Skip to main content
British Journal of Pharmacology logoLink to British Journal of Pharmacology
. 1990 Jul;100(3):619–625. doi: 10.1111/j.1476-5381.1990.tb15856.x

The properties of the ATP-induced depolarization and current in single cells isolated from the guinea-pig urinary bladder.

R Inoue 1, A F Brading 1
PMCID: PMC1917811  PMID: 1697199

Abstract

1. The actions of exogenously applied ATP were investigated with the whole-cell patch clamp method in single cells isolated from guinea-pig urinary bladder with a modified concentration jump technique. 2. Rapid application of ATP (threshold ca. 100 nM) depolarized the cell membrane with superimposition of action potentials which was followed by transient hyperpolarization. In the presence of D600, the amplitude of the ATP-induced depolarization was a function of the ATP concentration (EC50: 0.5-1 microM). 3. ATP activated a dose-dependent inward current with a short latency (18 ms with 10 microM ATP; measured as the time between the start of application and 10% of the peak). The relationship of the peak current versus ATP concentration was well fitted by a Michaelis-Menten equation with a Hill coefficient (n) of 1.7 and a dissociation constant (Kd) of 2.3 microM. The current desensitized rapidly and the time course of desensitization was a function of the ATP concentration and could be fitted by two exponentials. 4. The reversal potential of the ATP-activated current was near 0 mV. Replacement of extracellular Na by other monovalent or divalent cations indicated that the current flows through nonselective cation channels. 5. alpha,beta-Methylene ATP also produced a dose-dependent inward current but was less potent than ATP (n: 1.6, Kd: 10.4 microM). alpha,beta-Methylene ATP blocked the response to ATP by desensitization of the receptor. 6. alpha,beta-Methylene ATP was 50-100 times more potent than ATP at eliciting a contractile response of strips of detrusor smooth muscle. 7. The relevance of the above results to the possible role of ATP as the fast excitatory transmitter is discussed.

