NR AXLM
AU Eigenbrod,S.; Eickhoff,M.; Bertsch,U.; Maringer,M.; Mitteregger,G.; Kretzschmar,H.A.
TI A Transgenic Mouse Model for Analyzing the Relevance of Copper Binding Outside the Octarepeat Region in Prion Disease
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Pathology and Pathogenesis P03.87
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Background: Prion protein (PrP) is known to bind copper in the highly conserved octarepeat region. Recent research suggests another strong copper binding site outside the octarepeats at histidine 95 in mice. This latter region is of particular interest because of its essential role in amyloidogenesis.
Objectives: The aim of this study is to gain insight into the pathogenic mechanisms of prion disease, particularly with regard to copper binding outside the octarepeat region in vivo.
Methods: We established a transgenic construct carrying the coding sequence for a mutant PrP protein in which histidine at amino acid 95 is replaced by glycine (H95GPrP) under the regulatory elements of the half-genomic PrP locus. The transgenic construct was used for pronucleus injection into fertilized oocytes and gave rise to five independent transgenic lines. These lines were analyzed in various organs using Western Blot, histology and immunohistochemistry. Infection experiments with scrapie strain RML are in progress.
Results: Western Blot analysis of brain homogenates revealed no differences in band pattern between wildtype- and H95G-mice. Furthermore, no differences were observed after PNGaseF digestion, indicating that the cellular processing of H95G-PrP remains unaffected. Histological and immunohistochemical examinations were also indistinguishable. H95G-mice with PrP expression levels comparable to those of wildtypes were infected with scrapie strain RML and monitored for clinical signs of scrapie and survival. First data suggest a shorter incubation time compared to wildtype mice.
Conclusion: Taken together, those lines are ideal models to study the influence of copper binding outside the octarepeat region of the prion protein in healthy and scrapie infected animals.
AD S. Eigenbrod, M. Eickhoff, U. Bertsch, M. Maringer, G. Mitteregger, H. Kretzschmar, Ludwig-Maximilians-University, Germany
SP englisch
PO Schottland