Abstract
We have compared the induced expression of E-selectin in primary cultures of rat brain microvascular endothelial cells (EC), pericytes and in non-CNS microvascular endothelium stimulated with the cytokines, IL-1β (20 ng/ml), and tumor necrosis factor (TNF)-α (75 ng/ml). Expression was studied at both the protein and mRNA levels. Fluorescence in-situ hybridization (FISH) was used to examine de novo synthesis of E-selectin mRNA. Laser cytometric analysis was used as a novel approach to the quantitaion of FISH. In-situ hybridization was performed using two PCR-generated probes. The first probe (517 bp) spanned the lectin and epidermal growth factor (EGF)-like domain. The second probe (562 bp) spanned the CR3, 4, and 6 domains. E-selectin-specific mRNA was localized to the perinuclear regions of the EC. Both cytokines, IL-1β and TNF-α significantly increased E-selectin gene expression in CNS EC but not pericytes. IL-1β induced higher E-selectin mRNA levels than TNF-α. The maximum number of mRNA-positive cells was observed after stimulation for 4-6 h. Surface protein expression was sustained for up to 48 h following addition of cytokine. This was in contrast to the transient expression in non-CNS EC indicating that pure primary CNS EC display slightly different kinetics of E-selectin expression than non-CNS EC.
Original language | English (US) |
---|---|
Pages (from-to) | 51-62 |
Number of pages | 12 |
Journal | Journal of the Neurological Sciences |
Volume | 195 |
Issue number | 1 |
DOIs | |
State | Published - Mar 15 2002 |
Keywords
- Central nervous system
- Cytokines
- E-selectin
- Endothelial cells
- Gene regulation
- Interleukin-1
- Microvessels
- Tumor necrosis factor (TNF)
ASJC Scopus subject areas
- Clinical Neurology
- Neurology