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Evidence for multiple open states of sodium channels in neuroblastoma cells

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Summary

Open times of voltage-gated sodium channels in neuroblastoma cells were measured during repolarization (following a short depolarizing conditioning pulse) and during moderate depolarization. Conditional and unconditional channel open-time histograms were best fitted by the sum of two exponentials. (The conditional open time was measured from the end of the conditioning pulse until an open channel shuts provided it was open att=0). Time constants of both histograms depended on the postpulse and were shifted to more positive potentials with increasing conditioning pulse potential. This shift could be explained by assuming more than two time constants in the histograms, which could not be separated. Channel open-time histograms from single-pulse experiments showed a maximum att>0. These histograms could be best fitted by an exponential function with three time constants. One term of this function included the difference of two exponentials resulting in a maximum att>0. Open-time histograms showed a definite time dependence. At 2 to 6.5 msec after the beginning of the depolarization the best fit could be obtained by the difference of two exponentials. To these components another term had to be added at 0 to 2 msec. Between 6.5 and 14.0 msec the sum of two exponentials, and after 14.0 msec a single exponential resulted in a good fit. The results support the hypothesis that sodium channels in neuroblastoma cells may have multiple open states. Two of these states are irreversibly coupled.

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References

  • Aldrich, R.W., Corey, D.P., Stevens, C.F. 1983. A reinterpretation of mammalian sodium channel gating based on single channel recording.Nature (London) 306:436–441

    Google Scholar 

  • Armstrong, C.M., Bezanilla, F. 1977. Inactivation of sodium channel. II. Gating current experiments.J. Gen. Physiol. 70:567–590

    PubMed  Google Scholar 

  • Armstrong, C.M., Gilly, W.F. 1979. Fast and slow steps in the activation of sodium channels.J. Gen. Physiol. 74:691–711

    PubMed  Google Scholar 

  • Baumann, G. 1981. Novel kinetics in the sodium conductance system predicted by the aggregation model of channel gating.Biophys. J. 35:699–705

    PubMed  Google Scholar 

  • Baumann, G. 1983. Stochastic modeling of the aggregation-gating site.In: Structure and Function in Excitable Cells. D.C. Chang, I. Tasaki, W.J. Adelman, and H.R. Leuchtag, editors. pp. 255–271. Plenum, New York-London

    Google Scholar 

  • Blatz, A.L., Magleby, K.L. 1986. Correcting single channel data for missed events.Biophys. J. 49:967–980

    PubMed  Google Scholar 

  • Cachelin, A.B., DePeyer, J.E., Kokubun, S., Reuter, H. 1983. Sodium channels in cultured cardiac cells.J. Physiol. (London) 340:389–401

    Google Scholar 

  • Carbone, E., Lux, H.D. 1986. Na channels in cultured chick dorsal root ganglion neurons.Eur. Biophys. J. 13:259–271

    Google Scholar 

  • Chandler, W.K., Meves, H. 1970. Evidence for two types of sodium conductance in axons perfused with sodium fluoride solution.J. Physiol. (London) 211:653–678

    Google Scholar 

  • Colquhoun, D., Hawkes, A.G. 1983. The principles of the stochastic interpretation of ion-channel mechanisms.In: Single Channel Recording. B. Sakmann and E. Neher, editors. pp. 135–175. Plenum, New York

    Google Scholar 

  • Colquhoun, D., Sakmann, B. 1985. Fast events in single-channel currents activated by acetylcholine and its analogues at the frog muscle endplate.J. Physiol. (London) 369:501–557

    Google Scholar 

  • Colquhoun, D., Sigworth, F.J. 1983. Fitting and statistical analysis of single-channel records.In: Single Channel Recording. B. Sakmann and E. Neher, editors. pp. 191–263. Plenum, New York

    Google Scholar 

  • Frankenhaeuser, B., Hodgkin, A.L. 1957. The action of calcium on the electrical properties of squid axons.J. Physiol. (London) 137:218–244

    Google Scholar 

  • French, R.J., Horn, R. 1983. Sodium channel gating: Models, mimics and modifiers.Annu. Rev. Biophys. Bioeng. 12:319–356

    PubMed  Google Scholar 

  • Fukushima, Y. 1981. Identification and kinetic properties of the current through a single Na+ channel.Proc. Natl. Acad. Sci. USA 78:1274–1277

    PubMed  Google Scholar 

  • Gilly, W.F., Armstrong, C.M. 1984. Threshold channels—a novel type of sodium channel in squid giant axon.Nature (London) 309:448–450

    Google Scholar 

  • Goldman, L., Hahin, R. 1978. Initial conditions and the kinetics of the sodium conductance in Myxicola giant axons.J. Gen. Physiol. 72:879–898

    PubMed  Google Scholar 

  • Gration, K.A.F., Lambert, J.J., Ramsey, R.L., Rand, R.P., Usherwood, P.N.R. 1982. Closure of membrane channels gated by glutamate receptors may be a two-step process.Nature (London) 295:599–601

    Google Scholar 

  • Hamill, O.P., Marty, A., Neher, E., Sakmann, B., Sigworth, F.J. 1981. Improved patch clamp techniques for high-resolution current recording from cells and cell-free membrane patches.Pfluegers Arch. Eur. J. Physiol. 391:85–100

