Skip to main content
NCBI home page
The Journal of Physiology logoLink to The Journal of Physiology
. 1988 Sep;403:621–640. doi: 10.1113/jphysiol.1988.sp017268

Inhibition of the calcium channel by intracellular protons in single ventricular myocytes of the guinea-pig.

M Kaibara 1, M Kameyama 1
PMCID: PMC1190732  PMID: 2855346

Abstract

1. The inhibitory effects of intracellular protons (Hi+) on the L-type Ca2+ channel activity were investigated in single ventricular myocytes of guinea-pigs by using the patch-clamp method in the open-cell-attached patch configuration, where 'run down' of the channel was partially prevented. 2. Hi+ reduced the unitary Ba2+ current of the Ca2+ channel by 10-20% without changing the maximum slope conductance. 3. Hi+ did not alter the number of channels in patches containing one or two channels. 4. Hi+ markedly reduced the mean current normalized by the unitary current, which gave the open-state probability multiplied by the number of channels in the patch. The dose-response curve between Hi+ and the open-state probability indicated half-maximum inhibition at pHi 6.6 and an apparent Hill coefficient of 1. 5. Hi+ shifted both the steady-state activation and inactivation curves in a negative direction by 10-15 mV, and the effects were reversible. 6. Hi+ did not affect the fast open-closed kinetics represented by the C-C-O scheme, apart from increasing the slow time constant of the closed time. 7. Hi+ increased the percentage of blank sweeps and reduced that of non-blank sweeps resulting in a decreased probability of channel opening. 8. Photo-oxidation with Rose Bengal abolished the reducing effect of Hi+ on the open-state probability (Po) in two out of ten experiments, suggesting the possible involvement of histidine residues in the Hi+ effect. 9. The above results indicate that Hi+ inhibits the Ba2+ current mainly by affecting the slow gating mechanism of the channel.

