Skip to main content
SpringerLink
Log in

Effect of anisotonic media on volume, ion and amino-acid content and membrane potential of kidney cells (MDCK) in culture

  • Published:
The Journal of Membrane Biology Aims and scope Submit manuscript

Summary

Effects of anisotonic media on a monolayer of confluent kidney cells in culture (MDCK) were studied by measuring: cell thickness and cross-section changes, ion and amino-acid content and membrane potential. The volume was also determined with cells in suspension. When cells in a monolayer were incubated in hypotonic media, the lateral and the apical membranes were rapidly stretched. Afterwards the lateral membranes returned to their initial state while the apical membranes remained stretched. This partial regulatory volume decrease (RVD) was verified with cells in suspension. RVD was accompanied by a loss of K+, Cl− and amino acids, but there was no loss of inorganic phosphate. Also a transient hyperpolarization of the membrane potential was observed, suggesting an increase of the K+ conductance during RVD. Upon restoring the isotonic medium, a regulatory volume increase (RVI) was observed accompanied by a rapid Na+ and Cl− increase and followed by a slow recovery of the initial K+ and Na+ content while amino acids remained at their reduced content. A transient depolarization of the membrane potential was measured during this RVI, suggesting that Na+ and Cl− conductance could have increased. In hypertonic media, only a small and slow RVI was observed accompanied by an increase in K+ and Cl− content but without any change of membrane potential. Quinine partly inhibited RVD in hypotonic media with cells in a monolayer while inhibiting RVD completely with cells in suspension. Incubation during four hours in a Ca2+ free medium had no effect on RVD. Furosemide and amiloride had no effect on RVD and RVI. Volume regulation, RVD or RVI, was not affected by replacing Cl− by nitrate. When cells in a monolayer were incubated in a hypotonic K2SO4 medium, no RVD was observed. From these results, it seems that MDCK cells in a confluent monolayer regulate their volume by activating specific ion and amino-acid transport pathways. Selective K+ and Na+ conductances are activated during RVD and RVI, while the activated anion conductance has a low selectivity. The controlling mechanism might not be the free intracellular Ca2+ concentration.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Aiton, J.F., Brown, C.D.A., Ogden, P., Simmons, N.L. 1982. K+ transport in “tight” epithelial monolayers of MDCK cells.J. Membrane Biol. 65:99–109

    Google Scholar 

  • Brown, C.D.A., Simmons, N.L. 1982. K+ transport in “tight” epithelial monolayers of MDCK cells. Evidence for a calcium-activated K+ channel.Biochim. Biophys. Acta 690:95–105

    PubMed  Google Scholar 

  • Cala, P.M. 1983. Volume regulation by red blood cells. Mechanisms of ion transport.Mol. Physiol. 4:33–52

    Google Scholar 

  • Davis, C.W., Finn, L.A. 1985. Cell volume regulation in frog urinary bladder.Fed. Proc. 44:2520–2525

    PubMed  Google Scholar 

  • DiStefano, A., Wittner, M., Schlatter, E., Lang, H.J., Englert, H., Greger, R. 1985. Diphenylamine-2-carboxylate, a blocker of the Cl− conductive pathway in Cl− transporting epithelia.Pfluegers Arch. 405:S95-S100

    Google Scholar 

  • Eveloff, J., Calamia, J. 1986. Effect of osmolarity on cation fluxes in medullary thick ascending limb cells.Am. J. Physiol. 250:F176-F180

    PubMed  Google Scholar 

  • Eveloff, J.L., Warnock, D.G. 1987. Activation of ion transport systems during cell volume regulation.Am. J. Physiol. 252:F1-F10

    PubMed  Google Scholar 

  • Foskett, J.K., Spring, K.R. 1985. Involvement of calcium and cytoskeleton in gallbladder epithelial cell volume regulation.Am. J. Physiol. 248:C27-C36

    PubMed  Google Scholar 

  • Gagnon, J., Ouimet, D., Nguyen, H., Laprade, R., Le Grimellec, C., Carrière, S., Cardinal, J. 1982. Cell volume regulation in the proximal convoluted tubule.Am. J. Physiol. 243:F408-F415

    PubMed  Google Scholar 

  • Gilles, R. 1979. Intracellular organic osmotic effectors.In: Mechanisms of Osmoregulation in Animals. R. Gilles, editor. pp. 111–154. John Wiley & Sons, New York

    Google Scholar 

  • Grinstein, S., Clarke, C.A., Dupre, A., Rothstein, A. 1982a. Volume-induced increase of anion permeability in human lymphocytes.J. Gen. Physiol. 80:801–823

    PubMed  Google Scholar 

  • Grinstein, S., Clarke, C.A., Rothstein, A. 1983. Activation of Na+/H+ exchange in lymphocytes by osmotically induced volume changes and by cytoplasmic acidification.J. Gen. Physiol. 82:619–638

    PubMed  Google Scholar 

  • Grinstein, S., Dupre, A., Rothstein, A. 1982b. Volume regulation by human lymphocytes. Role of calcium.J. Gen. Physiol. 79:849–868

    PubMed  Google Scholar 

  • Grinstein, S., Rothstein, B., Sarkadi, B., Gelfand, E.W. 1984. Responses of lymphocytes to anisotonic media: Volume-regulating behavior.Am. J. Physiol. 246:C204-C215

    PubMed  Google Scholar 

  • Guggino, S.E., Guggino, W.B., Green, N., Sacktor, B. 1987. Blocking agents of Ca2+-activated K+ channels in cultured medullary thick ascending limb cells.Am. J. Physiol. 252:C128-C137

