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. 1987 Feb 1;89(2):253–274. doi: 10.1085/jgp.89.2.253

Gating of Na channels. Inactivation modifiers discriminate among models

PMCID: PMC2215892  PMID: 2435840

Abstract

Macroscopic Na currents were recorded from N18 neuroblastoma cells by the whole-cell voltage-clamp technique. Inactivation of the Na currents was removed by intracellular application of proteolytic enzymes, trypsin, alpha-chymotrypsin, papain, or ficin, or bath application of N- bromoacetamide. Unlike what has been reported in squid giant axons and frog skeletal muscle fibers, these treatments often increased Na currents at all test pulse potentials. In addition, removal of inactivation gating shifted the midpoint of the peak Na conductance- voltage curve in the negative direction by 26 mV on average and greatly prolonged the rising phase of Na currents for small depolarizations. Polypeptide toxins from Leiurus quinquestriatus scorpion and Goniopora coral, which slow inactivation in adult nerve and muscle cells, also increase the peak Na conductance and shift the peak conductance curve in the negative direction by 7-10 mV in neuroblastoma cells. Control experiments argue against ascribing the shifts to series resistance artifacts or to spontaneous changes of the voltage dependence of Na channel kinetics. The negative shift of the peak conductance curve, the increase of peak Na currents, and the prolongation of the rise at small depolarization after removal of inactivation are consistent with gating kinetic models for neuroblastoma cell Na channels, where inactivation follows nearly irreversible activation with a relatively high, voltage- independent rate constant and Na channels open only once in a depolarization. As the same kind of experiment does not give apparent shifting of activation and prolongation of the rising phase of Na currents in adult axon and muscle membranes, the Na channels of these other membranes probably open more than once in a depolarization.

