Different electrophoretic techniques produce conflicting data in the analysis of myocardial samples from dilated cardiomyopathy patients: protein levels do not necessarily reflect mRNA levels.
A variety of electrophoretic techniques were used to search for potential causes of human dilated cardiomyopathy (DCM). Northern blots were used to quantify alpha-cardiac and alpha-skeletal muscle actins, and beta-myosin heavy chain mRNAs which are the predominant expressed isoform species. We found a wide range of mRNA levels expressed in both DCM and nondiseased (ND) samples of left ventricles. However, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) gels of the same heart samples revealed a stable and constant ratio of actin and myosin. Dystrophin deficiency might account for the DCM symptoms and so dystrophin levels of DCM and ND samples were evaluated using Western blots probed with monoclonal antibodies for the N-, C- and mid-rod portions of this protein. We found that dystrophin levels were constant in all 29 DCM and 5 ND samples suggesting that dystrophin deficiency is probably not a contributing cause. We explored the possibility that terminal failure may be due to an apoptotic-like event in the cardiomyocytes. Zymograms of DCM and ND samples revealed a significant increase in DNase I activity in the DCM group compared to the ND samples. These data raise the possibility that end-stage failure may be associated with apoptosis.
Muscle Research Unit, Department of Anatomy, The University of Sydney, Australia. crisdos@anatomy.su.oz.au
C G
CG
dos Remedios
D A
DA
Berry
L K
LK
Carter
J V
JV
Coumans
M Y
MY
Heinke
P C
PC
Kiessling
R K
RK
Seeto
T
T
Thorvaldson
T
T
Trahair
T
T
Yeoh
M
M
Yao
P W
PW
Gunning
E
E
Hardeman
I
I
Humphery-Smith
D
D
Naidoo
A
A
Keogh
D000199:Actins / Q000032:analysis; D017209:Apoptosis / Q000502:physiology; D015152:Blotting, Northern; D015153:Blotting, Western; D002311:Cardiomyopathy, Dilated / Q000378:metabolism*; D016189:Dystrophin / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D009124:Muscle Proteins / Q000032:analysis*; D018995:Myosin Heavy Chains / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis*; D015203:Reproducibility of Results
D000199:Actins; D016189:Dystrophin; D009124:Muscle Proteins; D012333:RNA, Messenger; D018995:Myosin Heavy Chains