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Na+/H+-exchange in A6 cells: Polarity and vasopressin regulation

The Journal of Membrane Biology volume 130, pages 105–114 (1992)Cite this article

Summary

We have analyzed the mechanism of Na+-dependent pHi; recovery from an acid load in A6 cells (an amphibian distal nephron cell line) by using the intracellular pH indicator 2′7′-bis(2-carboxyethyl)5, 6 carboxyfluorescein (BCECF) and single cell microspectrofluorometry. A6 cells were found to express Na+/H+-exchange activity only on the basolateral membrane: Na+/H+-exchange activity follows simple saturation kinetics with an apparent K mfor Na+ of approximately 11 mm; it is inhibited in a competitive manner by ethylisopropylamiloride (EIPA). This Na+/H+-exchange activity is inhibited by pharmacological activation of protein kinase A (PKA) as well as of protein kinase C (PKC). Addition of arginine vasopressin (AVP) either at low (subnanomolar) or at high (micromolar) concentrations inhibits Na+/H+-exchange activity; AVP stimulates IP3 production at low concentrations, whereas much higher concentrations are required to stimualte cAMP formation. These findings suggest that in A6 cells (i) Na+/H+-exchange is located in the basolateral membrane and (ii) PKC activation (heralded by IP3 turnover) is likely to be the mediator of AVP action at low AVP concentrations.

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Author information

Author notes

  1. Valeria Casavola & Lorenzo Guerra

    Present address: University of Bari, Institute of Physiology, Via Amendola 165/A, I-70126, Bari, Italy

  2. Stephan J. Reshkin

    Present address: National Institutes of Health, Building 10, Room 1A06, 20892, Bethesda, Maryland

Affiliations

  1. Department of Physiology, University of Zürich, CH-8057, Zürich, Switzerland

    Valeria Casavola, Lorenzo Guerra, Corinna Helmle-Kolb, Stephan J. Reshkin & Heini Murer

Authors
  1. Valeria Casavola
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  2. Lorenzo Guerra
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  3. Corinna Helmle-Kolb
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  4. Stephan J. Reshkin
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  5. Heini Murer
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Additional information

This work was supported by the Swiss National Science Foundation (Grant No. 32-30785.91), the Stiftung für wissenschaftliche Forschung an der Universität Zürich, the Hartmann-Müller Stiftung, the Sandoz-Stiftung, the Roche Research Foundation, and the Geigy Jubiläumsstiftung. Prof. Dr. V. Casavola and Dr. R. Guerra were supported by a research grant, No. 91.02470.CT14 of the Consiglio Nazionale della Ricerche (C.N.R.) We are grateful to Prof. Dr. B.C. Rossier of the Institute of Pharmacology of Lausanne (Switzerland) for the gift of the A6 cells, to H.P. Gaeggeler for the supply of the necessary culture media and to Jutka Forgo for her excellent help in the day-to-day culturing of the A6 cells. The secretarial assistance of D. Rossi is gratefully acknowledged.

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Casavola, V., Guerra, L., Helmle-Kolb, C. et al. Na+/H+-exchange in A6 cells: Polarity and vasopressin regulation. J. Membarin Biol. 130, 105–114 (1992). https://doi.org/10.1007/BF00231889

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  • DOI: https://doi.org/10.1007/BF00231889

Key Words

  • Na+/H+-exchange
  • protein kinase A
  • protein kinase C
  • vasopressin
  • distal tubule
  • tissue culture