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Vajda S, Bartha K, Wilhelm I, Krizbai IA, Adam-Vizi V.
Identification of protease-activated receptor-4 (PAR-4) in puromycin-purified
brain capillary endothelial cells cultured on Matrigel.
Neurochem Int. 2008 May;52(6):1234-9
Department of Medical Biochemistry, Semmelweis University, Neurobiochemical
Group, Hungarian Academy of Sciences, Szentagothai Knowledge Center, Budapest,
Hungary.
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 beta-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.
PMID: 18294734 |