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Selected References

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  1. Aldrich R. W., Corey D. P., Stevens C. F. A reinterpretation of mammalian sodium channel gating based on single channel recording. Nature. 1983 Dec 1;306(5942):436–441. doi: 10.1038/306436a0. [DOI] [PubMed] [Google Scholar]
  2. Aldrich R. W., Stevens C. F. Inactivation of open and closed sodium channels determined separately. Cold Spring Harb Symp Quant Biol. 1983;48(Pt 1):147–153. doi: 10.1101/sqb.1983.048.01.017. [DOI] [PubMed] [Google Scholar]
  3. Armstrong C. M., Bezanilla F. Inactivation of the sodium channel. II. Gating current experiments. J Gen Physiol. 1977 Nov;70(5):567–590. doi: 10.1085/jgp.70.5.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Armstrong C. M., Bezanilla F., Rojas E. Destruction of sodium conductance inactivation in squid axons perfused with pronase. J Gen Physiol. 1973 Oct;62(4):375–391. doi: 10.1085/jgp.62.4.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Armstrong C. M., Gilly W. F. Fast and slow steps in the activation of sodium channels. J Gen Physiol. 1979 Dec;74(6):691–711. doi: 10.1085/jgp.74.6.691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Armstrong C. M. Sodium channels and gating currents. Physiol Rev. 1981 Jul;61(3):644–683. doi: 10.1152/physrev.1981.61.3.644. [DOI] [PubMed] [Google Scholar]
  7. Attwell D., Cohen I., Eisner D., Ohba M., Ojeda C. The steady state TTX-sensitive ("window") sodium current in cardiac Purkinje fibres. Pflugers Arch. 1979 Mar 16;379(2):137–142. doi: 10.1007/BF00586939. [DOI] [PubMed] [Google Scholar]
  8. Bergman C., Dubois J. M., Rojas E., Rathmayer W. Decreased rate of sodium conductance inactivation in the node of Ranvier induced by a polypeptide toxin from sea anemone. Biochim Biophys Acta. 1976 Nov 11;455(1):173–184. doi: 10.1016/0005-2736(76)90162-0. [DOI] [PubMed] [Google Scholar]
  9. Bezanilla F., Armstrong C. M. Inactivation of the sodium channel. I. Sodium current experiments. J Gen Physiol. 1977 Nov;70(5):549–566. doi: 10.1085/jgp.70.5.549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Catterall W. A. Activation of the action potential Na+ ionophore of cultured neuroblastoma cells by veratridine and batrachotoxin. J Biol Chem. 1975 Jun 10;250(11):4053–4059. [PubMed] [Google Scholar]
  11. Catterall W. A. Binding of scorpion toxin to receptor sites associated with sodium channels in frog muscle. Correlation of voltage-dependent binding with activation. J Gen Physiol. 1979 Sep;74(3):375–391. doi: 10.1085/jgp.74.3.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Catterall W. A. Neurotoxins that act on voltage-sensitive sodium channels in excitable membranes. Annu Rev Pharmacol Toxicol. 1980;20:15–43. doi: 10.1146/annurev.pa.20.040180.000311. [DOI] [PubMed] [Google Scholar]
  13. Catterall W. A. Purification of a toxic protein from scorpion venom which activates the action potential Na+ ionophore. J Biol Chem. 1976 Sep 25;251(18):5528–5536. [PubMed] [Google Scholar]
  14. Catterall W. A., Ray R., Morrow C. S. Membrane potential dependent binding of scorpion toxin to action potential Na+ ionophore. Proc Natl Acad Sci U S A. 1976 Aug;73(8):2682–2686. doi: 10.1073/pnas.73.8.2682. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fernandez J. M., Fox A. P., Krasne S. Membrane patches and whole-cell membranes: a comparison of electrical properties in rat clonal pituitary (GH3) cells. J Physiol. 1984 Nov;356:565–585. doi: 10.1113/jphysiol.1984.sp015483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gilly W. F., Armstrong C. M. Slowing of sodium channel opening kinetics in squid axon by extracellular zinc. J Gen Physiol. 1982 Jun;79(6):935–964. doi: 10.1085/jgp.79.6.935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gilly W. F., Armstrong C. M. Threshold channels--a novel type of sodium channel in squid giant axon. 1984 May 31-Jun 6Nature. 309(5967):448–450. doi: 10.1038/309448a0. [DOI] [PubMed] [Google Scholar]
  18. Goldman L. Kinetics of channel gating in excitable membranes. Q Rev Biophys. 1976 Nov;9(4):491–526. doi: 10.1017/s0033583500002651. [DOI] [PubMed] [Google Scholar]
  19. Gonoi T., Ashida K., Feller D., Schmidt J., Fujiwara M., Catterall W. A. Mechanism of action of a polypeptide neurotoxin from the coral Goniopora on sodium channels in mouse neuroblastoma cells. Mol Pharmacol. 1986 Apr;29(4):347–354. [PubMed] [Google Scholar]
  20. Gonoi T., Hille B., Catterall W. A. Voltage clamp analysis of sodium channels in normal and scorpion toxin-resistant neuroblastoma cells. J Neurosci. 1984 Nov;4(11):2836–2842. doi: 10.1523/JNEUROSCI.04-11-02836.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gonoi T., Sherman S. J., Catterall W. A. Voltage clamp analysis of tetrodotoxin-sensitive and -insensitive sodium channels in rat muscle cells developing in vitro. J Neurosci. 1985 Sep;5(9):2559–2564. doi: 10.1523/JNEUROSCI.05-09-02559.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. HODGKIN A. L., HUXLEY A. F. A quantitative description of membrane current and its application to conduction and excitation in nerve. J Physiol. 1952 Aug;117(4):500–544. doi: 10.1113/jphysiol.1952.sp004764. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Hille B. Gating in sodium channels of nerve. Annu Rev Physiol. 1976;38:139–152. doi: 10.1146/annurev.ph.38.030176.001035. [DOI] [PubMed] [Google Scholar]
