NR AWDS
AU Crecelius,A.; Helmstetter,D.; Mitteregger,G.; Fröhlich,T.; Arnold,G.J.; Kretzschmar,H.A.
TI Comparative proteomic analysis of Prnp 0/0 mouse brain homogenates
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions PA-12
PT Konferenz-Poster
AB The cellular prion protein (PrPc) is a glycoprotein concentrated in neurons of the mammalian brain. Although its function is still elusive, its presence is necessary to generate the conformational pathogenic isoform (PrPsc) in human and animal transmissible spongiform encephalopathies, or prion diseases. The aim of this study is to examine protein changes in Prnp0/0 (Zürich I.) transgenic mouse brain homogenates compared to wild-type homogenates determining the role of PrPc in the brain. Two comparative proteomics techniques, fluorescence 2D difference gel electrophoresis (DIGE) as well as isotope-coded protein labelling (ICPLTM) were applied. For the 2D gel analysis, twenty 2D-DIGE gels with two overlapping pH ranges (pH 4-7 and 6-11) of male, age matched (68 days), wildtype vs. Prnp knockout litter mates (n = 5 per group) were used. No statistical significant differentially expressed proteins (p<0.05, 2-fold change) were obtained by quantitative analysis using DecyderTM 6.5. The results were further confirmed by ICPLTM analysis, in which tryptic mixtures of 13C-labelled Prnp0/0 and 12C labelled wild-type mouse brain samples or vice versa were studied by 2D-nanoLC-MS/MS (Thermo Finnigan LTQ). The data analysis using TurboSequest and ASAPRatio (TransProteomic Pipeline software tools, Insilicos) revealed more then 100 identified ICPL-labelled proteins, but none of them were up- or down regulated by at least a two-fold change. These data agree with studies, in which behavioral alterations between Prnp knockout mice and their controls were only identified, if animals were subjected to abnormal baseline conditions, such as provoked seizures or acute stress.
AD A. Crecelius, D. Helmstetter, G. Mitteregger, H. Kretzschmar: Center of Neuropathology and Prion Research, Laboratory of Molecular Neuropathology, LMU, Munich, Germany; T. Fröhlich, G.J. Arnold: Genecenter,Laboratory for Functional Genome Analysis, LMU, Munich, Germany. E-mail: Anna.Crecelius@med.uni-muenchen.de
SP englisch
PO Italien