Exercise induces myonuclear ubiquitination and apoptosis in dystrophin-deficient muscle of mice.
Apoptosis plays a major role in several diseases, including viral infections, autoimmune diseases, cancer, cardiac infarct, and neurological disorders. To investigate the role of apoptosis in muscular dystrophy, dystrophin-deficient (mdx) mice were subjected to spontaneous exercise and skeletal muscles were analyzed for apoptosis and ubiquitin. The increase of apoptotic myonuclei after exercise was detected by TUNEL, by electron microscopy, and by DNA analyses for high molecular weight and for ladder fragments. Expression of ubiquitin correlated with exercise and with positive myonuclei for apoptosis. Biochemical analysis confirmed a high level of ubiquitination both in sarcoplasmic and myofibrillar proteins. Muscles from sedentary congenit control mice (C57B ) were negative for apoptosis, while after exercise some nuclei were positive. We also revealed that normal myoblasts committed to apoptosis in vitro showed an increased expression of ubiquitin. Western blot for bcl-2, FasL, and BAG1 showed a significant decrease of bcl-2 product only in mdx mice after exercise. Thus, spontaneous exercise results in the increase of ubiquitin expression and in the reduction of bcl-2 tightly related to programmed cell death in mdx mice. These findings confirm that DNA fragmentation, absent in muscles of sedentary normal mice but present in mdx mice at rest, dramatically increases after exercise, shedding new light on exercise-induced muscle damage and on its progression in dystrophinopathies.
Sandri
M
M
Department of Biomedical Sciences, University of Padova, Italy.
Podhorska-Okolow
M
M
Geromel
V
V
Rizzi
C
C
Arslan
P
P
Franceschi
C
C
Carraro
U
U
D000818:Animals; D017209:Apoptosis / Q000235:genetics; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D002478:Cells, Cultured; D053938:DNA Fragmentation; D004268:DNA-Binding Proteins; D016189:Dystrophin / Q000172:deficiency*; D053222:Fas Ligand Protein; D008562:Membrane Glycoproteins / Q000032:analysis; D008565:Membrane Proteins; D051379:Mice; D008810:Mice, Inbred C57BL; D018101:Mice, Inbred mdx; D009124:Muscle Proteins / Q000378:metabolism; D018482:Muscle, Skeletal / Q000378:metabolism / Q000473:pathology*; D009137:Muscular Dystrophy, Animal / Q000378:metabolism / Q000473:pathology*; D005082:Physical Exertion; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000032:analysis; D012420:Running; D014157:Transcription Factors / Q000032:analysis; D014452:Ubiquitins / Q000378:metabolism*
C091732:BCL2-associated athanogene 1 protein; D004268:DNA-Binding Proteins; D016189:Dystrophin; D053222:Fas Ligand Protein; C505658:Fasl protein, mouse; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009124:Muscle Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D014157:Transcription Factors; D014452:Ubiquitins