Cytokine regulation of E-selectin in rat CNS microvascular endothelial cells: Differential response of CNS and non-CNS vessels

Jarmila Wagnerova, Larisa Cervenakova, Roumen Balabanov, Ian Zitron, Paula Dore-Duffy*

*Corresponding author for this work

Research output: Contribution to journalArticle

11 Scopus citations

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 languageEnglish (US)
Pages (from-to)51-62
Number of pages12
JournalJournal of the Neurological Sciences
Volume195
Issue number1
DOIs
StatePublished - 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

  • Neurology
  • Clinical Neurology

Fingerprint Dive into the research topics of 'Cytokine regulation of E-selectin in rat CNS microvascular endothelial cells: Differential response of CNS and non-CNS vessels'. Together they form a unique fingerprint.

  • Cite this