Patterns of expression of muscle-specific markers of differentiation in satellite cell cultures: determination by enzyme-linked immunoculture assay and confocal immunofluorescent assay.
Equine satellite cell clone SE-11 and ovine satellite cell clone I(1)were evaluated for expression of myosin heavy chain, myogenin, desmin, and muscle-specific actin over a 240 h period in culture. An enzyme-linked immunoculture assay (ELICA) was capable of detecting these proteins at all time points evaluated. A linear relationship was demonstrated between the natural logarithm of the absorbance values (corrected for cell number) from the ELICA and percent fusion in both SE-11 and I(1)cultures. The r(2)values for SE-11 cultures were: desmin 0.82, muscle actin 0.81, myogenin 0.78, and myosin 0.70. The r(2)values for I(1)cultures were: desmin 0.77, muscle actin 0.72, myogenin 0.70, and myosin 0.61. Our confocal results support the idea that differences exist between species in the differentiation dynamics of satellite cells. Further, these data suggest that the ELICA may be applied to previously conducted experiments, enabling additional data to be obtained with relation to muscle protein expression.
Stewart
N T
NT
Department of Animal Sciences, Washington State University, Pullman, WA 99164-6310, U.S.A.
Byrne
K M
KM
Ragle
C A
CA
Vierck
J L
JL
Dodson
M V
MV
D000199:Actins / Q000276:immunology / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation; D002459:Cell Fusion; D002999:Clone Cells; D003893:Desmin / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005455:Fluorescent Antibody Technique; D006736:Horses; D007700:Kinetics; D018613:Microscopy, Confocal; D009124:Muscle Proteins / Q000276:immunology / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000378:metabolism* / Q000502:physiology*; D018008:Myogenin / Q000276:immunology / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000276:immunology / Q000378:metabolism; D012756:Sheep; D013234:Stem Cells / Q000166:cytology / Q000378:metabolism
D000199:Actins; D015415:Biomarkers; D003893:Desmin; D009124:Muscle Proteins; D018008:Myogenin; D018995:Myosin Heavy Chains