Expression of G-protein subtypes in cultured cerebral endothelial cells

Gabriella Fábián, Csilla Andrea Szabó, Beáta Bozó, John Greenwood, Peter Adamson, M. Deli, Ferenc Joó, I. Krizbai, M. Szücs

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

This paper describes Western-blotting evidence for the presence of various guanine nucleotide binding proteins, G-proteins in cultured rat cerebral endothelial cells (CECs) and two immortalized cerebral endothelial cell lines, RBE4 and GP8. By using specific antibodies raised against known sequences of appropriate G-protein types that were previously characterized, we demonstrated the presence of G5α, G12α, G13α, G(q/11)α, G(o)α and Gβ in cell lysates of primary cultures of CECs, and plasmamembranes of RBE4 and GP8 cells. The appearance of G(o)α proteins in CECs might be of special importance, since they were not detected in peripheral endothelial cells in previous studies. Isoproterenol and bradykinin displayed significant, dose-dependent stimulation of [35S]GTPγS binding above basal values. This assay, reflecting the GDP-GTP exchange reaction on Gα-subunits by receptor agonists, suggested that there were functional, G-protein coupled β-adrenergic and bradykinin receptors in these systems. No significant stimulation of [35S]GTPγS binding was noted with serotonin under our experimental conditions. Since stimulation of [35S]GTPyS binding by isoproterenol and bradykinin was additive, it was concluded that different Gα proteins were activated by these two ligands. In analogy to other systems, activation of G(s) is most likely by isoproterenol, while G(i) and/or Gq/ll proteins might be activated by bradykinin receptors. The possible significance of the receptors and G-proteins detected is being discussed in the functioning of cerebral endothelium, and thus the blood-brain barrier.

Original languageEnglish
Pages (from-to)179-185
Number of pages7
JournalNeurochemistry International
Volume33
Issue number2
DOIs
Publication statusPublished - Aug 1 1998

Fingerprint

GTP-Binding Proteins
Endothelial Cells
Isoproterenol
Bradykinin Receptors
Bradykinin
Gq-G11 GTP-Binding Protein alpha Subunits
Gi-Go GTP-Binding Protein alpha Subunits
Gastrin-Secreting Cells
Guanine Nucleotides
G-Protein-Coupled Receptors
Guanosine Triphosphate
Blood-Brain Barrier
Adrenergic Receptors
Endothelium
Serotonin
Carrier Proteins
Western Blotting
Ligands
Cell Line
Antibodies

Keywords

  • β-adrenergic receptors
  • [S]GTPγS binding
  • Bradykinin
  • Cerebral endothelial cells
  • G-proteins

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Cellular and Molecular Neuroscience

Cite this

Expression of G-protein subtypes in cultured cerebral endothelial cells. / Fábián, Gabriella; Szabó, Csilla Andrea; Bozó, Beáta; Greenwood, John; Adamson, Peter; Deli, M.; Joó, Ferenc; Krizbai, I.; Szücs, M.

In: Neurochemistry International, Vol. 33, No. 2, 01.08.1998, p. 179-185.

Research output: Contribution to journalArticle

Fábián, Gabriella ; Szabó, Csilla Andrea ; Bozó, Beáta ; Greenwood, John ; Adamson, Peter ; Deli, M. ; Joó, Ferenc ; Krizbai, I. ; Szücs, M. / Expression of G-protein subtypes in cultured cerebral endothelial cells. In: Neurochemistry International. 1998 ; Vol. 33, No. 2. pp. 179-185.
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