NR AXWE
AU Serrano,C.; Harders,F.L.; Lyahyai,J.; Bolea,R.; Badiola,J.J.; Zaragoza,P.; Martin-Burriel,I.; Bossers,A.
TI Analysis of Gene Expression Profiles in Cns of Naturally Scrapie Infected Sheep Using a Sheep cDNA Microarray
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Pathology and Pathogenesis P03.153
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
The underlying mechanisms of scrapie pathogenesis and many neurodegenerative diseases are still poorly understood. The identification of genes with differential expression in CNS of infected animals might provide clues to clarify the molecular mechanisms that lead to neuronal loss, being useful for future therapies and to identify molecular biomarkers that might be the basis for new diagnostic tests. We present here an initial study on the transcriptional differences in cerebellum obtained from naturally infected Scrapie sheep using cDNA microarray hybridizations. We have used the sheep cDNA microarray generated at CIDC-Lelystad (communication of Bossers et al., Prion2006). Total RNA of cerebellum was isolated from 5 control sheep and 9 infected sheep. According to IHC characteristics on cerebellum (CER), spleen (SP) and mesentery lymph node (MN) of infected sheep their RNAs were grouped into 4 pools
(1: +CER, +SP, +MN; 2: -CER, -SP, -MN; 3: +CER, -/+SP, -MN; 4: -CER, +SP, +MN). The remaining (5) pool was formed with the 5 controls. The five RNA pools were hybridized against a universal reference RNA, after cDNA synthesis and fluorescent labeling. We compared "in silico" gene expression of the 4 positive groups against the control group. One common clone was identified to be up-regulated within the 4 diseased groups and two clones identified as down-regulated. In the two groups with PrPsc deposit on cerebellum (1 and 3) we found 2 up-expressed clones and 4 downexpressed clones in common. The other two groups (2 and 4) shared 6 clones with a significant expression increase and 3 clones with decreased expression. The differentiated clones were compared with the GenBank database and some of them showed similarity with known human and bovine genes. Small sequences of genes GMPS, RPL32 and ATP6AP2 align with down-expressed clones. By contrast, the genes GNB2L1, HSPA8, RPS3 and FN1 have similarity with up-expressed sequences. The expression of 11 common clones has been analyzed by Real Time PCR in order to confirm these previous results. Moreover, the expression of these genes has been analyzed in 5 controls and 9 scrapie animals; GMPS and two un-known sequences showed significant differences between both groups.
AD C. Serrano, J. Lyahyai, P. Zaragoza, I. Martín-Burriel, Facultad de Veterinaria, Anatomía, Embriología y Genética Animal, Spain; F.L. Harders, A. Bossers, Central Institute for Animal Disease Control (CIDC-Lelystad, Wageningen UR), Bacteriology and TSEs, Netherlands; R. Bolea, J.J. Badiola, Facultad de Veterinaria, Centro de Investigaciones Priónicas, Spain
SP englisch
PO Schottland