Purine receptors and Ca 2+ signalling in the human blood-brain barrier endothelial cell line hCMEC/D3
- verfasst von
- Willem Bintig, Daniela Begandt, Barbara Schlingmann, Linda Gerhard, Maria Pangalos, Lutz Dreyer, Natalija Hohnjec, Pierre Olivier Couraud, Ignacio A. Romero, Babette B. Weksler, Anaclet Ngezahayo
- Abstract
The expression and physiology of purine receptors of the human blood-brain barrier endothelial cells were characterised by application of molecular biological, gene-silencing and Ca 2+-imaging techniques to hCMEC/D3 cells. Reverse transcription polymerase chain reaction showed the expression of the G-protein-coupled receptors P2Y 2-, P2Y 6-, P2Y 11-as well as the ionotropic P2X 4-, P2X 5-and P2X 7-receptors. Fura-2 ratiometry revealed that adenosine triphosphate (ATP) or uridine triphosphate (UTP) mediated a change in the intracellular Ca 2+ concentration ([Ca 2+] i) from 150 to 300 nM in single cells. The change in [Ca 2+] i corresponded to a fourfold to fivefold increase in the fluorescence intensity of Fluo-4, which was used for high-throughput experiments. Pharmacological dissection using different agonists [UTPγS, ATPγS, uridine diphosphate (UDP), adenosine diphosphate (ADP), BzATP, αβ-meATP] and antagonist (MRS2578 or NF340) as well as inhibitors of intracellular mediators (U73122 and 2-APB) showed a PLC-IP 3 cascade-mediated Ca 2+ release, indicating that the nucleotide-induced Ca 2+ signal was mainly related to P2Y 2, 6 and 11 receptors. The gene silencing of the P2Y 2 receptor reduced the ATP-or UTP-induced Ca 2+ signal and suppressed the Ca 2+ signal mediated by P2Y 6 and P2Y 11 more specific agonists like UDP (P2Y 6), BzATP (P2Y 11) and ATPγS (P2Y 11). This report identifies the P2Y 2 receptor subtype as the main purine receptor involved in Ca 2+ signalling of the hCMEC/D3 cells.
- Organisationseinheit(en)
-
Abteilung Pflanzengenomforschung
Institut für Zellbiologie und Biophysik
- Externe Organisation(en)
-
Zentrum für Systemische Neurowissenschaften Hannover (ZSN)
Universite Paris 5
Université de Paris
The Open University
Cornell University
- Typ
- Artikel
- Journal
- Purinergic signalling
- Band
- 8
- Seiten
- 71-80
- Anzahl der Seiten
- 10
- ISSN
- 1573-9538
- Publikationsdatum
- 03.2012
- Publikationsstatus
- Veröffentlicht
- Peer-reviewed
- Ja
- ASJC Scopus Sachgebiete
- Molekularbiologie, Zelluläre und Molekulare Neurowissenschaften, Zellbiologie
- Elektronische Version(en)
-
https://doi.org/10.1007/s11302-011-9262-7 (Zugang:
Eingeschränkt)
