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Determination of the electromotive force of active sodium transport in frog skin epithelium (Rana temporaria) from presteady-state flux ratio experiments

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Summary

The presteady-state influxes and effluxes of sodium across frog skin epithelium have been determined as a function of time while all electrophysiological parameters were maintained constant. The fluxes measured were resolved in the fractions which have passed a pathway through the cells and those that have used a paracellular pathway. The procedure is based on the theory that all presteady-state flux ratios have to be equal to the steady-state flux ratio if only one pathway is involved. The flux ratios for the transcellular route were used to calculate the electromotive force of the sodium pump. The calculation hinges on the assumptions (a) that both influx and efflux have to pass through the sodium pump and (b) that single file diffusion of sodium is not taking place anywhere along the path. The validity of both assumptions is discussed. Our calculated values for the electromotive force of the sodium pumpE aNa vary between 146 and 200 mV, which is in agreement with the energy of the ATP/ADP system. There is a distinct indication that, as the electrochemical gradient for sodium opposing the transport is being increased, the emf increases towards an asymptotic value around 200 mV. The relation between the value ofE aNa and the cellular phosphorylation potential for ATP is discussed.

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References

  • Benos, D.J., Hyde, B.A., Latorre, R., 1983. Sodium flux ratio through the amiloride-sensitive entry pathway in frog skin.J. Gen. Physiol. 81:667–685

    Google Scholar 

  • Biber, T.U.L., Mullen, T.L. 1977. Effect of inhibitors on transepithelial efflux of Na and nonelectrolytes in frog skin.Am. J. Physiol. 232:C67-C75

    Google Scholar 

  • Canessa, M., Labarca, P., DiBona, D.R., Leaf, A. 1978. Energetics of sodium transport in toad urinary bladder.Proc. Natl. Acad. Sci. USA 75:4591–4595

    Google Scholar 

  • Chase, H.S., Al-Awqati, Q. 1981. Regulation of the sodium permeability of the luminal border of toad bladder by intracellular sodium and calcium.J. Gen. Physiol. 77:693–712

    Google Scholar 

  • Eskesen, K., Lim, J.J., Ussing, H.H. 1985. Evaluation of transport pathways for Na+ across frog skin epithelium by means of presteady state flux ratio.J. Membrane Biol. 86:99–104

    Google Scholar 

  • Ferreira, K.T.G., Hill, S.B. 1978. Chloride dependence of active sodium transport in frog skin: The role of intercellular spaces.J. Physiol. (London) 283:283–305

    Google Scholar 

  • Fuchs, W., Hviid Larsen, E., Lindemann, B., 1977. Current-voltage curve of sodium channels and concentration dependence of sodium permeability in frog skin.J. Physiol. (London) 267:137–166

    Google Scholar 

  • Garrahan, P.J., Glynn, I.M. 1967a. The incorporation of inorganic phosphate into adenosine triphosphate by reversal of the sodium pump.J. Physiol. (London) 192:237–256

    Google Scholar 

  • Garrahan, P.J., Glynn, I.M. 1967b. The behaviour of the sodium pump in red cells in the absence of external potassium.J. Physiol. (London) 192:159–174

    Google Scholar 

  • Garrahan, P.J., Glynn, I.M. 1967c. Factors affecting the relative magnitudes of the sodium: potassium and sodium: sodium exchanges catalysed by the sodium pump.J. Physiol., (London) 192:189–216

    Google Scholar 

  • Glynn, I.M., Lew, V.L., 1970. Synthesis of adenosine triphosphate at the expense of downhill cation movements in intact human red cells.J. Physiol. (London) 207:393–402

    Google Scholar 

  • Harvey, B.J., Kernan, R.P., 1984. Intracellular ion activities in frog skin in relation to external sodium and effects of amiloride and/or ouabain.J. Physiol. (London) 349:501–517

    Google Scholar 

  • Helman, S.I., Fisher, R.S. 1977. Microelectrode studies of the active Na transport pathway of frog skin.J. Gen. Physiol. 69:571–604

    Google Scholar 

  • Hodgkin, A.L., Keynes, R.D. 1955. The potassium permeability of a giant nerve fibre.J. Physiol. (London) 128:61–88

    Google Scholar 

  • Holian, A., Owen, A.S., Wilson, D.F. 1977. Control of respiration in isolated mitochondria: Quantitative evaluation of the dependence of respiratory rates on (ATP). (ADP), and (P i ).Arch. Biochem. Biophys. 181:164–171

    Google Scholar 

  • Koefoed-Johnsen, V., Ussing, H.H. 1958. The nature of the frog skin potential.Acta. Physiol. Scand. 42:298–308

    Google Scholar 

  • Kristensen, P. 1978. Effect of amiloride on chloride transport across amphibian epithelia.J. Membrane Biol. Special Issue:167–185

