Dystrophin deficient myotubes undergo apoptosis in mouse primary muscle cell culture after DNA damage.
Apoptosis has been demonstrated to occur in differentiated myocardial muscle, neonatal skeletal muscle and skeletal myoblasts in response to injury. In this report, we studied differentiated normal and dystrophin deficient murine skeletal muscle cell cultures that have been injured by a pulse of cis-platinum (2 h). Forty-eight hours after DNA damage, dystrophin positive myotubes appeared almost normal though some myoblasts showed DNA fragmentation. On the other hand, dystrophin deficient myotubes presented progressive degeneration via apoptosis detected either by TUNEL or by nuclear morphology. Degeneration of mdx muscle fibers was confirmed by counting both the number of myotubes observed by contrast phase microscopy and myonuclei viewed by immunoreaction for MyoD. A 6-fold decrease in the number of muscle cells was observed in the dystrophin-deficient cell culture compared to the parental culture (P < 0.001). Direct evidence of degenerating myotubes displaying MyoD- and TUNEL-positive nuclei was obtained. Like myoblasts, differentiated dystrophin deficient myotubes were able to degenerate via apoptosis, showing that mature dystrophin deficient cells are fragile and undergo apoptosis when subjected to a mild injury which would normally be repaired in parental cells.
Sandri
M
M
C.N.R. Unit for Muscle Biology and Physiopathology, Department of Biomedical Sciences, University of Padova, Italy. patgen06@cribi1.bio.unipd.it
Massimino
M L
ML
Cantini
M
M
Giurisato
E
E
Sandri
C
C
Arslan
P
P
Carraro
U
U
D000818:Animals; D000831:Animals, Newborn; D017209:Apoptosis / Q000502:physiology*; D002478:Cells, Cultured; D004249:DNA Damage / Q000502:physiology*; D016189:Dystrophin / Q000032:analysis / Q000172:deficiency*; D020287:In Situ Nick-End Labeling; D051379:Mice; D008810:Mice, Inbred C57BL; D018101:Mice, Inbred mdx; D018485:Muscle Fibers, Skeletal / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism*; D017570:MyoD Protein / Q000032:analysis / Q000378:metabolism
D016189:Dystrophin; D017570:MyoD Protein