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The quantal gating charge of sodium channel inactivation

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Abstract

Using a very low noise voltage clamp technique it has been possible to record from the squid giant axon a slow component of gating current (I g ) during the inactivation phase of the macroscopic sodium current (I Na ) which was hitherto buried in the baseline noise. In order to examine whether this slowI g contains gating charge that originates from transitions between the open (O) and the inactivated (I) states, which would indicate a true voltage dependence of inactivation, or whether other transitions contribute charge to slowI g , a new model independent analysis termed isochronic plot analysis has been developed. From a direct correlation ofI g and the time derivative of the sodium conductance dg Na/d the condition when only O-I transitions occur is detected. Then the ratio of the two signals is constant and a straight line appears in an isochronic plot ofI g vs. dg Na/d . Its slope does not depend on voltage or time and corresponds to the quantal gating charge of the O-I transition (q h ) divided by the single channel ionic conductance (γ). This condition was found at voltages above − 10 mV up to + 40 mV and a figure of 1.21e − was obtained forq h at temperatures of 5 and 15°C. At lower voltages additional charge from other transitions, e.g. closed to open, is displaced during macroscopic inactivation. This means that conventional Eyring rate analysis of the inactivation time constant τ h is only valid above − 10 mV and here the figure forq h was confirmed also from this analysis. It is further shown that most of the present controversies surrounding the voltage dependence of inactivation can be clarified. The validity of the isochronic plot analysis has been confirmed using simulated gating and ionic currents.

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Abbreviations

I g :

gating current

I Na :

sodium ionic current

g Na :

macroscopic sodium conductance

γ:

single channel conductance

C, O, I:

closed, open, inactivated state occupancy of channels

g h :

quantal charge displaced in a single O-I transition of Na channel

e − :

equivalent electron charge

h :

index referring to inactivation process

S l :

limiting slope in isochronic plot see Eq.(3)

δ:

fractional distance, see Fig. 4 and (4, 5)

TMA:

tetramethylammonium

TTX:

tetrodotoxin

Tris:

tris(hydroxymethyl)aminomethane

HEPES:

N-2-hydroxyethylpiperazine-N′-2-ethanesulfonic acid

References

  • Aldrich RW, Stevens CF (1987) Voltage-depenent gating of single sodium channels from mammalian neuroblastoma cells. J Neurosci 7:418–431

    Google Scholar 

  • Aldrich RW, Corey DP, Stevens CF (1983) A reinterpretation of mammalian sodium channel gating based on single channel recording. Nature (London) 306:436–441

    Google Scholar 

  • Almers W, Armstrong CM (1980) Survival of K− permeability and gating currents in squid axons perfused with K+-free media. J Gen Physiol 75:61–87

    Google Scholar 

  • Armstrong CM (1981) Sodium channels and gating currents. Physiol Rev 61:644–683

    Google Scholar 

  • Armstrong CM, Bezanilla F (1977) Inactivation of the sodium channel. II. Gating current experiments. J Gen Physiol 70:567–590

    Google Scholar 

  • Augustine CK, Bezanilla F (1990) Phosphorylation modulates potassium conductance and gating current of perfused giant axons of squid. J Gen Physiol 95:245–271

    Google Scholar 

  • Bekkers JM, Greeff NG, Keynes RD (1986a) The conductance and density of sodium channels in the cut-open squid giant axon. J Physiol (London) 377:463–486

    Google Scholar 

  • Bekkers JM, Forster IC, Greeff NG, Keynes RD (1986b) Improvements in the recording of gating currents in the squid giant axon. J Physiol (London) 381:9P

    Google Scholar 

  • Bekkers JM, Forster IC, Greeff NG (1990) Gating current associated with inactivated states of the squid axon sodium channel. Proc Natl Acad Sci USA 87:8311–8315

    Google Scholar 

  • Berman MF, Camardo SS, Robinson RB, Siegelbaum SA (1989) Single sodium channels from canine ventricular myocytes: Voltages dependence and relative rates of activation and inactivation. J Physiol (London) 415:503–531

    Google Scholar 

  • Bezanilla F (1985) Gating of sodium and potassium channels. J Membr Biol 88:97–112

