Investigation of the inhibitory effects of the benzodiazepine derivative, 5-BDBD on P2X4 purinergic receptors by two complementary methods

Bernadett Balázs, Tamás Dankó, Gergely Kovács, L. Köles, Matthias A. Hediger, A. Zsembery

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Background/Aims: ATP-gated P2X4 purinergic receptors (P2X 4Rs) are cation channels with important roles in diverse cell types. To date, lack of specific inhibitors has hampered investigations on P2X 4Rs. Recently, the benzodiazepine derivative, 5-BDBD has been proposed to selectively inhibit P2X4Rs. However, limited evidences are currently available on its inhibitory properties. Thus, we aimed to characterize the inhibitory effects of 5-BDBD on recombinant human P2X 4Rs. Methods: We investigated ATP-induced intracellular Ca 2+ signals and whole cell ion currents in HEK 293 cells that were either transiently or stably transfected with hP2X4Rs. Results: Our data show that ATP (<1 μM) stimulates P2X4R-mediated Ca 2+ influx while endogenously expressed P2Y receptors are not activated to any significant extent. Both 5-BDBD and TNP-ATP inhibit ATP-induced Ca2+ signals and inward ion currents in a concentration-dependent manner. Application of two different concentrations of 5-BDBD causes a rightward shift in ATP dose-response curve. Since the magnitude of maximal stimulation does not change, these data suggest that 5-BDBD may competitively inhibit the P2X4Rs. Conclusions: Our results demonstrate that application of submicromolar ATP concentrations allows reliable assessment of recombinant P2XR functions in HEK 293 cells. Furthermore, 5-BDBD and TNP-ATP have similar inhibitory potencies on the P2X4Rs although their mechanisms of actions are different.

Original languageEnglish
Pages (from-to)11-24
Number of pages14
JournalCellular Physiology and Biochemistry
Volume32
Issue number1
DOIs
Publication statusPublished - Aug 2013

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Purinergic P2X4 Receptors
Benzodiazepines
Adenosine Triphosphate
HEK293 Cells
Ions
Cations

Keywords

  • 5-BDBD
  • ATP
  • Calcium infux
  • Electrophysiology
  • P2X receptors

ASJC Scopus subject areas

  • Physiology

Cite this

Investigation of the inhibitory effects of the benzodiazepine derivative, 5-BDBD on P2X4 purinergic receptors by two complementary methods. / Balázs, Bernadett; Dankó, Tamás; Kovács, Gergely; Köles, L.; Hediger, Matthias A.; Zsembery, A.

In: Cellular Physiology and Biochemistry, Vol. 32, No. 1, 08.2013, p. 11-24.

Research output: Contribution to journalArticle

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