Characterization of P2X3, P2Y1 and P2Y4 receptors in cultured HEK293-hP2X3 cells and their inhibition by ethanol and trichloroethanol

Wolfgang Fischer, Kerstin Wirkner, Marco Weber, Christoph Eberts, Laszlo Köles, Robert Reinhardt, Heike Franke, Clemens Allgaier, Clemens Gillen, Peter Illes

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Abstract

Membrane currents and changes in the intracellular Ca2+ concentration ([Ca2+]i) were measured in HEK293 cells transfected with the human P2X3 receptor (HEK293-hP2X3). RT-PCR and immunocytochemistry indicated the additional presence of endogenous P2Y1 and to some extent P2Y4 receptors. P2 receptor agonists induced inward currents in HEK293-hP2X3 cells with the rank order of potency α,β-meATP ≈ ATP > ADP-β-S > UTP. A comparable rise in [Ca2+]i was observed after the slow superfusion of ATP, ADP-β-S and UTP; α,β-meATP was ineffective. These data, in conjunction with results obtained by using the P2 receptor antagonists TNP-ATP, PPADS and MRS2179 indicate that the current response to α,β-meATP is due to P2X3 receptor activation, while the ATP-induced rise in [Ca2+]i is evoked by P2Y1 and P2Y4 receptor activation. TCE depressed the α,β-meATP current in a manner compatible with a non-competitive antagonism. The ATP-induced increase of [Ca2+]i was much less sensitive to the inhibitory effect of TCE than the current response to α,β-meATP. The present study indicates that in HEK293-hP2X3 cells, TCE, but not ethanol, potently inhibits ligand-gated P2X3 receptors and, in addition, moderately interferes with G protein-coupled P2Y1 and P2Y4 receptors. Such an effect may be relevant for the interruption of pain transmission in dorsal root ganglion neurons following ingestion of chloral hydrate or trichloroethylene.

Original languageEnglish
Pages (from-to)779-790
Number of pages12
JournalJournal of neurochemistry
Volume85
Issue number3
DOIs
Publication statusPublished - May 2003

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Keywords

  • Ethanol
  • HEK293 cells
  • P2X receptor
  • P2Y receptor
  • Trichloroethanol

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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