p66(ShcA) and oxidative stress modulate myogenic differentiation and skeletal muscle regeneration after hind limb ischemia.
Oxidative stress plays a pivotal role in ischemic injury, and p66(ShcA)ko mice exhibit both lower oxidative stress and decreased tissue damage following hind limb ischemia. Thus, it was investigated whether tissue regeneration following acute hind limb ischemia was altered in p66(ShcA)ko mice. Upon femoral artery dissection, muscle regeneration started earlier and was completed faster than in wild-type (WT) control. Moreover, faster regeneration was associated with decreased oxidative stress. Unlike ischemia, cardiotoxin injury induced similar skeletal muscle damage in both genotypes. However, p66(ShcA)ko mice regenerated faster, in agreement with the regenerative advantage upon ischemia. Since no difference between p66(ShcA)wt and knock-out (ko) mice was found in blood perfusion recovery after ischemia, satellite cells (SCs), a resident population of myogenic progenitors, were examined. Similar SCs numbers were present in WT and ko mice. However, in vitro cultured p66(ShcA)ko SCs displayed lower oxidative stress levels and higher proliferation rate and differentiated faster than WT. Furthermore, when exposed to sublethal H(2)O(2) doses, p66(ShcA)ko SCs were resistant to H(2)O(2)-induced inhibition of differentiation. Finally, myogenic conversion induced by MyoD overexpression was more efficient in p66(ShcA)ko fibroblasts compared with WT. The present work demonstrates that oxidative stress and p66(ShcA) play a crucial role in the regenerative pathways activated by acute ischemia.
Zaccagnini
Germana
G
Laboratorio di Biologia Vascolare e Terapia Genica, Dipartimento di Chirurgia Vascolare, Centro Cardiologico Monzino--Instituto di Ricovero e Cura a Carattere Scientifico (IRCCS), 20138 Milan, Italy.
Martelli
Fabio
F
Magenta
Alessandra
A
Cencioni
Chiara
C
Fasanaro
Pasquale
P
Nicoletti
Carmine
C
Biglioli
Paolo
P
Pelicci
Pier Giuseppe
PG
Capogrossi
Maurizio C
MC
D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000502:physiology*; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D019085:Fluorescent Antibody Technique, Direct; D006614:Hindlimb / Q000098:blood supply*; D006651:Histocytochemistry; D007511:Ischemia / Q000473:pathology / Q000503:physiopathology*; D008163:Luminescent Measurements; D051379:Mice; D008815:Mice, Inbred Strains; D018345:Mice, Knockout; D008856:Microscopy, Fluorescence; D018482:Muscle, Skeletal / Q000166:cytology / Q000502:physiology*; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism; D012038:Regeneration; D032496:Satellite Cells, Skeletal Muscle / Q000166:cytology / Q000378:metabolism; D055765:Shc Signaling Adaptor Proteins; D013050:Spectrometry, Fluorescence; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D017392:Thiobarbituric Acid Reactive Substances / Q000032:analysis; D013997:Time Factors
D048868:Adaptor Proteins, Signal Transducing; D017382:Reactive Oxygen Species; D055765:Shc Signaling Adaptor Proteins; C528034:Shc1 protein, mouse; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D017392:Thiobarbituric Acid Reactive Substances