Life-Long Transgene Expression in Skeletal Muscle Without Transduction of Satellite Cells Following Embryonic Myogenic Progenitor Transduction by Lentivirus Administered in Utero.
Embryologic events in mammalian myogenesis remain to be fully defined. Recent evidence supports the presence of a common progenitor arising in the dermomyotome that gives rise to both embryologic and adult muscle and postnatal myogenic stem cells (satellite cells). In this study, we utilize the technique of early intra-amniotic gene transfer to target nascent muscle progenitors as they traverse the primitive streak before formation of the dermomyotome. This technique robustly transduced both epaxial and hypaxial muscle groups. Marker gene expression is observed in up to 100% muscle fibers in the lower extremities and is sustained for the lifetime of the mouse. We next analyzed transduced muscle for satellite cell transduction using highly sensitive methodology. Surprisingly, despite high levels of sustained transgene expression in muscle fibers, satellite cells lacked the marker transgene. Our data suggest that dermatomyotome is a heterogeneous structure and that not all myogenic progenitors of dermatomyotome give rise to satellite cells.
1 Department of Pediatric Surgery, Yale School of Medicine , New Haven, Connecticut.
David H
DH
Stitelman
2 Department of Pediatric General, Thoracic and Fetal Surgery, The Children's Hospital of Philadelphia , Philadelphia, Pennsylvania.
Tim R
TR
Brazelton
2 Department of Pediatric General, Thoracic and Fetal Surgery, The Children's Hospital of Philadelphia , Philadelphia, Pennsylvania.
Masayuki
M
Endo
2 Department of Pediatric General, Thoracic and Fetal Surgery, The Children's Hospital of Philadelphia , Philadelphia, Pennsylvania.
Archana
A
Bora
2 Department of Pediatric General, Thoracic and Fetal Surgery, The Children's Hospital of Philadelphia , Philadelphia, Pennsylvania.
Jeremy
J
Traas
2 Department of Pediatric General, Thoracic and Fetal Surgery, The Children's Hospital of Philadelphia , Philadelphia, Pennsylvania.
Philip W
PW
Zoltick
2 Department of Pediatric General, Thoracic and Fetal Surgery, The Children's Hospital of Philadelphia , Philadelphia, Pennsylvania.
Alan W
AW
Flake
D000818:Animals; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism*; D051787:Myogenic Regulatory Factor 5 / Q000235:genetics* / Q000378:metabolism; D032496:Satellite Cells, Skeletal Muscle / Q000378:metabolism*; D019076:Transgenes
C497418:Myf5 protein, mouse; D051787:Myogenic Regulatory Factor 5