Lack of run-down of smooth muscle P2X receptor currents recorded with the amphotericin permeabilized patch technique, physiological and pharmacological characterization of the properties of mesenteric artery P2X receptor ion channels
Article first published online: 29 JAN 2009
2000 British Pharmacological Society
British Journal of Pharmacology
Volume 131, Issue 8, pages 1659–1666, December 2000
How to Cite
Lewis, C. J. and Evans, R. J. (2000), Lack of run-down of smooth muscle P2X receptor currents recorded with the amphotericin permeabilized patch technique, physiological and pharmacological characterization of the properties of mesenteric artery P2X receptor ion channels. British Journal of Pharmacology, 131: 1659–1666. doi: 10.1038/sj.bjp.0703744
- Issue published online: 29 JAN 2009
- Article first published online: 29 JAN 2009
- (Received August 3, 2000, Revised September 26, 2000, Accepted October 2, 2000)
- P2X receptors;
- P2X1 receptors;
- patch clamp;
Immunoreactivity for P2X1, P2X4 and P2X5 receptor subtypes was detected in the smooth muscle cell layer of second and third order rat mesenteric arteries immunoreactivity, for P2X2, P2X3, P2X6 and P2X7 receptors was below the level of detection in the smooth muscle layer.
P2X receptor-mediated currents were recorded in patch clamp studies on acutely dissociated mesenteric artery smooth muscle cells. Purinergic agonists evoked transient inward currents that decayed rapidly in the continued presence of agonist (τ∼200 ms). Standard whole cell responses to repeated applications of agonist at 5 min intervals ran down. Run-down was unaffected by changes in extracellular calcium concentration, intracellular calcium buffering or the inclusion of ATP and GTP in the pipette solution.
Run-down was overcome and reproducible responses to purinergic agonists were recorded using the amphotericin permeabilized patch recording configuration.
The rank order of potency at the P2X receptor was ATP=2 methylthio ATP>α,β-methylene ATP>CTP=l-β,γ-methylene ATP. Only ATP and 2meSATP were full agonists. The P2 receptor antagonists suramin and PPADS inhibited P2X receptor-mediated currents with IC50s of 4 μM and 70 nM respectively.
These results provide further characterization of artery P2X receptors and demonstrate that the properties are dominated by a P2X1-like receptor phenotype. No evidence could be found for a phenotype corresponding to homomeric P2X4 or P2X5 receptors or to heteromeric P2X1/5 receptors and the functional role of these receptors in arteries remains unclear.
British Journal of Pharmacology (2000) 131, 1659–1666; doi:10.1038/sj.bjp.0703744