Methods: Using flow cytometry, CD34+/Sca-1-/CD45-/CD31- cells were isolated from extraocular and limb muscle. Pitx2 expression was knocked down in cultured cells using siRNA. Pitx2 transcript and protein levels were assessed with RT-qPCR and western blotting, respectively. The ability of myogenic cells to fuse into multinucleated myotubes was determined with a fusion index assay.
Results: Pitx2 transcript and protein levels are higher in cells from EOM compared to limb muscle. Loss of Pitx2 expression reduced the ability of myogenic cells from EOM and limb muscle to form multinucleated myotubes.
Conclusions: Pitx2 enhances the fusion of myogenic cells in EOM. The higher expression of Pitx2 in EOM cells may contribute to their sparing in muscular dystrophy.
Keywords: Cell culture, Gene expression, Molecular biology, Muscle and Proteins
See more of: Craniofacial Biology