.Our studies provide with robust expression, detectable both flow cytometry microscopy. Pax7EGFP-derived have identical wild-type MuSCs, vitro vivo, excluding any positional effect due transgene insertion. Furthermore, demonstrated high label temporal manner recapitulates reported expression pattern. Interestingly, analysis showed marks satellite position adult fixed live tissues.This mouse could an invaluable tool for variety questions related MuSC biology, including but not limited heterogeneity, polarity, aging, regeneration, motility, either itself or combination harboring additional genetic alterations.