    Google Scholar 

  • Bezanilla F (1987) Single sodium channels from the squid giant axon. Biophys J 52:1087–1090

    Google Scholar 

  • Bullock JO, Schauf CL (1978) Combined voltage-clamp and dialysis of Myxicola axons: behaviour of membrane asymmetry currents. J Physiol (London) 278:309–324

    Google Scholar 

  • Catterall WA (1988) Structure and function of voltage-sensitive ion channels. Science 242:50–61

    Google Scholar 

  • Chandler WK, Meves H (1970a) Sodium and potassium currents in squid axons perfused with fluoride solutions. J Physiol (London) 211:623–652

    Google Scholar 

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

    Google Scholar 

  • Conti F, Stühmer W (1989) Quantal charge redistributions accompanying the structural transitions of sodium channels. Eur Biophys J 17:53–59

    Google Scholar 

  • Cota G, Armstrong CM (1989) Sodium channel gating in a clonal pituitry cells. The inactivation step is not voltage dependent. J Gen Physiol 94:213–232

    Google Scholar 

  • Forster IC, Greeff NG (1989) The quantal gating charge of sodium channel inactivation in the squid giant axon (L. forbesii). J Physiol (London) 418:11P

    Google Scholar 

  • Forster IC, Greeff NG (1990) High resolution recording of asymmetry currents from the squid giant axon-technical aspects of voltage clamp design. J Neurosci Methods 33:185–205

    Google Scholar 

  • Gilly WF, Armstrong CM (1980) Gating current and potassium channels in the squid giant axon of the squid. Biophys J 29:485–492

    Google Scholar 

  • Gonoi T, Hille B (1987) Gating of sodium channels. Inactivation modifiers discriminate among models. J Gen Physiol 89:253–274

    Google Scholar 

  • Greeff NG, Keynes RD, van Helden DF (1982) Fractionation of the asymmetry current in the squid giant axon into inactivating and non-inactivating components. Proc R Soc B215:375–389

    Google Scholar 

  • Greeff NG, Forster IC (1990) The quantal gating charge of Na channel inactivation obtained from gating currents and ionic current rate analysis. Biophys J 57:103a

    Google Scholar 

  • Guy HR (1988) A model relating the structure of the sodium channel to its function. Current Topics Membrane Transport 3:289–308

    Google Scholar 

  • Guy HR, Seetharamulu P (1986) Molecular model of the action potential sodium channel. Proc Natl Acad Sci USA 83: 508–512

    Google Scholar 

  • Hille B (1984) Ionic channels of excitable membranes. Sinauer, Sunderland, Mass

    Google Scholar 

  • Hodgkin AL, Huxley AF (1952) A quantitative description of membrane current and its application to conduction and excitation in nerve. J Physiol (London) 117:500–544

    Google Scholar 

  • Horn R, Vandenberg CA, Lange K (1984) Statistical analysis of single sodium channels. Effects of N-Bromoacetamide. Biophys J 45:323–355

    Google Scholar 

  • Hoshi T, Zagotta WN, Aldrich RW (1990) Biophysical and molecular mechanism ofShaker potassium channel inactivation. Science 250:533–538

    Google Scholar 

  • Keynes RD (1991) On the voltage dependence of inactivation in the sodium channel of the squid giant axon. Proc Soc B 243:47–53

    Google Scholar 

  • Keynes RD, Rojas E (1976) Kinetics and steady state relationships between activation of the sodium conductance and movement of the gating particles in the squid axons. J Physiol (London) 255:157–189

    Google Scholar 

  • Keynes RD, Greeff NG, Forster IC (1990) Kinetic analysis of the sodium gating current in the squid giant axon. Proc Soc B 240:411–423

    Google Scholar 

  • Kosower EM (1985) A structural and dynamic molecular model for the sodium channel ofElectrophorus electricus. FEBS Lett 182:234–242

    Google Scholar 

  • Landowne D (1990) Chloramine-T alters the nerve membrane hirefringence response. J Membrane Biol 113:123–129

