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External Ni2+ and ENaC in A6 Cells: Na+ Current Stimulation by Competition at a Binding Site for Amiloride and Na+

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Abstract

In cultured A6 monolayers from distal Xenopus kidney, external Ni2+ stimulated active Na+ uptake via the epithelial Na+ channel, ENaC. Transepithelial capacitance measurements ruled out exocytosis of ENaC-containing vesicles underlying the Ni2+ effect. Na+ current noise analysis was performed using the neutral Na+-channel blocker 6-chloro-3,5-diamino-pyrazine-2-carboxamide (CDPC) and amiloride. The analysis of CDPC-induced noise in terms of a three-state channel model revealed that Ni2+ elicits an increase in the number of open channels as well as in the spontaneous open probability. While Ni2+ had no influence on CDPC-blocker kinetics, the macroscopic and microscopic blocking kinetics of amiloride were affected. Ni2+ turned out to compete with amiloride for a putative binding site but not with CDPC. Moreover, external Na+—known to compete with amiloride and so producing the “self-inhibition” phenomenon—and Ni2+ exerted mutually exclusive analogous effects on amiloride kinetics. Na+ current kinetics revealed that Ni2+ prevents ENaC to be downregulated by self-inhibition. Co2+ behaved similarly to Ni2+, whereas Zn2+ did not. Attempts to disclose the chemical nature of the site reacting with Ni2+ suggested cysteine but not histidine as reaction partner.

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Acknowledgements

This project was supported by research grants from the ‘Fonds voor wetenschappelijk onderzoek-Vlaanderen' (G.0179.99 and G.0277.03), the Interuniversity Poles of Attraction Program-Belgian State, Prime Minister's Office-Federal Office for Scientific, Technical, and Cultural Affairs IUAP P4/23. D. Cucu was supported by the Research Council scholarships program of the K.U.Leuven.

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  1. Laboratory of Physiology, K. U. Leuven, Campus Gasthuisberg O/N, B-3000 LEUVEN, Belgium

    D. Cucu, J. Simaels & W. Van Driessche

  2. Institut für Zoophysiologie, Universität Osnabrück, Barbarastrasse, 11, D-49069 Osnabrück, Germany

    W. Zeiske

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  1. D. Cucu
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  2. J. Simaels
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Correspondence to W. Van Driessche.

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Cucu, D., Simaels, J., Van Driessche, W. et al. External Ni2+ and ENaC in A6 Cells: Na+ Current Stimulation by Competition at a Binding Site for Amiloride and Na+ . J. Membrane Biol. 194, 33–45 (2003). https://doi.org/10.1007/s00232-003-2023-y

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