An examination of MS candidate genes identified as differentially regulated in multiple sclerosis plaque tissue, using absolute and comparative real-time Q-PCR analysis.
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We have developed methods in real time detection of quantitative-polymerase chain reaction (Q-PCR) to analyse the relative levels of gene expression in post mortem brain tissue. We have then examined differences in gene activity between normal white matter (NWM) and plaque tissue from multiple sclerosis (MS) patients, displaying different pathologies (either acute or chronic active). The result from each MS plaque was then compared with an age and sex matched NWM control, obtained from the same anatomical location. The results of comparative and absolute Q-PCR analysis compared favourably (P<0.05, Pearsons), with significant differences in gene expression between chronic active and acute pathologies identified for four of the five genes examined. A 20-60-fold increase in osteopontin (SPP1) and inositol 1-4-5 phosphate 3 kinase B (ITPKB) levels was identified in acute plaques compared with NWM controls. This contrasted with a 5-fold (or less) increase in expression in chronic active plaques (P<0.05, unpaired t-Test). Both the signal transducer and activator of transcription (STAT)1 and the protein inhibitor of activated STAT1 (PIAS) displayed (in contrast) a 5-10 fold decrease in expression in acute tissues, with no significant difference in chronic active tissues, compared with NWM. Additionally, no association was identified with changes in Calpain (CAPNS1) levels and MS pathology. In summary, Q-PCR analysis has allowed the identification of clinically significant associations between gene expression and pathology from small amounts of CNS tissue; which may be used as the basis for further clinical investigations.
Brain Research Protocols
Copyright 2005 Elsevier : Reproduced in accordance with the copyright policy of the publisher : This journal is available online - use hypertext links.