Acta Neuropathologica

, Volume 119, Issue 5, pp 601–615

Gene expression analysis of the microvascular compartment in multiple sclerosis using laser microdissected blood vessels

  • Paula Cunnea
  • Jill McMahon
  • Enda O’Connell
  • Kaveh Mashayekhi
  • Una Fitzgerald
  • Stephen McQuaid
Original Paper

Abstract

The blood brain barrier (BBB) is formed by capillary endothelial cells with inter-endothelial cell tight junctions and other cells such as pericytes and astrocytes present. Previous studies have shown a role for tight junction abnormalities in BBB leakage in multiple sclerosis (MS) brain. This marks a key stage in the development of inflammatory demyelination in MS. The aim of this study was to identify aberrantly expressed genes involved in BBB changes in MS lesions. A focused endothelial cell biology microarray, capable of detecting changes in expression of 113 endothelial cell-specific genes, was employed to analyse endothelial cell mRNA extracted from post-mortem control white matter, MS normal appearing white matter (NAWM), chronic active or inactive lesions by laser capture microdissection. Microarray analysis found 52 genes out of 113 analysed, predominantly in the activation functional group, to be differentially expressed in lesions compared to control or NAWM (p < 0.01). The majority of the differentially expressed genes were validated by quantitative real time PCR. In addition, the protein expression profiles of ICAM2, MMP2, and VEGFR1 were examined by immunofluorescent staining of selected tissue blocks. ICAM-2 was expressed at a higher level in chronic inactive lesions than control or NAWM, corresponding with the increased mRNA measured by microarray and real time PCR. The data shown, presenting a number of differentially expressed genes in the microvascular compartment of MS lesions, may shed light on the molecular mechanisms that are involved in the breakdown of the BBB. This moves us a step closer to the identification of potential therapeutic targets for repair of the compromised BBB.

Keywords

Microvascular compartment Multiple sclerosis Laser capture microdissection Endothelial microarray 

Supplementary material

401_2009_618_MOESM1_ESM.doc (42 kb)
Supplementary material 1 (DOC 42 kb)

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Paula Cunnea
    • 1
  • Jill McMahon
    • 2
  • Enda O’Connell
    • 2
  • Kaveh Mashayekhi
    • 4
  • Una Fitzgerald
    • 2
  • Stephen McQuaid
    • 1
    • 3
  1. 1.Centre for Cancer Research and Cell Biology, School of Biomedical SciencesQueen’s University BelfastBelfastNorthern Ireland, UK
  2. 2.National Centre for Biomedical Engineering and SciencesNational University of Ireland GalwayGalwayRepublic of Ireland
  3. 3.Tissue PathologyBelfast Health and Social Care TrustBelfastNorthern Ireland, UK
  4. 4.Regenerative Medicine InstituteNational University of Ireland GalwayGalwayRepublic of Ireland

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