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 prote...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2013-12-01
|
| Series: | Biology |
| Subjects: | |
| Online Access: | http://www.mdpi.com/2079-7737/2/4/1438 |
| _version_ | 1797725644100993024 |
|---|---|
| author | Steven Carberry Margit Zweyer Dieter Swandulla Kay Ohlendieck |
| author_facet | Steven Carberry Margit Zweyer Dieter Swandulla Kay Ohlendieck |
| author_sort | Steven Carberry |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-12T10:34:14Z |
| format | Article |
| id | doaj.art-ffea8ed381b84818900c3d960f7e399b |
| institution | Directory Open Access Journal |
| issn | 2079-7737 |
| language | English |
| last_indexed | 2024-03-12T10:34:14Z |
| publishDate | 2013-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biology |
| spelling | doaj.art-ffea8ed381b84818900c3d960f7e399b2023-09-02T09:01:36ZengMDPI AGBiology2079-77372013-12-01241438146410.3390/biology2041438biology2041438Application of Fluorescence Two-Dimensional Difference In-Gel Electrophoresis as a Proteomic Biomarker Discovery Tool in Muscular Dystrophy ResearchSteven Carberry0Margit Zweyer1Dieter Swandulla2Kay Ohlendieck3Department of Biology, National University of Ireland, Maynooth, Kildare, IrelandDepartment of Physiology II, University of Bonn, Bonn D-53115, GermanyDepartment of Physiology II, University of Bonn, Bonn D-53115, GermanyDepartment of Biology, National University of Ireland, Maynooth, Kildare, IrelandIn 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.http://www.mdpi.com/2079-7737/2/4/1438cvHspdiaphragmDIGEDuchenne muscular dystrophydystrophinopathygel electrophoresisHspB7mdxmouse modelparvalbumin |
| spellingShingle | Steven Carberry Margit Zweyer Dieter Swandulla Kay Ohlendieck Application of Fluorescence Two-Dimensional Difference In-Gel Electrophoresis as a Proteomic Biomarker Discovery Tool in Muscular Dystrophy Research Biology cvHsp diaphragm DIGE Duchenne muscular dystrophy dystrophinopathy gel electrophoresis HspB7 mdx mouse model parvalbumin |
| title | Application of Fluorescence Two-Dimensional Difference In-Gel Electrophoresis as a Proteomic Biomarker Discovery Tool in Muscular Dystrophy Research |
| title_full | Application of Fluorescence Two-Dimensional Difference In-Gel Electrophoresis as a Proteomic Biomarker Discovery Tool in Muscular Dystrophy Research |
| title_fullStr | Application of Fluorescence Two-Dimensional Difference In-Gel Electrophoresis as a Proteomic Biomarker Discovery Tool in Muscular Dystrophy Research |
| title_full_unstemmed | Application of Fluorescence Two-Dimensional Difference In-Gel Electrophoresis as a Proteomic Biomarker Discovery Tool in Muscular Dystrophy Research |
| title_short | Application of Fluorescence Two-Dimensional Difference In-Gel Electrophoresis as a Proteomic Biomarker Discovery Tool in Muscular Dystrophy Research |
| title_sort | application of fluorescence two dimensional difference in gel electrophoresis as a proteomic biomarker discovery tool in muscular dystrophy research |
| topic | cvHsp diaphragm DIGE Duchenne muscular dystrophy dystrophinopathy gel electrophoresis HspB7 mdx mouse model parvalbumin |
| url | http://www.mdpi.com/2079-7737/2/4/1438 |
| work_keys_str_mv | AT stevencarberry applicationoffluorescencetwodimensionaldifferenceingelelectrophoresisasaproteomicbiomarkerdiscoverytoolinmusculardystrophyresearch AT margitzweyer applicationoffluorescencetwodimensionaldifferenceingelelectrophoresisasaproteomicbiomarkerdiscoverytoolinmusculardystrophyresearch AT dieterswandulla applicationoffluorescencetwodimensionaldifferenceingelelectrophoresisasaproteomicbiomarkerdiscoverytoolinmusculardystrophyresearch AT kayohlendieck applicationoffluorescencetwodimensionaldifferenceingelelectrophoresisasaproteomicbiomarkerdiscoverytoolinmusculardystrophyresearch |