Identification of protease-activated receptor-4 (PAR-4) in puromycin-purified brain capillary endothelial cells cultured on Matrigel

Szilvia Vajda, Katalin Bartha, Imola Wilhelm, Istvan A. Krizbai, Vera Adam-Vizi

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

An improved method is described for culturing primary rat brain capillary endothelial cells (RBCEC) on glass, covered by Matrigel. The procedure using Matrigel yields spindle-shaped endothelial cells exhibiting close cell-cell appositions seen on electron microscopic sections. These cells permanently express tight junction proteins ZO-1, claudin-5 and the adherent junction protein β-catenin, as revealed by immunofluorescence. Furthermore, glass coverslips covered with Matrigel provide a stable and low-background fluorescent base for microfluorimetric calcium measurements. By this method, hereby we show that the PAR-4 agonist peptide induces transient [Ca2+]i changes with different kinetics compared to that due to activation of the PAR-1 receptor. This indicates that RBCE cells grown on Matrigel express PAR-4 receptors.

Original languageEnglish
Pages (from-to)1234-1239
Number of pages6
JournalNeurochemistry international
Volume52
Issue number6
DOIs
Publication statusPublished - May 1 2008

Keywords

  • Blood-brain barrier
  • Brain endothelial cells
  • Cell culture
  • Microcirculation

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Cell Biology

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