NR ARDZ
AU Didier,A.; Dietrich,R.; Boesen,M.; Groschup,M.H.; Pichner,R.; Gareis,M.; Müller,S.; Scherer,S.; Märtlbauer,E.
TI Expression of Cellular Prion Protein in Selected Tissues of Cattle
QU TSE-Forum, 4. Kongress - Nationale TSE-Forschungsplattform, Düsseldorf 28.10.-29.10.2004, Poster GL-08
PT Konferenz-Poster
AB
According to the protein only hypothesis PrPc is the source for conversion to PrPsc during BSE pathogenesis. Therefore knowledge on the quantitative distribution of PrPc is of basic scientific interest and consumers health risk assessment concerns. Interestingly, reports on PrPc distribution are limited and in case of cattle mainly focussed on mRNA expression levels.
To cover this topic on protein level a non-competitive sandwich-enzyme-linked immunosorbent assay has been established. In addition to results presented earlier the study has been extended to a broad spectrum of organs from calves and adult cattle.
After Ribolyser procedure homogenized tissue samples were incubated on EIA plates coated with commercially available monoclonal antibody P4. Samples that had been digested with Proteinase-K served as negative controls. A biotinylated Mab in combination with Extravidin-HRP and TMB as substrate was used as detection system. Absorbance was measured at 450 nm. Optical density given was interpreted in a semiquantitative matter ranging from no to very strong reactivity.
As expected highest and coincident expression of PrPc could be found in brain stem. Comparatively high protein levels were detectable in kidney cortex and lungs of adult cattle. Gut associated lymph nodes as well as lymph nodes of the respiratory tract revealed only weak to intermediated reactivity in EIA. In contrast PrPc content of other organs showed high variability. In particular high variation was observed in liver samples of both calves and adult cattle. Similar results could be found in the third eyelid tissue and in kidney cortex of calves ranging from weak to strong reactivity. In contrast to high and very strong protein expression in thymus of calves respective organ showed predominately intermediate PrPc levels in adults. Here a clear age related difference was obvious.
The project was granted by the Bavarian State Ministry for Environment, Health and Consumer Protection.
AD A. Didier, R. Dietrich, M. Boesen, E. Märtlbauer, Lehrstuhl für Hygiene und Technologie der Milch, LMU, München; M. Groschup, Institut für neue und neuartige Tierseuchenerreger, BFAV, Insel Riems; R. Pichner, M. Gareis, Institut für Mikrobiologie und Toxikologie, BAFF, Kulmbach; S. Müller, S. Scherer, Institut für Mikrobiologie, ZIEL Weihenstephan, Freising
SP englisch
PO Deutschland
OR Tagungsband