Comparison of two anoxia models in rainbow trout cells by a 2-DE and MS/MS-based proteome approach.
In the literature, a variety of ways have been used to obtain anoxia, and most often results are compared between studies without taking into consideration how anoxia has been obtained. Here, we provide a comprehensive study of two types of anoxia, using a proteomics approach to compare changes in protein expression. The two investigated situations were 30 min of chemical anoxia (10 mM NaN(3)) followed by reoxygenation overnight (CR) and 2 h of N(2)-induced anoxia (achieved by flushing with N(2)) followed by reoxygenation overnight (NR), after which samples were resolved by 2-DE. Forty-five protein spots changed their abundance in response to CR and 35 protein spots changed their abundance in response to NR, but only six proteins changed their abundance in response to both stimuli. By the means of MS/MS, 40 protein spots were identified including proteins involved in processes like cell protection and protein synthesis. It was also revealed that the level of a number of keratins was down-regulated. This study therefore provides a valuable comparison of two different anoxia models and shows that great care should be taken when comparing the effects of anoxia in studies that have used different types and durations of anoxia.
Wulff
Tune
T
Department of Biochemistry, Institute of Molecular Biology, University of Copenhagen, Copenhagen, Denmark. tuw@difres.dk
Hoffmann
Else K
EK
Roepstorff
Peter
P
Jessen
Flemming
F
D000818:Animals; D015687:Cell Hypoxia; D002460:Cell Line; D015536:Down-Regulation; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D029941:Fish Proteins / Q000032:analysis*; D007633:Keratins / Q000032:analysis / Q000378:metabolism; D017686:Oncorhynchus mykiss; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D029941:Fish Proteins; D020543:Proteome; D007633:Keratins