    Google Scholar 

  • Larsen, E.H. 1972. Effect of amiloride, cyanide and ouabain on the active transport pathway in toad skin.In: Transport Mechanisms in Epithelia. pp. 131–143. Munksgaard. Copenhagen

    Google Scholar 

  • Leaf, A., Renshaw, A. 1957. Ion transport and respiration of isolated frog skin.Biochem. J.,65:82–90

    Google Scholar 

  • Lindemann, B., Van Driessche W. 1978. The mechanism of Na uptake through Na-selective channels in the epithelium of frog skin.In: Membrane Transport Processes 1. J.F. Hoffman, editor. pp. 155–178. Raven. New York

    Google Scholar 

  • Nagel, W. 1976. The intracellular electrical potential profile of the frog skin epithelium.Pfluegers Arch. 365:135–143

    Google Scholar 

  • Nagel, W., Garcia-Diaz, J.F., Armstrong, W.McD. 1981. Intracellular ionic activities in frog skin.J. Membrane Biol. 61:127–134

    Google Scholar 

  • Nielsen, R. 1979. A 3 to 2 coupling of the Na−K pump responsible for the transepithelial Na transport in frog skin disclosed by the effect of Ba.Acta Physiol. Scand. 107:189–191

    Google Scholar 

  • Nielsen, R. 1984. Active transepithelial potassium transport in frog skin via specific potassium channels in the apical membrane.Acta Physiol. Scand. 120:287–296

    Google Scholar 

  • Rick, R., Dörge, A., E. von Arnim, Thurau, K. 1978. Electron microprobe analysis of frog skin epithelium: Evidence for a syncytial sodium transport compartment.J. Membrane Biol. 39:313–331

    Google Scholar 

  • Rick, R., Roloff, C., Dörge, A., Beck, F.X., Thurau, K. 1984. Intracellular electrolyte concentrations in the frog skin epithelium: Effect of vasopressin and dependence on the Na concentration in the bathing media.J. Membrane Biol. 78:129–145

    Google Scholar 

  • Sten-Knudsen, O., Ussing, H.H. 1981. The flux ratio equation under nonstationary conditions.J. Membrane Biol. 63:233–242

    Google Scholar 

  • Taylor, A., Windhager, E.E. 1979. Possible role of cytosolic calcium and Na−Ca exchange in regulation of transepithelial sodium transport.Am. J. Physiol. 236:F505-F512

    Google Scholar 

  • Tosteson, D.C., Hoffman, J.F. 1960. Regulation of cell volume by active cation transport in high and low potassium sheep red cells.J. Gen. Physiol. 44:169–194

    Google Scholar 

  • Ussing, H.H. 1982. Volume regulation of frog skin epithelium.Acta Physiol. Scand. 114:363–369

    Google Scholar 

  • Ussing, H.H., Eskesen, K., Lim, J. 1981. The flux ratio transient as a tool for separating transport pathways in epithelia.In: Epithelial Ion and Water Transport. A.D.C. Macknight and J.P. Leader editor Raven, New York

    Google Scholar 

  • Ussing, H.H., Kruhøffer, P., Hess-Thaysen, J., Thorn, N.A. 1960. The alkali metal ions in biology.In: Handbuch der experimentellen Pharmakologie Band 13. pp. 112–129. O. Eichler and A. Farah, editors. Springer-Verlag. Berlin

    Google Scholar 

  • Ussing, H.H., Windhager, E.E., 1964. Nature of shunt path and active sodium transport path through frog skin epithelium.Acta Physiol. Scand. 61:484–504

    Google Scholar 

  • Ussing, H.H., Zerahn, K. 1951. Active transport of sodium as the source of electric current in the short-circuited isolated frog skin.Acta Physiol. Scand. 23:110–127

    Google Scholar 

  • Veech, R.L., Lawson, J.W.R., Cornell, N.W., Krebs, H.A. 1979. Cytosolic phosphorylation potential.J. Biol. Chem. 251:6538–6547

    Google Scholar 

  • Wilson, D.F., Owen, C., Mela, L., Weiner, L. 1973. Control of mitochondrial respiration by the phosphate potential.Biochem. Biophys. Res. Commun. 53:326–333

    Google Scholar 

  • Zeiske, W., Lindemann, B. 1974. Chemical stimulation of Na+ current through the outer surface of frog skin epithelium.Biochim. Biophys. Acta 352:323–326

    Google Scholar 

  • Zerahn, K. 1956. Oxygen consumption and active sodium transport in the isolated and short-circuited frog skin.Acta Physiol. Scand. 36:301–318

    Google Scholar 

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Eskesen, K., Ussing, H.H. Determination of the electromotive force of active sodium transport in frog skin epithelium (Rana temporaria) from presteady-state flux ratio experiments. J. Membrain Biol. 86, 105–111 (1985). https://doi.org/10.1007/BF01870777

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