    PubMed  Google Scholar 

  • Guggino, W.B., Oberleithner, H., Giebisch, G. 1985. Relationship between cell volume and ion transport in the early distal tubule of theAmphiuma kidney.J. Gen. Physiol. 86:31–58

    PubMed  Google Scholar 

  • Herbert, S.C. 1986. Hypertonic cell volume regulation in mouse thick limbs: II. Na+−H+ and Cl−−HCO −3 exchange in basolateral membranes.Am. J. Physiol. 250:C920-C931

    PubMed  Google Scholar 

  • Hoffmann, E. 1977. Control of cell volume.In: Transport of Ions and Water in Animals. B.L. Gupta, R.B. Moreno, J.L. Aschman and B.J. Wall, editors. pp. 285–332, Academic, London

    Google Scholar 

  • Hoffmann, E.K. 1985. Regulatory volume decrease in Ehrlich ascites tumor cells: Role of inorganic ions and amino compounds.Mol. Physiol. 8:167–184

    Google Scholar 

  • Hoffmann, E.K., Lambert, I.H. 1983. Amino acid transport and cell volume regulation in Ehrlich ascites tumor cells.J. Physiol. (London) 338:613–625

    Google Scholar 

  • Hoffmann, E.K., Simonsen, L.O., Lambert, I.H. 1984. Volume-induced increase of K+ and Cl− permeabilities in Ehrlich ascites tumor cells. Role of internal Ca2+.J. Membrane Biol. 78:211–222

    Google Scholar 

  • Kirk, K.L., DiBona, D.R., Schafer, J.A. 1984. Morphologic response of the rabbit cortical collecting tubule to peritubular hypotonicity: Quantitative examination with differential interference contrast microscopy.J. Membrane Biol. 79:53–64

    Google Scholar 

  • Kregenow, F.M. 1981. Osmoregulatory salt transporting mechanisms: Control of cell volume in anisotonic media.Annu. Rev. Physiol. 43:493–505

    PubMed  Google Scholar 

  • Larson, m., Spring, K.R. 1984. Volume regulation byNecturus gallbladder: Basolateral KCl exit.J. Membrane Biol. 81:219–232

    Google Scholar 

  • Lau, K.R., Hudson, R.L., Schultz, S.G. 1984. Cell swelling increases a barium-inhibitable potassium conductance in the basal-lateral membrane ofNecturus small intestine.Proc. Natl. Acad. Sci. USA 81:3591–3594

    PubMed  Google Scholar 

  • Macknight, A.D.C. 1983. Volume regulation in mammalian kidney cells.Mol. Physiol. 4:17–31

    Google Scholar 

  • Paulmichl, M., Gstraunthaler, G., Lang, F. 1985. Electrical properties of Madin-Darby canine kidney cells. Effects of extracellular potassium and bicarbonate.Pfluegers Arch. 405:102–107

    Google Scholar 

  • Richardson, J.C.W., Scalera, V., Simmons, N.L. 1981. Identification of two strains of MDCK cells which resemble separate nephron tubule segments.Biochim. Biophys. Acta 673:26–36

    PubMed  Google Scholar 

  • Rindler, M.J., Saier, M.H. 1981. Evidence for a Na+/H+ antiporter in cultured dog kidney cells.J. Biol. Chem. 256:10820–10824

    PubMed  Google Scholar 

  • Rindler, M.J., Taub, M., Saier, M.H. 1979. Uptake of22Na+ by cultured dog kidney cells (MDCK).J. Biol. Chem. 254:11431–11439

    PubMed  Google Scholar 

  • Roy, G., Sauvé, R. 1983. Stable membrane potentials and mechanical K+ responses activated by internal Ca2+ in HeLa cells.Can. J. Physiol. Pharmacol. 61:144–148

    PubMed  Google Scholar 

  • Simmons, N.L. 1981. The action of ouabain upon chloride secretion in cultured MDCK epithelium.Biochim. Biophys. Acta 646:243–250

    PubMed  Google Scholar 

  • Simmons, N.L. 1984. Epithelial cell volume regulation in hypotonic fluids: Studies using a model tissue culture renal epithelial cell system.Q. J. Exp. Physiol. 69:83–95

    PubMed  Google Scholar 

  • Stefani, E., Cereijido, M. 1983. Electrical properties of cultured epithelioid cells (MDCK).J. Membrane Biol. 73:177–184

    Google Scholar 

  • Strange, K., Spring, K.R. 1987. Cell membrane water permeability of rabbit cortical collecting duct.J. Membrane Biol. 96:27–43

    Google Scholar 

  • Taub, M. 1985. Tissue Culture of Epithelial Cells. Plenum, New York

    Google Scholar 

  • Ussing, H.H. 1985. Volume regulation and basolateral cotransport of sodium, potassium and chloride ions in frog skin epithelium.Pfluegers Arch. 405:S2-S7

    Google Scholar 

  • Welling, P.A., Linshaw, M.A., Sullivan, L.P. 1985. Effect of barium on cell volume regulation in rabbit proximal straight tubules.Am. J. Physiol. 249:F20-F27

    PubMed  Google Scholar 

  • Wong, S.M.E., Chase, H.S. 1986. Role of intracellular calcium in cellular volume regulation.Am. J. Physiol. 250:C841-C852

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Groupe de Recherche en Transport Membranaire, Départments de Physique et de Physiologie, Université de Montréal, Québec, Canada

    G. Roy & R. Sauvé

Authors
  1. G. Roy
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. R. Sauvé
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Roy, G., Sauvé, R. Effect of anisotonic media on volume, ion and amino-acid content and membrane potential of kidney cells (MDCK) in culture. J. Membrain Biol. 100, 83–96 (1987). https://doi.org/10.1007/BF02209143

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02209143

Key Words

Search

Navigation