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Measurements of electrical potential differences across yeast plasma membranes with microelectrodes are consistent with values from steady-state distribution of tetraphenylphosphonium inPichia humboldtii

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Summary

Electrical potential differences across the plasma membrane (Δψ) of the yeastPichia humboldtii were measured with microelectrodes (filled with 0.1m KCl) inserted into cells immobilized in microfunnels. The registered Δψ signals were reproducible and stable for several minutes. On attainment of stable reading for Δψ the specific membrane resistanceR sp was determined by applying square-current pulses to the preparation. Both Δψ andR sp were pH dependent and displayed equal but opposite deflection, Δψ reaching its maximal value of −88±9 mV (n=13) andR sp its minimal value of 10 kΩ·cm2 (maximal conductance) at pH 6.5. Uncouplers and the polyene antibiotic nystatin depolarized the cells, decreasing Δψ to −21±15 mV (n=10) with concomitant decrease ofR sp. Comparison of Δψ values from microelectrode measurements with those calculated from the steady-state distribution of tetraphenylphosphonium ions agreed within 10 mV under all physiological conditions tested, except at pH values above 7.0. During microelectrode insertion transient voltage signals (a few msec long) were detected by means of an oscilloscope. These voltage signals were superimposed on the stable Δψ recordings described above. These short voltage signals disappeared in uncoupled cells. The closely related Δψ values obtained by two independent methods (direct measurements with microelectrodes and calculation from steady-state distribution of a lipophilic cation) provide evidence that these values reffect the true membrane potential of intact cells.

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Authors and Affiliations

  1. Institute of Botany, University of Bonn, 5300, Bonn, Federal Republic of Germany

    H. C. Lichtenberg, H. Giebeler & M. Höfer

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  1. H. C. Lichtenberg
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  2. H. Giebeler
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  3. M. Höfer
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Lichtenberg, H.C., Giebeler, H. & Höfer, M. Measurements of electrical potential differences across yeast plasma membranes with microelectrodes are consistent with values from steady-state distribution of tetraphenylphosphonium inPichia humboldtii . J. Membrain Biol. 103, 255–261 (1988). https://doi.org/10.1007/BF01993985

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  • DOI: https://doi.org/10.1007/BF01993985

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