NR AWCL
AU Bossers,A.; Harders,F.L.; Smits,M.A.; van Keulen,L.J.M.; van Zijderveld,F.G.
TI Identification of early natural scrapie specific gene expression changes in tonsil and Peyer's patches of sheep using a sheep cDNA microarray
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 GEN-05
PT Konferenz-Poster
AB All currently used routine diagnostics for TSEs are thus far solely based on the detection of pathogically folded accumulated prion protein (PrP). The process of TSE agent-uptake, which most likely takes place in the gut via the ileal Peyer's patches, is still poorly understood, as is the host response to the TSE agent itself. This research focuses on the identification of new biomarkers (other than PrP) that might be useful to extend current TSE diagnostics and/or allowing us to understand TSE pathogenesis better. Therefore, we studied the host response / gene-expression-profiles in the very early phase of a natural scrapie infection in sheep using custom sheep cDNA arrays. We have generated a cDNA microarray of about 32.500 spots per slide containing 7.369 unique features from a large normalised EST library of Tonsil and Peyer's patches of sheep (non-infected and infected of different PrP genotypes). The unique features were determined by sequencing, clustering and assembling over 13.000 sequenced ESTs into 1.850 contigs and 2.594 singletons. Only about 40% for these sequences could be functionally 'annotated' by BLAST to publicly databases thus far. To probe the sheep arrays we collected and mRNA preserved many different tissues that include tonsil, Peyer's patches of ileum and jejunum, brain regions, blood, muscle regions and RLNs. These materials were collected at different time points (0, 3, 8 and 32 weeks) of sheep having different PrP genotypes after natural exposure to scrapie or from scrapie-free sheep. Tissues used for array hybridisation were also checked by IHC for scrapie to be able to link the found gene expression differences to the kinetics of PrPsc deposition. The current status of the gene expression profiling will be presented in relation to PrP genotype, length of incubation, tissue, and PrPsc accumulation.
AD A. Bossers, F.L. Harders, L.J.M. van Keulen: Central Institute for Animal Disease Control (CIDC-Lelystad), The Netherlands; M.A. Smits, F.G. van Zijderveld: Animal Sciences Group, Wageningen UR, The Netherlands. E-mail: alex.bossers@wur.nl
SP englisch
PO Italien