Full text

PDF
621

Selected References

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

  1. Baumgold J., Matsumoto G., Tasaki I. Biochemical studies of nerve excitability: the use of protein modifying reagents for characterizing sites involved in nerve excitation. J Neurochem. 1978 Jan;30(1):91–100. doi: 10.1111/j.1471-4159.1978.tb07039.x. [DOI] [PubMed] [Google Scholar]
  2. Bean B. P., Nowycky M. C., Tsien R. W. Beta-adrenergic modulation of calcium channels in frog ventricular heart cells. 1984 Jan 26-Feb 1Nature. 307(5949):371–375. doi: 10.1038/307371a0. [DOI] [PubMed] [Google Scholar]
  3. Brown A. M., Camerer H., Kunze D. L., Lux H. D. Similarity of unitary Ca2+ currents in three different species. Nature. 1982 Sep 9;299(5879):156–158. doi: 10.1038/299156a0. [DOI] [PubMed] [Google Scholar]
  4. Brown A. M., Lux H. D., Wilson D. L. Activation and inactivation of single calcium channels in snail neurons. J Gen Physiol. 1984 May;83(5):751–769. doi: 10.1085/jgp.83.5.751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brum G., Osterrieder W., Trautwein W. Beta-adrenergic increase in the calcium conductance of cardiac myocytes studied with the patch clamp. Pflugers Arch. 1984 Jun;401(2):111–118. doi: 10.1007/BF00583870. [DOI] [PubMed] [Google Scholar]
  6. Cachelin A. B., de Peyer J. E., Kokubun S., Reuter H. Ca2+ channel modulation by 8-bromocyclic AMP in cultured heart cells. Nature. 1983 Aug 4;304(5925):462–464. doi: 10.1038/304462a0. [DOI] [PubMed] [Google Scholar]
  7. Cavalié A., Ochi R., Pelzer D., Trautwein W. Elementary currents through Ca2+ channels in guinea pig myocytes. Pflugers Arch. 1983 Sep;398(4):284–297. doi: 10.1007/BF00657238. [DOI] [PubMed] [Google Scholar]
  8. Cavalié A., Pelzer D., Trautwein W. Fast and slow gating behaviour of single calcium channels in cardiac cells. Relation to activation and inactivation of calcium-channel current. Pflugers Arch. 1986 Mar;406(3):241–258. doi: 10.1007/BF00640910. [DOI] [PubMed] [Google Scholar]
  9. Chandler W. K., Hodgkin A. L., Meves H. The effect of changing the internal solution on sodium inactivation and related phenomena in giant axons. J Physiol. 1965 Oct;180(4):821–836. doi: 10.1113/jphysiol.1965.sp007733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chesnais J. M., Coraboeuf E., Sauviat M. P., Vassas J. M. Sensitivity to H, Li and Mg ions of the slow inward sodium current in frog atrial fibres. J Mol Cell Cardiol. 1975 Sep;7(9):627–642. doi: 10.1016/0022-2828(75)90140-6. [DOI] [PubMed] [Google Scholar]
  11. Clark H. R., Strickholm A. Evidence for a conformational change in nerve membrane with depolarization. Nature. 1971 Dec 24;234(5330):470–471. doi: 10.1038/234470a0. [DOI] [PubMed] [Google Scholar]
  12. Fenwick E. M., Marty A., Neher E. Sodium and calcium channels in bovine chromaffin cells. J Physiol. 1982 Oct;331:599–635. doi: 10.1113/jphysiol.1982.sp014394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hagiwara S., Ohmori H. Studies of single calcium channel currents in rat clonal pituitary cells. J Physiol. 1983 Mar;336:649–661. doi: 10.1113/jphysiol.1983.sp014603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hagiwara S., Ozawa S., Sand O. Voltage clamp analysis of two inward current mechanisms in the egg cell membrane of a starfish. J Gen Physiol. 1975 May;65(5):617–644. doi: 10.1085/jgp.65.5.617. [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. Hescheler J., Kameyama M., Trautwein W. On the mechanism of muscarinic inhibition of the cardiac Ca current. Pflugers Arch. 1986 Aug;407(2):182–189. doi: 10.1007/BF00580674. [DOI] [PubMed] [Google Scholar]
  17. Hess P., Lansman J. B., Tsien R. W. Different modes of Ca channel gating behaviour favoured by dihydropyridine Ca agonists and antagonists. Nature. 1984 Oct 11;311(5986):538–544. doi: 10.1038/311538a0. [DOI] [PubMed] [Google Scholar]
  18. Hille B. Charges and potentials at the nerve surface. Divalent ions and pH. J Gen Physiol. 1968 Feb;51(2):221–236. doi: 10.1085/jgp.51.2.221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Horie M., Irisawa H., Noma A. Voltage-dependent magnesium block of adenosine-triphosphate-sensitive potassium channel in guinea-pig ventricular cells. J Physiol. 1987 Jun;387:251–272. doi: 10.1113/jphysiol.1987.sp016572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Iijima T., Ciani S., Hagiwara S. Effects of the external pH on Ca channels: experimental studies and theoretical considerations using a two-site, two-ion model. Proc Natl Acad Sci U S A. 1986 Feb;83(3):654–658. doi: 10.1073/pnas.83.3.654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Irisawa H., Sato R. Intra- and extracellular actions of proton on the calcium current of isolated guinea pig ventricular cells. Circ Res. 1986 Sep;59(3):348–355. doi: 10.1161/01.res.59.3.348. [DOI] [PubMed] [Google Scholar]
  22. Kakei M., Noma A., Shibasaki T. Properties of adenosine-triphosphate-regulated potassium channels in guinea-pig ventricular cells. J Physiol. 1985 Jun;363:441–462. doi: 10.1113/jphysiol.1985.sp015721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kameyama M., Hescheler J., Hofmann F., Trautwein W. Modulation of Ca current during the phosphorylation cycle in the guinea pig heart. Pflugers Arch. 1986 Aug;407(2):123–128. doi: 10.1007/BF00580662. [DOI] [PubMed] [Google Scholar]