Full text

PDF
620

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Akaike N., Maruyama T., Tokutomi N. Kinetic properties of the pentobarbitone-gated chloride current in frog sensory neurones. J Physiol. 1987 Dec;394:85–98. doi: 10.1113/jphysiol.1987.sp016861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Benham C. D. ATP-activated channels gate calcium entry in single smooth muscle cells dissociated from rabbit ear artery. J Physiol. 1989 Dec;419:689–701. doi: 10.1113/jphysiol.1989.sp017893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benham C. D., Bolton T. B., Byrne N. G., Large W. A. Action of externally applied adenosine triphosphate on single smooth muscle cells dispersed from rabbit ear artery. J Physiol. 1987 Jun;387:473–488. doi: 10.1113/jphysiol.1987.sp016585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Benham C. D., Tsien R. W. A novel receptor-operated Ca2+-permeable channel activated by ATP in smooth muscle. Nature. 1987 Jul 16;328(6127):275–278. doi: 10.1038/328275a0. [DOI] [PubMed] [Google Scholar]
  5. Brading A. F., Mostwin J. L. Electrical and mechanical responses of guinea-pig bladder muscle to nerve stimulation. Br J Pharmacol. 1989 Dec;98(4):1083–1090. doi: 10.1111/j.1476-5381.1989.tb12651.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brock J. A., Cunnane T. C. Electrical activity at the sympathetic neuroeffector junction in the guinea-pig vas deferens. J Physiol. 1988 May;399:607–632. doi: 10.1113/jphysiol.1988.sp017099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burnstock G., Kennedy C. Is there a basis for distinguishing two types of P2-purinoceptor? Gen Pharmacol. 1985;16(5):433–440. doi: 10.1016/0306-3623(85)90001-1. [DOI] [PubMed] [Google Scholar]
  8. Burnstock G. Purinergic nerves. Pharmacol Rev. 1972 Sep;24(3):509–581. [PubMed] [Google Scholar]
  9. Byrne N. G., Large W. A. Comparison of the biphasic excitatory junction potential with membrane responses to adenosine triphosphate and noradrenaline in the rat anococcygeus muscle. Br J Pharmacol. 1984 Nov;83(3):751–758. doi: 10.1111/j.1476-5381.1984.tb16229.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Creed K. E., Ishikawa S., Ito Y. Electrical and mechanical activity recorded from rabbit urinary bladder in response to nerve stimulation. J Physiol. 1983 May;338:149–164. doi: 10.1113/jphysiol.1983.sp014666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Franke C., Hatt H., Dudel J. Liquid filament switch for ultra-fast exchanges of solutions at excised patches of synaptic membrane of crayfish muscle. Neurosci Lett. 1987 Jun 15;77(2):199–204. doi: 10.1016/0304-3940(87)90586-6. [DOI] [PubMed] [Google Scholar]
  12. Friel D. D. An ATP-sensitive conductance in single smooth muscle cells from the rat vas deferens. J Physiol. 1988 Jul;401:361–380. doi: 10.1113/jphysiol.1988.sp017167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Friel D. D., Bean B. P. Two ATP-activated conductances in bullfrog atrial cells. J Gen Physiol. 1988 Jan;91(1):1–27. doi: 10.1085/jgp.91.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fujii K. Evidence for adenosine triphosphate as an excitatory transmitter in guinea-pig, rabbit and pig urinary bladder. J Physiol. 1988 Oct;404:39–52. doi: 10.1113/jphysiol.1988.sp017277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hamill O. P., Marty A., Neher E., Sakmann B., Sigworth F. J. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflugers Arch. 1981 Aug;391(2):85–100. doi: 10.1007/BF00656997. [DOI] [PubMed] [Google Scholar]
  16. Honoré E., Martin C., Mironneau C., Mironneau J. An ATP-sensitive conductance in cultured smooth muscle cells from pregnant rat myometrium. Am J Physiol. 1989 Aug;257(2 Pt 1):C297–C305. doi: 10.1152/ajpcell.1989.257.2.C297. [DOI] [PubMed] [Google Scholar]
  17. Horn R., Marty A. Muscarinic activation of ionic currents measured by a new whole-cell recording method. J Gen Physiol. 1988 Aug;92(2):145–159. doi: 10.1085/jgp.92.2.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hoyle C. H., Burnstock G. Atropine-resistant excitatory junction potentials in rabbit bladder are blocked by alpha,beta-methylene ATP. Eur J Pharmacol. 1985 Aug 15;114(2):239–240. doi: 10.1016/0014-2999(85)90635-1. [DOI] [PubMed] [Google Scholar]
  19. Inoue R., Kitamura K., Kuriyama H. Acetylcholine activates single sodium channels in smooth muscle cells. Pflugers Arch. 1987 Sep;410(1-2):69–74. doi: 10.1007/BF00581898. [DOI] [PubMed] [Google Scholar]
  20. Ishikawa S. Actions of ATP and alpha, beta-methylene ATP on neuromuscular transmission and smooth muscle membrane of the rabbit and guinea-pig mesenteric arteries. Br J Pharmacol. 1985 Dec;86(4):777–787. doi: 10.1111/j.1476-5381.1985.tb11099.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Karim F., Majid D. S., Summerill R. A. Sympathetic nerves in the mediation of renal response to localized stimulation of atrial receptors in anaesthetized dogs. J Physiol. 1989 Oct;417:63–78. doi: 10.1113/jphysiol.1989.sp017790. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kasakov L., Burnstock G. The use of the slowly degradable analog, alpha, beta-methylene ATP, to produce desensitisation of the P2-purinoceptor: effect on non-adrenergic, non-cholinergic responses of the guinea-pig urinary bladder. Eur J Pharmacol. 1982 Dec 24;86(2):291–294. doi: 10.1016/0014-2999(82)90330-2. [DOI] [PubMed] [Google Scholar]
  23. Klöckner U., Isenberg G. Action potentials and net membrane currents of isolated smooth muscle cells (urinary bladder of the guinea-pig). Pflugers Arch. 1985 Dec;405(4):329–339. doi: 10.1007/BF00595685. [DOI] [PubMed] [Google Scholar]
  24. Klöckner U., Isenberg G. Calcium currents of cesium loaded isolated smooth muscle cells (urinary bladder of the guinea pig). Pflugers Arch. 1985 Dec;405(4):340–348. doi: 10.1007/BF00595686. [DOI] [PubMed] [Google Scholar]
  25. Krishtal O. A., Pidoplichko V. I. A receptor for protons in the nerve cell membrane. Neuroscience. 1980;5(12):2325–2327. doi: 10.1016/0306-4522(80)90149-9. [DOI] [PubMed] [Google Scholar]
  26. Langley J. N., Anderson H. K. The Innervation of the Pelvic and adjoining Viscera: Part II. The Bladder. Part III. The External Generative Organs. Part IV. The Internal Generative Organs. Part V. Position of the Nerve Cells on the Course of the Efferent Nerve Fibres. J Physiol. 1895 Dec 30;19(1-2):71–139. doi: 10.1113/jphysiol.1895.sp000587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Miledi R., Parker I. Latencies of membrane currents evoked in Xenopus oocytes by receptor activation, inositol trisphosphate and calcium. J Physiol. 1989 Aug;415:189–210. doi: 10.1113/jphysiol.1989.sp017718. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nakazawa K., Matsuki N. Adenosine triphosphate-activated inward current in isolated smooth muscle cells from rat vas deferens. Pflugers Arch. 1987 Aug;409(6):644–646. doi: 10.1007/BF00584668. [DOI] [PubMed] [Google Scholar]
  29. Northway M. G., Burks T. F. Stimulation of cholinergic nerves in dog intestine by adenine nucleotides. Eur J Pharmacol. 1980 Jul 11;65(1):11–19. doi: 10.1016/0014-2999(80)90203-4. [DOI] [PubMed] [Google Scholar]
  30. Strange P. G. The structure and mechanism of neurotransmitter receptors. Implications for the structure and function of the central nervous system. Biochem J. 1988 Jan 15;249(2):309–318. doi: 10.1042/bj2490309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tsien R. W. Calcium channels in excitable cell membranes. Annu Rev Physiol. 1983;45:341–358. doi: 10.1146/annurev.ph.45.030183.002013. [DOI] [PubMed] [Google Scholar]

Articles from British Journal of Pharmacology are provided here courtesy of The British Pharmacological Society

RESOURCES