Application of fluorescence two-dimensional difference in-gel electrophoresis as a proteomic biomarker discovery tool in muscular dystrophy research.
In this article, we illustrate the application of difference in-gel electrophoresis for the proteomic analysis of dystrophic skeletal muscle. The mdx diaphragm was used as a tissue model of dystrophinopathy. Two-dimensional gel electrophoresis is a widely employed protein separation method in proteomic investigations. Although two-dimensional gels usually underestimate the cellular presence of very high molecular mass proteins, integral membrane proteins and low copy number proteins, this method is extremely powerful in the comprehensive analysis of contractile proteins, metabolic enzymes, structural proteins and molecular chaperones. This gives rise to two-dimensional gel electrophoretic separation as the method of choice for studying contractile tissues in health and disease. For comparative studies, fluorescence difference in-gel electrophoresis has been shown to provide an excellent biomarker discovery tool. Since aged diaphragm fibres from the mdx mouse model of Duchenne muscular dystrophy closely resemble the human pathology, we have carried out a mass spectrometry-based comparison of the naturally aged diaphragm versus the senescent dystrophic diaphragm. The proteomic comparison of wild type versus mdx diaphragm resulted in the identification of 84 altered protein species. Novel molecular insights into dystrophic changes suggest increased cellular stress, impaired calcium buffering, cytostructural alterations and disturbances of mitochondrial metabolism in dystrophin-deficient muscle tissue.
Department of Biology, National University of Ireland, Maynooth, Kildare, Ireland. steven.carberry.is@gmail.com.
Steven
S
Carberry
Department of Physiology II, University of Bonn, Bonn D-53115, Germany. margit.zweyer@ukb.uni-bonn.de.
Margit
M
Zweyer
Department of Physiology II, University of Bonn, Bonn D-53115, Germany. dieter.swandulla@ukb.uni-bonn.de.
Dieter
D
Swandulla
Department of Biology, National University of Ireland, Maynooth, Kildare, Ireland. kay.ohlendieck@nuim.ie.
Kay
K
Ohlendieck