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Long term anoxia in rainbow trout investigated by 2‐DE and MS/MS
Author(s) -
Wulff Tune,
Jessen Flemming,
Roepstorff Peter,
Hoffmann Else K.
Publication year - 2008
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200700460
Subject(s) - rainbow trout , phosphoglycerate mutase , calpain , cytoskeleton , biology , anoxic waters , trout , microbiology and biotechnology , biochemistry , chemistry , metabolism , cell , fish <actinopterygii> , enzyme , glycolysis , ecology , fishery
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.