Divide and conquer: rat liver tissue proteomics based on the analysis of purified constituents.
Comparative proteome data of normal and diseased tissue samples are difficult to interpret. Proteins detected in tissues are derived from different cell types and blood constituents. Pathologic or toxicant-induced aberrations may affect the proteome profile of tissues in several ways since different cell types may respond in very different and highly specific manners. The aim of this study was to analyze the proteome profiles of purified rat liver primary cells and of blood plasma in comparison to liver whole tissue. Moreover, we investigated alterations of these profiles induced by the liver toxicant N-nitrosomorpholine (NNM) used as a model compound. Whole liver samples, pure hepatocytes and Kupffer cells as well as blood plasma were obtained from saline- or NNM-treated rats. Proteins were separated by 2-D PAGE and their amounts were estimated by fluorography. Selected proteins were identified by MS analysis of tryptic digests. Among them we identified proteins exclusively expressed in the analysed constituents. Several of these proteins were assigned in the proteome profile of whole-tissue homogenates. Furthermore, we identified several proteins that were modified, up-regulated or down-regulated due to NNM treatment in total liver homogenates. Some of these protein alterations were specifically detected in primary cells isolated from NNM-treated rats. Thus, we demonstrated the successful assignment of NNM-induced proteome alterations in rat liver to the cell type of origin. The currently applied approach may help to better understand pathologic processes at a whole-tissue level.
Teufelhofer
Olga
O
Internal Medicine Clinic I, Institute of Cancer Research, Medical University of Vienna, Vienna, Austria.
Parzefall
Wolfram
W
Elbling
Leonilla
L
Kainzbauer
Eveline
E
Grasl-Kraupp
Bettina
B
Zielinski
Christoph
C
Schulte-Hermann
Rolf
R
Gerner
Christopher
C
D000818:Animals; D002469:Cell Separation; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D022781:Hepatocytes / Q000737:chemistry; D007728:Kupffer Cells / Q000737:chemistry; D008099:Liver / Q000737:chemistry* / Q000187:drug effects; D008297:Male; D009602:Nitrosamines / Q000633:toxicity; D040901:Proteomics / Q000379:methods*; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results
D009602:Nitrosamines; C002741:N-nitrosomorpholine