  24. Kameyama M., Hescheler J., Mieskes G., Trautwein W. The protein-specific phosphatase 1 antagonizes the beta-adrenergic increase of the cardiac Ca current. Pflugers Arch. 1986 Oct;407(4):461–463. doi: 10.1007/BF00652635. [DOI] [PubMed] [Google Scholar]
  25. Kameyama M., Kakei M., Sato R., Shibasaki T., Matsuda H., Irisawa H. Intracellular Na+ activates a K+ channel in mammalian cardiac cells. Nature. 1984 May 24;309(5966):354–356. doi: 10.1038/309354a0. [DOI] [PubMed] [Google Scholar]
  26. Kohlhardt M., Haap K., Figulla H. R. Influence of low extracellular pH upon the Ca inward current and isometric contractile force in mammalian ventricular myocardium. Pflugers Arch. 1976 Oct 15;366(1):31–38. doi: 10.1007/BF02486557. [DOI] [PubMed] [Google Scholar]
  27. Kokubun S., Prod'hom B., Becker C., Porzig H., Reuter H. Studies on Ca channels in intact cardiac cells: voltage-dependent effects and cooperative interactions of dihydropyridine enantiomers. Mol Pharmacol. 1986 Dec;30(6):571–584. [PubMed] [Google Scholar]
  28. Kokubun S., Reuter H. Dihydropyridine derivatives prolong the open state of Ca channels in cultured cardiac cells. Proc Natl Acad Sci U S A. 1984 Aug;81(15):4824–4827. doi: 10.1073/pnas.81.15.4824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kurachi Y. The effects of intracellular protons on the electrical activity of single ventricular cells. Pflugers Arch. 1982 Sep;394(3):264–270. doi: 10.1007/BF00589102. [DOI] [PubMed] [Google Scholar]
  30. Lansman J. B., Hess P., Tsien R. W. Blockade of current through single calcium channels by Cd2+, Mg2+, and Ca2+. Voltage and concentration dependence of calcium entry into the pore. J Gen Physiol. 1986 Sep;88(3):321–347. doi: 10.1085/jgp.88.3.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Nilius B., Hess P., Lansman J. B., Tsien R. W. A novel type of cardiac calcium channel in ventricular cells. Nature. 1985 Aug 1;316(6027):443–446. doi: 10.1038/316443a0. [DOI] [PubMed] [Google Scholar]
  32. Noma A., Shibasaki T. Membrane current through adenosine-triphosphate-regulated potassium channels in guinea-pig ventricular cells. J Physiol. 1985 Jun;363:463–480. doi: 10.1113/jphysiol.1985.sp015722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Noma A., Tsuboi N. Dependence of junctional conductance on proton, calcium and magnesium ions in cardiac paired cells of guinea-pig. J Physiol. 1987 Jan;382:193–211. doi: 10.1113/jphysiol.1987.sp016363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Nowycky M. C., Fox A. P., Tsien R. W. Three types of neuronal calcium channel with different calcium agonist sensitivity. Nature. 1985 Aug 1;316(6027):440–443. doi: 10.1038/316440a0. [DOI] [PubMed] [Google Scholar]
  35. Ochi R., Hino N., Okuyama H. Beta-adrenergic modulation of the slow gating process of cardiac calcium channels. Jpn Heart J. 1986 Nov;27 (Suppl 1):51–55. [PubMed] [Google Scholar]
  36. Reimann E. M., Walsh D. A., Krebs E. G. Purification and properties of rabbit skeletal muscle adenosine 3',5'-monophosphate-dependent protein kinases. J Biol Chem. 1971 Apr 10;246(7):1986–1995. [PubMed] [Google Scholar]
  37. Sanguinetti M. C., Kass R. S. Voltage-dependent block of calcium channel current in the calf cardiac Purkinje fiber by dihydropyridine calcium channel antagonists. Circ Res. 1984 Sep;55(3):336–348. doi: 10.1161/01.res.55.3.336. [DOI] [PubMed] [Google Scholar]
  38. Sato R., Noma A., Kurachi Y., Irisawa H. Effects of intracellular acidification on membrane currents in ventricular cells of the guinea pig. Circ Res. 1985 Oct;57(4):553–561. doi: 10.1161/01.res.57.4.553. [DOI] [PubMed] [Google Scholar]
  39. Satoh H., Seyama I. On the mechanism by which changes in extracellular pH affect the electrical activity of the rabbit sino-atrial node. J Physiol. 1986 Dec;381:181–191. doi: 10.1113/jphysiol.1986.sp016321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Taniguchi J., Kokubun S., Noma A., Irisawa H. Spontaneously active cells isolated from the sino-atrial and atrio-ventricular nodes of the rabbit heart. Jpn J Physiol. 1981;31(4):547–558. doi: 10.2170/jjphysiol.31.547. [DOI] [PubMed] [Google Scholar]
  41. Tsien R. W., Bean B. P., Hess P., Lansman J. B., Nilius B., Nowycky M. C. Mechanisms of calcium channel modulation by beta-adrenergic agents and dihydropyridine calcium agonists. J Mol Cell Cardiol. 1986 Jul;18(7):691–710. doi: 10.1016/s0022-2828(86)80941-5. [DOI] [PubMed] [Google Scholar]
  42. Umbach J. A. Changes in intracellular pH affect calcium currents in Paramecium caudatum. Proc R Soc Lond B Biol Sci. 1982 Sep 22;216(1203):209–224. doi: 10.1098/rspb.1982.0071. [DOI] [PubMed] [Google Scholar]
  43. Vogel S., Sperelakis N. Blockade of myocardial slow inward current at low pH. Am J Physiol. 1977 Sep;233(3):C99–103. doi: 10.1152/ajpcell.1977.233.3.C99. [DOI] [PubMed] [Google Scholar]
  44. Yatani A., Goto M. The effect of extracellular low pH on the plateau current in isolated, single rat ventricular cells--a voltage clamp study. Jpn J Physiol. 1983;33(3):403–415. doi: 10.2170/jjphysiol.33.403. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Physiology are provided here courtesy of The Physiological Society

RESOURCES