Long term anoxia in rainbow trout investigated by 2-DE and MS/MS

Tune Wulff, Flemming Jessen, P. Roepstorff, E.K. Hoffmann

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Twenty-four hours of N-2 induced anoxia induced global perturbations on protein expression in rainbow trout hypodermal fibroblasts cell line. Anoxia was obtained by depleting the medium of O-2 by flushing with N-2, and protein changes were studied by 2-DE coupled with MS providing quantitative measurements of a large number of proteins in one single study. The anoxic insult changed the level of 33 protein spots: 22 of these were up-regulated compared to the control situation and 11 were down-regulated. Using MS/MS sequencing 19 of the 33 protein spots that changed were identified, corresponding to a success rate of more than 50%. The identified proteins included two proteins involved in energy metabolism namely phosphoglycerate mutase and isocitrate dehydrogenase. In addition we observed the up-regulation of a cluster of proteins that contribute to cytoskeleton function. These are calpain, EB1, and Rho GDP dissociation inhibitor (GDI). The up-regulation of Rho GDI was shown to develop in a time dependent manner with no significant increase for up to 8 h of anoxia. In conclusion, this study provides a thorough investigation of the effect of anoxia in a cell line from rainbow trout.
Original languageEnglish
JournalProteomics
Volume8
Issue number5
Pages (from-to)1009-1018
ISSN1615-9853
DOIs
Publication statusPublished - 2008

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