    Google Scholar 

  • Meves H, Pohl J-A (1990) A slow component in the gating current of the frog node of Ranvier. Pflügers Arch 416:162–169

    Google Scholar 

  • Nagy K, Kiss T, Hof D (1983) Single Na channels in mouse neuroblastoma cell membrane. — Indications for two open states. Pflügers Arch 399:302–308

    Google Scholar 

  • Noda M, Shimizu S, Tanabe T, Takai T, Kayano T, Ikeda T, Takahashi H, Nakayama H, Kanaoka Y, Minamino N, Kangawa K, Matsuo H, Raftery MA, Hirose T, Inayama S, Hayashida H, Miyata T, Numa S (1984) Primary structure ofElectrophorus electricus sodium channel deduced from cDNA sequence. Nature 312:121–127

    Google Scholar 

  • Noda M, Ikeda T, Kayano T, Suzuki H, Takeshima H, Kurasaki H, Takahashi H, Numa S (1986) Existence of distinct sodium channels from cloned cDNA. Nature 320:188–192

    Google Scholar 

  • Rojas E, Keynes RD (1975) On the relation between displacement currents and activation of the sodium conductance in the squid giant axon. Phil Trans R Soc London B 270:459–482

    Google Scholar 

  • Salkoff L, Butler A, Wei A, Scavarda N, Giffen K, Ifune C, Goodman R, Mandel G (1987a) Genomic organization and deduced amino acid sequence of a putative sodium channel gene inDrosophila. Science 237:744–749

    Google Scholar 

  • Salkoff L, Butler A, Wei A, Scavarda N, Baker K, Pauron D, Smith C (1987b) Molecular biology of the voltage-gated sodium channel. Trends Neurosci 10:522–527

    Google Scholar 

  • Scanley BE, Hanck DA, Chay T, Fozzard HA (1990) Kinetic analysis of single sodium channels from canine cardiac Purkinje cells. J Gen Physiol 95:411–437

    Google Scholar 

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

    Google Scholar 

  • Spires S, Begenisich T (1989) Pharmacological and kinetic analysis of K channel gating currents. J Gen Physiol 93:263–283

    Google Scholar 

  • Stevens CF (1987) Sodium channel structure-function relations. In: Hille B, Fambrough DM (eds) Proteins in excitable membranes. Volume of The Society of General Physiology Series 41:99–108

  • Stimers JR, Bezanilla F, Taylor RE (1985) Sodium channel activation in the squid giant axon. Steady state properties. J Gen Physiol 85:65–82

    Google Scholar 

  • Stühmer W, Conti F, Suzuki H, Wang X, Noda M, Yahagi N, Kubo H, Numa S (1989) Structural parts involved in activation and inactivation of the sodium channel. Nature 339:597–603

    Google Scholar 

  • Swenson RP (1983) A slow component of gating current in crayfish giant axons resembles inactivation charge movement. Biophys J 41:245–249

    Google Scholar 

  • Tejedor FJ, Catterall WA (1988) Site of covalent attachment of α-scorpion toxin derivatives in domain I of the sodium channel subunit. Proc Natl Acad Sci USA 85:8742–8746

    Google Scholar 

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

    Google Scholar 

  • White MM, Bezanilla F (1985) Activation of squid axon K+ channels: Ionic and gating current studies. J Gen Physiol 85:539–554

    Google Scholar 

  • Yamamoto D, Yeh JZ, Narahashi T (1984) Voltage-dependent calcium block of normal and tetramethrin-modified single sodium channels. Biophys J 45:337–344

    Google Scholar 

  • Yue DT, Lawrence JH, Marban E (1989) Two molecular transitions influence cardiac sodium channel gating. Science 244:349–352

    Google Scholar 

Download references

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Authors and Affiliations

  1. Physiologisches Institut, Universität Zürich-Irchel, Winterthurer Strasse 190, CH-8057, Zurich, Switzerland

    N. G. Greeff & I. C. Forster

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  1. N. G. Greeff
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  2. I. C. Forster
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Greeff, N.G., Forster, I.C. The quantal gating charge of sodium channel inactivation. Eur Biophys J 20, 165–176 (1991). https://doi.org/10.1007/BF01561139

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