Pflügers Archiv

, Volume 418, Issue 3, pp 214–219

Effects of ATP antagonists on purinoceptor-operated inward currents in rat phaeochromocytoma cells

  • Ken Nakazawa
  • Kazuhide Inoue
  • Kannosuke Fujimori
  • Akira Takanaka
Excitable Tissues and Central Nervous Physiology

Abstract

The effects of suramin, reactive blue 2 (RB2) and d-tubocurarine (d-TC) were investigated electrophysiologically to elucidate the mechanisms underlying their antagonism of P2 purinoceptor-mediated responses. All three compounds inhibited an adenosine triphosphate (ATP)-activated inward current in rat phaeochromocytoma PC12 cells in a concentration-dependent manner. The order of potency was RB2 > suramin > d-TC. The inhibition induced by suramin or RB2 was reversible, whereas that induced by d-TC was not reversed after a 5-min rinse. The inactivation of the ATP-activated current was accelerated by d-TC but not by suramin or RB2. RB2 administered simultaneously with ATP exerted much weaker inhibition compared to that induced by prior administration, suggesting that RB2 is a slowly acting antagonist. This was not observed for suramin or d-TC. Suramin and RB2 caused a parallel shift in the concentration/response curve for the ATP-activated current. With d-TC the maximal response of ATP was decreased but the concentration producing half-maximal response was unchanged. The voltage dependency of the ATP-activated current showed less inward rectification in the presence of d-TC. Suramin or RB2 did not affect the voltage dependency. These results suggest that suramin and RB2 reversibly block binding of ATP to receptors, whereas d-TC blocks ion permeability through the ATP-activated channel.

Key words

Adenosine triphosphate Purinoceptor Antagonist Non-specific cation channel Phaeochromocytoma cells 

