NR AXWF
AU Sherwood,K.R.; Head,M.W.; Bell,J.W.; Walker,R.; Ironside,J.W.; Fazakerley,J.K.
TI Isolation, Characterisation and Gene Expression Analysis of RNA from Human Autopsy Brain Specimens from UK National CJD Surveillance Unit Tissue Bank
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Pathology and Pathogenesis P03.47
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB One route to increased understanding of the neuropathogenesis of transmissible spongiform encephalopathies is to study how infection alters the pattern of CNS gene expression. We have previously taken this approach in a mouse model of scrapie and identified changes in neuronal and glial cell transcripts during preclinical and clinical disease (Brown et al., Biochem Biophys Res Comm, 334:86-95, 2005). To determine whether these or similar genes also show changes in human prion disease we have isolated RNA from autopsy human brain material from specimens of frontal cortex from vCJD brains held in the UK National CJD Surveillance Unit (Edinburgh) tissue bank and similar material from the UK MRC Sudden Death Brain Bank (Edinburgh). RNA was extracted using Qiagen RNeasy Lipid Tissue Mini kits, followed by additional phenolchloroform extractions to ensure removal of any remaining protein. RNA was analysed for integrity (RNA Integrity Number, RIN) and yield on an Agilent Bioanalyzer 2100 (RNA 6000 nano labchips). A total of 288 RNA samples were generated. Yield and integrity of the RNA did not correlate with age, sex, post-mortem interval or brain pH. RNA of sufficient quality (RIN > 4.5) for quantitative(Q) RT-PCR and gene expression array analysis was recovered from 23% of the samples. Of these, 8 vCJD, 8 other neurological disease and 8 non-neurological disease samples were matched for sex and were closely matched for age (within 10 years). These samples are currently being analysed for gene expression changes using pathway-specific oligonucleotide microarrays (Superarray) including the human Alzheimer's disease Oligo GEArray (which contains 114 pathway specific genes) and Q-PCR.
AD K.R. Sherwood, J.K. Fazakerley, Univeristy of Edinburgh, Center For Infectious Disease, UK; M.W. Head, J.W. Ironside, UK National CJD Surveillance Unit, UK; J.W. Bell, R. Walker, Neuropathology Unit, UK
SP englisch
PO Schottland