    Google Scholar 

  • Hof, D. 1986. A pulse generating and data recording system based on the microcomputer PDP 11/23.Comput. Method. Program. Biomed. 23:309–315

    Google Scholar 

  • Horn, R., Standen, N.B. 1983. Counting kinetic states: The single channel approach.In: The Physiology of Excitable Cells. A. Grinnell and W. Moody, editors. pp. 181–189. A.R. Liss, New York

    Google Scholar 

  • Horn, R., Vandenberg, C.A. 1984. Statistical properties of single sodium channels.J. Gen. Physiol. 83:505–534

    Google Scholar 

  • Horn, R., Vandenberg, C.A., Lange, K. 1984. Statistical analyses of single sodium channels. Effects of N-bromoacetamide.Biophys. J. 45:323–335

    PubMed  Google Scholar 

  • Kiss, T., Nagy, K. 1985. Interaction between sodium channels in mouse neuroblastoma cells.Eur. Biophys. J. 12:13–18

    PubMed  Google Scholar 

  • Kostyuk, P.G., Veselovsky, N.S., Tsyndrenko, A.Y. 1981. Ionic currents in the somatic membrane of rat dorsal root ganglion neurons. I. Sodium currents.Neuroscience 6:2423–2430

    PubMed  Google Scholar 

  • Kunze, P.L., Lacerda, A.E., Wilson, D.L., Brown, A.M. 1985. Cardiac Na currents and the inactivating, reopening, and waiting properties of single cardiac Na channels.J. Gen. Physiol. 86:691–719

    PubMed  Google Scholar 

  • Moolenaar, W.H., Spector, I. 1978. Ionic currents in cultured mouse neuroblastoma cells under voltage-clamp conditions.J. Physiol. (London) 278:265–286

    Google Scholar 

  • Nagy, K. 1987. Subconductance states of single sodium channels modified by chloramine-T and sea anemone toxin in neuroblastoma cells.Eur. Biophys. J. (in press)

  • Nagy, K., Bagany, M. 1986. Multiple discrete single sodium channel current levels in neuroblastoma cells.Proc. Int. Union Physiol. Sci. Vol.XVI p. 441

    Google Scholar 

  • Nagy, K., Kiss, T., Hof, D. 1983. Single Na channels in mouse neuroblastoma cell membrane. Indications for two open states.Pfluegers Arch. Eur. J. Physiol. 399:302–308

    Google Scholar 

  • Neumcke, B., Stämpfli, R. 1983. Alteration of the conductance of Na+ channels in the nodal membrane of frog nerve by holding potential and tetrodotoxin.Biochim. Biophys. Acta 727:177–184

    PubMed  Google Scholar 

  • Patlak, J., Horn, R. 1982. Effect of N-bromoacetamide on single sodium channel currents in excised membrane patches.J. Gen. Physiol. 79:333–351

    Google Scholar 

  • Patlak, J.B., Ortiz, M. 1986. Two modes of gating during late Na+ channel current in frog sartorius muscle.J. Gen. Physiol. 87:305–326

    PubMed  Google Scholar 

  • Patlak, J.B., Ortiz, M., Horn, R. 1986. Open time hetereogeneity during bursting of sodium channels in frog skeletal muscle.Biophys. J. 49:773–777

    PubMed  Google Scholar 

  • Quandt, F.N., Narahashi, T. 1982. Modification of single Na+ channels by batrachotoxin.Proc. Natl. Acad. Sci. USA 79:6732–6736

    PubMed  Google Scholar 

  • Roux, B., Sauvé, R. 1985. A general solution to the time interval omission problem applied to single channel analyses.Biophys. J. 48:149–158

    PubMed  Google Scholar 

  • Schauf, C.L., Bullock, J.O., Pencek, T.L. 1977. Characteristics of sodium tail currents in Myxicola axons. Comparison with membrane asymmetry currents.Biophys. J. 19:7–28

    PubMed  Google Scholar 

  • Sigworth, F.J. 1981. Covariance of nonstationary sodium current fluctuations at the node of Ranvier.Biophys. J. 34:111–133

    PubMed  Google Scholar 

  • Sigworth, F., Neher, E. 1980. Single Na+ channel currents observed in cultured rat muscle cells.Nature (London) 287:447–449

    Google Scholar 

  • Sine, S.M., Steinbach, J.H. 1986. Activation of acetylcholine receptors on clonal mammalian BC3H-1 cells by low concentrations of agonist.J. Physiol. (London) 373:129–162

    Google Scholar 

  • Vandenberg, C.A., Horn, R. 1984. Inactivation viewed through single sodium channels.J. Gen. Physiol. 84:535–564

    PubMed  Google Scholar 

  • Weiss, R.E., Horn, R. 1986. Functional differences between two classes of sodium channels in developing rat skeletal muscle.Science (London) 233:361–364

    Google Scholar 

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Nagy, K. Evidence for multiple open states of sodium channels in neuroblastoma cells. J. Membrain Biol. 96, 251–262 (1987). https://doi.org/10.1007/BF01869307

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