NR ATIA
AU Maringer,M.; Bertsch,U.; Fuhrmann,M.; Piening,N.; Herms,J.W.; Windl,O.; Pilz,C.; Kretzschmar,H.A.
TI Analysis of chimeric EGFP-PrP in transgenic mice
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Cell Biology of PrPc and PrPsc CELL-32
PT Konferenz-Poster
AB
We present transgenic EGFP-PrP mice to study pathogenic mechanisms of prion diseases.
To analyze the localization of prion protein in vivo we established a transgenic construct carrying the coding sequence for an EGFP-PrP fusion protein under the regulatory elements of the half-genomic PrP locus. The fusion protein has been tested for functional EGFP by transfection of RK13 cells and subsequent confocal laser scanning microscopy. Stably transfected cells are used to analyze EGFP-PrP localization before and after infection with the 22L scrapie strain. In addition we performed an in vitro conversion assay using scrapie strain ME7 to investigate, whether EGFP-PrP is convertible to PrPres in vitro. The transgenic construct was used for pronucleus injection into fertilized oocytes and gave rise to five independent transgenic lines, all of which are expressing EGFP-PrP. These lines were analyzed using histological sections from various organs to give a more detailed picture of EGFP-PrP expression in the corresponding line. Different expression patterns of EGFP-PrP were evaluated. Variegation of expression is probably due to integration effects of the transgene. Taken together, those lines are ideal tools to study the localization of EGFP labeled prion protein in vivo in healthy and scrapie-infected animals and in different cell lines derived from transgenic mice. In vivo analysis of EGFP-PrP in various organs of transgenic mice, infection with scrapie strain RML and a functional test regarding disease phenotype rescue in the F35 transgenic mouse line is currently on the way.
AD Marko Maringer, Uwe Bertsch, Martin Fuhrmann, Niklas Piening, Jochen Herms, Otto Windl, Christina Pilz, Hans Kretzschmar, Zentrum für Neuropathologie und Prionforschung LMU München, Germany
SP englisch
PO Deutschland