Comparison of two anoxia models in rainbow trout cells by a 2-DE and MS/MS-based proteome approach.

TY - JOUR

T1 - Comparison of two anoxia models in rainbow trout cells by a 2-DE and MS/MS-based proteome approach.

AU - Wulff, Tune

AU - Hoffmann, Else K

AU - Roepstorff, Peter

AU - Jessen, Flemming

N1 - Keywords: Animals; Cell Hypoxia; Cell Line; Down-Regulation; Electrophoresis, Gel, Two-Dimensional; Fish Proteins; Keratins; Oncorhynchus mykiss; Proteome; Proteomics; Tandem Mass Spectrometry

PY - 2008

Y1 - 2008

N2 - 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.

AB - 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.

U2 - 10.1002/pmic.200700944

DO - 10.1002/pmic.200700944

M3 - Journal article

C2 - 18491317

SN - 1615-9853

VL - 8

SP - 2035

EP - 2044

JO - Proteomics

JF - Proteomics

IS - 10

ER -