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References

  1. Ascher P, Large WA, Rang HP (1979) Studies on the mechanism of action of acetylcholine antagonists on rat parasympathetic ganglion cells. J Physiol (Lond) 295:139–170Google Scholar
  2. Bean BP (1990) ATP-activated channels in rat and bullfrog sensory neurons: concentration dependence and kinetics. J Neurosci 10:1–10Google Scholar
  3. Bean BP, Friel DD (1990) ATP-activated channels in excitable cells. In: Narahashi T (ed) Ion channels, vol 2. Plenum Press, New York, pp 169–204Google Scholar
  4. Bean BP, Williams CA, Ceelen P (1990) ATP-activated channels in rat and bullfrog sensory neurons: current-voltage relation and single-channel behavior. J Neurosci 10:11–19Google Scholar
  5. Benham CD, Tsien RW (1987) A novel receptor-operated Ca2+-permeable channel activated by ATP in smooth muscle. Nature 328:275–278Google Scholar
  6. Benham CD, Bolton TB, Byrne NG, Large WA (1987) Action of externally applied adenosine triphosphate on single smooth muscle cells from rabbit ear artery. J Physiol (Lond) 387:473–488Google Scholar
  7. Burnstock G, Kennedy C (1985) A dual function for adenosine 5′-triphosphate in the regulation of vascular tone; excitatory cotransmitter with noradrenaline from perivascular nerves and locally released inhibitory intravascular agent. Circ Res 58:319–330Google Scholar
  8. Burnstock G, Warland JJI (1987) P2-purinoceptors of two subtypes in the rabbit mesenteric artery: reactive blue 2 selectively inhibits responses mediated via the P2y but not the P2x-purinoceptor. Br J Pharmacol 90:383–391Google Scholar
  9. Crema A, Frigo GM, Lecchini S, Manzo L, Onori L, Tonini M (1983) Purine receptors in the guinea-pig internal anal sphincter. Br J Pharmacol 78:599–603Google Scholar
  10. Den Hertog A, Nelemans A, Akker JV den (1989) The inhibitory action of suramin on the P2-purinoceptor response in smooth muscle cells of guinea-pig Taenia caeci. Eur J Pharmacol 166:531–534Google Scholar
  11. Dunn PM, Blakeley AGH (1988) Suramin: a reversible P2-purinoceptor antagonist in the mouse vas deferens. Br J Pharmacol 93:243–245Google Scholar
  12. Friel DD (1988) An ATP-sensitive conductance in single smooth muscle cells from the rat vas deferens. J Physiol (Lond) 401:361–380Google Scholar
  13. Gordon L (1986) Extracellular ATP: effects, sources, and fates. Biochem J 233:309–319Google Scholar
  14. Hamill OP, Marty A, Neher E, Sakmann B, Sigworth FJ (1981) Improved patch-clamp techniques for high-resolution current recordings from cells and cell-free membrane patches. Pflügers Arch 391:85–100Google Scholar
  15. Hille B (1984) Endplate channels and other electrically inexcitable channels. In: Ionic channels of excitable membrane. Sinauer, Sunderland, Mass., pp 117–148Google Scholar
  16. Honoré E, Martin C, Mironneau C, Mironneau J (1989) An ATP-sensitive conductance in cultured smooth muscle cells from pregnant rat myometrium. Am J Physiol 257:C297-C305Google Scholar
  17. Hoyle CH, Knight GE, Burnstock G (1990) Suramin antagonizes responses to P2-purinoceptor agonists and purinergic nerve stimulation in the guinea-pig urinary bladder and Taenia coli. Br J Pharmacol 99:617–621Google Scholar
  18. Igusa Y (1988) Adenosine 5′-triphosphate activates acetylcholine receptor channels in cultured Xenopus myotomal muscle cells. J Physiol (Lond) 405:169–185Google Scholar
  19. Inoue K, Kenimer JG (1988) Muscarinic stimulation of calcium influx and norepinephrine release in PC12 cells. J Biol Chem 263:8157–8161Google Scholar
  20. Inoue K, Nakazawa K, Fujimori K, Takanaka A (1989) Extracellular adenosine 5′-triphosphate evoked norepinephrine secretion not relating to voltage-gated Ca channels in pheochromocytoma PC12 cells. Neurosci Lett 106:294–299Google Scholar
  21. Inoue K, Nakazawa K, Ohara-Imaizumi M, Obama T, Fujimori K, Takanaka A (1991a) Selective and competitive antagonism by suramin of ATP-stimulated dopamine-secretion from PC12 phaeochromocytoma cells. Br J Pharmacol (in press)Google Scholar
  22. Inoue K, Nakazawa K, Ohara-Imaizumi M, Obama T, Fujimori K, Takanaka A (1991b) Antagonizm by Reactive Blue 2 but not by Brilliant Blue G of ATP-evoked responses in PC12 phaeochromocytoma cells. Br J Pharmacol (in press)Google Scholar
  23. Jahr CE, Jessell TM (1983) ATP excites a subpopulation of rat dosal horn neurons. Nature 304:730–733Google Scholar
  24. Krishtal OA, Marchenko SM, Pidoplichko VI (1983) Receptor for ATP in the membrane of mammalian sensory neurones. Neurosci Lett 35:41–45Google Scholar
  25. Krishtal OA, Marchenko SM, Obukhov AG (1988a) Receptors for ATP in the membrane of mammalian sensory neurons: the structure-function relationship for ligands. Br J Pharmacol 95:1057–1062Google Scholar
  26. Krishtal OA, Marchenko SM, Obukhov AG, Volkova TM (1988b) Cationic channels activated by extracellular ATP in rat sensory neurons. Neuroscience 27:995–1000Google Scholar
  27. Nakazawa K, Matsuki N (1987) Adenosine triphosphate-activated inward current in isolated smooth muscle cells from rat vas deferens. Pflügers Arch 409:644–646Google Scholar
  28. Nakazawa K, Fujimori K, Takanaka A, Inoue K (1990a) An ATPactivated conductance in pheochromocytoma cells and its suppression by extracellular calcium. J Physiol (Lond) 428:257–272Google Scholar
  29. Nakazawa K, Fujimori K, Takanaka A, Inoue K (1990b) Reversible and selective antagonism by suramin of ATP-activated inward current in PC12 phaeochromocytoma cells. Br J Pharmacol 101:224–226Google Scholar
  30. Rang HP (1982) The action of ganglionic blocking drugs on the synaptic responses of rat submandibular ganglion cells. Br J Pharmacol 75:151–168Google Scholar
  31. Reilly WM, Saville VL, Burnstock G (1987) An assessment of the antagonistic activity of reactive blue 2 at P1 and P2-purinoceptors: supporting evidence for purinergic innervation of the rabbit portal vein. Eur J Pharmacol 140:47–53Google Scholar
  32. Soltoff SP, McMillian MK, Talamo BR (1989) Coomassie brilliant blue G is a more potent antagonist of P2 purinergic responses than reactive blue 2 (cibacron blue 3 GA) in rat parotid acinar cells. Biochem Biophys Res Commun 165:1279–1285Google Scholar
  33. Tallarida RJ, Jacob LS (1979) Kinetis of drug-receptor interaction: interpreting dose-response data. In: Dose-response relation in pharmacology. Springer, Berlin Heidelberg New York, pp 49–84Google Scholar

Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • Ken Nakazawa
    • 1
  • Kazuhide Inoue
    • 1
  • Kannosuke Fujimori
    • 1
  • Akira Takanaka
    • 1
  1. 1.Division of PharmacologyNational Institute of Hygienic SciencesTokyoJapan

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