NR AWBJ
AU Banner,S.J.; Brown,D.; Thomson,V.; Clarke,A.R.; Manson,J.; Gill,A.C.
TI Analysis of murine CNS proteomes of PrP Knockout and WT mice
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 PA-04
PT Konferenz-Poster
AB The prion protein (PrP) is fundamental to TSE disease biology and its conversion from the normal, cellular form (PrPc) to a detergent insoluble and protease resistant isoform (PrPsc) appears to be a pre-requisite for disease progression. Aggregates that accumulate in TSE disease are strongly immunopositive for PrPsc leading to the suggestion that PrPsc aggregation may be responsible for neurodegeneration via a 'gain of function' mechanism. However, in some TSE cases, extensive pathology can exist despite the absence of detectable levels of PrPsc. The role that PrPsc has in neuropathogenesis is therefore unclear and an alternate hypothesis suggests that the loss of PrPc function during disease progression could be responsible for neurodegeneration. We hypothesis that PrPc may function as a neuroprotective molecule and believe that mutations in the PrnP gene could initiate pathological disease due to impaired functioning of PrPc. The normal biological role of PrPc is still unclear and hence transgenic mice devoid of PrPc (PrP0/0) were developed in order to address this point. Knockout PrP mice do show subtle defects in synaptic transmission, mitochondrial function and circadian rhythm and an initial, collaborative, microarray based pilot study of wildtype (WT) versus PrP0/0 mice uncovered several intriguing differences between them. Our ongoing work intends to build on this preliminary data and aims to define more specifically the temporal molecular changes in PrP0/0 mice and establish whether mutant PrP, with a reduced neuroprotective function, can cause similar changes. Here we present preliminary steps to undertake proteomic analyses of WT and PrP0/0 mice taken at different time points using traditional 2D-PAGE techniques. Proteins solubilised from mouse hemisphere and cerebellum have been subjected to isoelectric focusing and then separated by SDS PAGE. Proteins have been visualised by silver staining and gel images digitised. Comparative analyses are currently underway and we aim to present our initial data. Using this approach we aim to confirm the microarray data by assessing if the changes seen in mRNA levels are replicated in protein expression changes. Furthermore, we intend to investigate whether similar changes can be seen in transgenic mice expressing mutant PrP molecules. Ultimately we hope to confirm and define a role for PrPc in neuroprotection during ageing.
AD S.J. Banner, A.C. Gill: Institute for Animal Health, Compton Laboratory, Compton, Berkshire, RG207NN, UK; D. Brown, V. Thomson, J. Manson: Institute for Animal Health, Neuropathogenesis Unit, Ogston Building, West Mains Rd, Edinburgh EH93JF, UK; A.R. Clarke: Cardiff University, Biomedical Sciences Building, Cardiff, CF103US, UK. E-mail: steve.banner@bbsrc.ac.uk
SP englisch
PO Italien