Isobaric Tagging-Based Quantification for Proteomic Analysis: A Comparative Study of Spared and Affected Muscles from mdx Mice at the Early Phase of Dystrophy

PLoS One. 2013 Jun 18;8(6):e65831. doi: 10.1371/journal.pone.0065831. Print 2013.

Abstract

Duchenne muscular dystrophy (DMD) is the most common childhood myopathy, characterized by muscle loss and cardiorespiratory failure. While the genetic basis of DMD is well established, secondary mechanisms associated with dystrophic pathophysiology are not fully clarified yet. In order to obtain new insights into the molecular mechanisms of muscle dystrophy during earlier stages of the disease, we performed a comparative proteomic profile of the spared extraocular muscles (EOM) vs. affected diaphragm from the mdx mice, using a label based shotgun proteomic approach. Out of the 857 identified proteins, 42 to 62 proteins had differential abundance of peptide ions. The calcium-handling proteins sarcalumenin and calsequestrin-1 were increased in control EOM compared with control DIA, reinforcing the view that constitutional properties of EOM are important for their protection against myonecrosis. The finding that galectin-1 (muscle regeneration), annexin A1 (anti-inflammatory) and HSP 47 (fibrosis) were increased in dystrophic diaphragm provides novel insights into the mechanisms through which mdx affected muscles are able to counteract dystrophy, during the early stage of the disease. Overall, the shotgun technique proved to be suitable to perform quantitative comparisons between distinct dystrophic muscles and allowed the suggestion of new potential biomarkers and drug targets for dystrophinopaties.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Female
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Inbred mdx
  • Muscle, Skeletal / metabolism
  • Muscle, Skeletal / pathology*
  • Muscular Dystrophy, Duchenne / metabolism
  • Muscular Dystrophy, Duchenne / pathology*
  • Proteomics*

Grant support

This work was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, grants 11/51697-6; 08/58491-1) and the Swedish Research Council. MJM is recipient of fellowships from Conselho Nacional de Pesquisas (301306/2010-9). CYM was recipient of a fellowship from Fundação de Amparo à Pesquisa do Estado de São Paulo (grant 08/54775-5) and from CAPES (grant 2014-10-6). MS data were obtained from the Proteomics Core Facility, Gothenburg University, on equipment obtained by a grant from Knut and Alice Wallenberg (KAW grant 2007.0118). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.