  25. Horn R., Vandenberg C. A., Lange K. Statistical analysis of single sodium channels. Effects of N-bromoacetamide. Biophys J. 1984 Jan;45(1):323–335. doi: 10.1016/S0006-3495(84)84158-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Horn R., Vandenberg C. A. Statistical properties of single sodium channels. J Gen Physiol. 1984 Oct;84(4):505–534. doi: 10.1085/jgp.84.4.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Keynes R. D. The Croonian Lecture, 1983. Voltage-gated ion channels in the nerve membrane. Proc R Soc Lond B Biol Sci. 1983 Nov 22;220(1218):1–30. doi: 10.1098/rspb.1983.0086. [DOI] [PubMed] [Google Scholar]
  28. Khodorov B. I. Sodium inactivation and drug-induced immobilization of the gating charge in nerve membrane. Prog Biophys Mol Biol. 1981;37(2):49–89. doi: 10.1016/0079-6107(82)90020-7. [DOI] [PubMed] [Google Scholar]
  29. Koppenhöfer E., Schmidt H. Die Wirkung von Skorpiongift auf die Ionenströme des Ranvierschen Schnürrings. II. Unvollständiage Natrium-Inaktivierung. Pflugers Arch. 1968;303(2):150–161. doi: 10.1007/BF00592632. [DOI] [PubMed] [Google Scholar]
  30. Matteson D. R., Armstrong C. M. Evidence for a population of sleepy sodium channels in squid axon at low temperature. J Gen Physiol. 1982 May;79(5):739–758. doi: 10.1085/jgp.79.5.739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Nonner W. Relations between the inactivation of sodium channels and the immobilization of gating charge in frog myelinated nerve. J Physiol. 1980 Feb;299:573–603. doi: 10.1113/jphysiol.1980.sp013143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Nonner W., Spalding B. C., Hille B. Low intracellular pH and chemical agents slow inactivation gating in sodium channels of muscle. Nature. 1980 Mar 27;284(5754):360–363. doi: 10.1038/284360a0. [DOI] [PubMed] [Google Scholar]
  33. Oxford G. S. Some kinetic and steady-state properties of sodium channels after removal of inactivation. J Gen Physiol. 1981 Jan;77(1):1–22. doi: 10.1085/jgp.77.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Oxford G. S., Wu C. H., Narahashi T. Removal of sodium channel inactivation in squid giant axons by n-bromoacetamide. J Gen Physiol. 1978 Mar;71(3):227–247. doi: 10.1085/jgp.71.3.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Patlak J. B., Ortiz M. Two modes of gating during late Na+ channel currents in frog sartorius muscle. J Gen Physiol. 1986 Feb;87(2):305–326. doi: 10.1085/jgp.87.2.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Patlak J., Horn R. Effect of N-bromoacetamide on single sodium channel currents in excised membrane patches. J Gen Physiol. 1982 Mar;79(3):333–351. doi: 10.1085/jgp.79.3.333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rojas E., Rudy B. Destruction of the sodium conductance inactivation by a specific protease in perfused nerve fibres from Loligo. J Physiol. 1976 Nov;262(2):501–531. doi: 10.1113/jphysiol.1976.sp011608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Sevcik C., Narahashi T. Effects of proteolytic enzymes on ionic conductances of squid axon membranes. J Membr Biol. 1975 Dec 4;24(3-4):329–339. doi: 10.1007/BF01868630. [DOI] [PubMed] [Google Scholar]
  39. Shrager P. Specific chemical groups involved in the control of ionic conductance in nerve. Ann N Y Acad Sci. 1975 Dec 30;264:293–303. doi: 10.1111/j.1749-6632.1975.tb31490.x. [DOI] [PubMed] [Google Scholar]
  40. Sigworth F. J. Covariance of nonstationary sodium current fluctuations at the node of Ranvier. Biophys J. 1981 Apr;34(1):111–133. doi: 10.1016/S0006-3495(81)84840-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Starkus J. G., Shrager P. Modification of slow sodium inactivation in nerve after internal perfusion with trypsin. Am J Physiol. 1978 Nov;235(5):C238–C244. doi: 10.1152/ajpcell.1978.235.5.C238. [DOI] [PubMed] [Google Scholar]
  42. Stimers J. R., Bezanilla F., Taylor R. E. Sodium channel activation in the squid giant axon. Steady state properties. J Gen Physiol. 1985 Jan;85(1):65–82. doi: 10.1085/jgp.85.1.65. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. TASAKI I., TAKENAKA T. EFFECTS OF VARIOUS POTASSIUM SALTS AND PROTEASES UPON EXCITABILITY OF INTRACELLULARLY PERFUSED SQUID GIANT AXONS. Proc Natl Acad Sci U S A. 1964 Sep;52:804–810. doi: 10.1073/pnas.52.3.804. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Vandenberg C. A., Horn R. Inactivation viewed through single sodium channels. J Gen Physiol. 1984 Oct;84(4):535–564. doi: 10.1085/jgp.84.4.535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Wang G. K., Strichartz G. Kinetic analysis of the action of Leiurus scorpion alpha-toxin on ionic currents in myelinated nerve. J Gen Physiol. 1985 Nov;86(5):739–762. doi: 10.1085/jgp.86.5.739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Warashina A., Fujita S. Effect of sea anemone toxins on the sodium inactivation process in crayfish axons. J Gen Physiol. 1983 Mar;81(3):305–323. doi: 10.1085/jgp.81.3.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Zubov A. N. Nekotorye svoistva natrievykh kanalov membrany kul'tiviruemykh kletok neiroblastomy klona N18 A-1. Tsitologiia. 1980 Oct;22(10):1207–1213. [PubMed] [Google Scholar]

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