The Journal of Membrane Biology

, Volume 65, Issue 1–2, pp 99–109 | Cite as

K+ transport in ‘Tight’ epithelial monolayers of MDCK cells

  • J. F. Aiton
  • C. D. A. Brown
  • P. Ogden
  • N. L. Simmons
Articles

Summary

Bidirectional transepithelial K+ flux measurements across ‘high-resistance’ epithelial monolayers of MDCK cells grown upon millipore filters show no significant net K+ flux.

Measurements of influx and efflux across the basal-lateral and apical cell membranes demonstrate that the apical membranes are effectively impermeable to K+.

K+ influx across the basal-lateral cell membranes consists of an ouabain-sensitive component, an ouabain-insensitive component, an ouabain-insensitive but furosemide-sensitive component, and an ouabain-and furosemide-insensitive component.

The action of furosemide upon K+ influx is independent of (Na+−K+)-pump inhibition. The furosemide-sensitive component is markedly dependent upon the medium K+, Na+ and Cl content. Acetate and nitrate are ineffective substitutes for Cl, whereas Br is partially effective. Partial Cl replacement by NO3 gives a roughly linear increase in the furosemide-sensitive component. Na+ replacement by choline abolishes the furosemide-sensitive component, whereas Li+ is a partially effective replacement. Partial Na+ replacement with choline gives an apparent affinity of ∼7mm Na, whereas variation of the external K+ content gives an affinity of the furosemide-sensitive component of 1.0mm.

Furosemide inhibition is of high affinity (K1/2=3 μm). Piretanide, ethacrynic acid, and phloretin inhibit the same component of passive K+ influx as furosemide; amiloride, 4,-aminopyridine, and 2,4,6-triaminopyrimidine partially so. SITS was ineffective.

Externally applied furosemide and Cl replacement by NO 3 inhibit K+ efflux across the basal-lateral membranes indicating that the furosemide-sensitive component consists primarily of K∶K exchange.

Key words

K+ fluxes MDCK ouabain furosemide cultured epithelium Na++K++Cl cotransport 

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Copyright information

© Springer-Verlag New York Inc 1982

Authors and Affiliations

  • J. F. Aiton
    • 1
  • C. D. A. Brown
    • 1
  • P. Ogden
    • 1
  • N. L. Simmons
    • 1
  1. 1.The Department of PhysiologyUniversity of St.AndrewsSt.AndrewsScotland

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