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Different electrophoretic techniques produce conflicting data in the analysis of myocardial samples from dilated cardiomyopathy patients: Protein levels do not necessarily reflect mRNA levels
Author(s) -
Dos Remedios Cristobal G.,
Berry Desirée A.,
Carter Laura K.,
Coumans Joëlle V. F.,
Heinke Monique Y.,
Kiessling Peter C.,
Seeto Reginald K.,
Thorvaldson Tom,
Trahair Toby,
Yeoh Thomas,
Yao Mu,
Gunning Peter W.,
Hardeman Edna,
HumpherySmith Ian,
Naidoo Daya,
Keogh Anne
Publication year - 1996
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150170140
Subject(s) - dystrophin , myosin , dilated cardiomyopathy , blot , biology , microbiology and biotechnology , actin , gel electrophoresis , cardiomyopathy , gene isoform , skeletal muscle , polyacrylamide gel electrophoresis , messenger rna , medicine , heart failure , endocrinology , biochemistry , gene , enzyme
A variety of electrophoretic techniques were used to search for potential causes of human dilated cardiomyopathy (DCM). Northern blots were used to quantify α‐cardiac and α‐skeletal muscle actins, and β‐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.