Balázs, B.; Dankó, T.; Kovács, G.; Köles, L.; Hediger, M. A.; Zsembery, Á. (2013). Investigation of the inhibitory effects of the benzodiazepine derivative, 5-BDBD on P2X4 purinergic receptors by two complementary methods. Cellular physiology and biochemistry, 32(1), pp. 11-24. Karger 10.1159/000350119
|
Text
000350119.pdf - Published Version Available under License Creative Commons: Attribution-Noncommercial (CC-BY-NC). Download (3MB) | Preview |
BACKGROUND/AIMS
ATP-gated P2X4 purinergic receptors (P2X4Rs) are cation channels with important roles in diverse cell types. To date, lack of specific inhibitors has hampered investigations on P2X4Rs. 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 P2X4Rs.
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 Ca(2+) 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.
Item Type: |
Journal Article (Original Article) |
|---|---|
Division/Institute: |
04 Faculty of Medicine > Pre-clinic Human Medicine > Institute of Biochemistry and Molecular Medicine |
UniBE Contributor: |
Kovacs, Gergely, Hediger, Matthias |
Subjects: |
500 Science > 570 Life sciences; biology 600 Technology > 610 Medicine & health |
ISSN: |
1015-8987 |
Publisher: |
Karger |
Language: |
English |
Submitter: |
Patrizia Catucci |
Date Deposited: |
01 May 2014 09:30 |
Last Modified: |
25 May 2023 15:13 |
Publisher DOI: |
10.1159/000350119 |
PubMed ID: |
23867750 |
BORIS DOI: |
10.48350/43303 |
URI: |
https://boris.unibe.ch/id/eprint/43303 |
Actions (login required)
 |
Edit item |