iTRAQ-coupled 2D LC–MS/MS analysis on differentially expressed proteins in denervated tibialis anterior muscle of Rattus norvegicus
To understand the molecular aspects of denervation-induced atrophy of skeletal muscles, isobaric tags for relative and absolute quantitation (iTRAQ) coupled with two-dimensional liquid chromatography-tandem mass spectrometry were performed to detect a total of 260 proteins that were differentially expressed in the rat tibialis anterior muscle at different times (1, 4, and 8 weeks) after rat sciatic nerve transection. Western blot, gene ontology, and Kyoto Encyclopedia of Genes and Genomes analyses were further conducted for protein validation, functional annotation, and pathway identification, respectively. Among 260 dysregulated proteins, metabolic enzymes represented the largest class of proteins differentially expressed; a down-regulation of β-enolase might be associated with a decreased expression of fast-twitch myosin-4; the 14-3-3 proteins displayed an up-regulation, which might facilitate the inhibition of mTOR signaling; an up-regulation of α-crystallin B chain might be related to the later onset and the slower progress of atrophy; an up-regulation of phosphatidylethanolamine-binding protein-1 perhaps progressively abrogated the cell survival and antiapoptotic properties during muscle atrophy. These results might contribute to the understanding of molecular mechanisms regulating denervation-induced muscle atrophy.
KeywordsSkeletal muscle atrophy Denervation iTRAQ 2D LC–MS/MS
This study was supported by Hi-Tech Research and Development Program of China (863 Program, Grant No. 2006AA02A128), National Natural Science Foundation of China (Grant Nos. 81130080, 81171180, 30870811 and 30670667), a project funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, Research Program of Education Ministry (Grant No. 208044), and Nantong Science and Technology Innovation Program (Grant No. BK2011045). We thank Professor Jie Liu for the assistance in manuscript preparation.
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