Summary
Cultured monolayers of MDCK cells grown upon filter supports display many features ofin vivo epithelia. Previously reported values of transmonolayer resistance of 100 Ωcm−2 (Misfeldt, Hamamoto & Pitelka, 1976; Cereijido, Robbins, Dolan, Rotunno & Sabatini, 1978) indicate a leaky epithelium. This paper describes the properties of a strain of MDCK cells which displays entirely different electrophysiological properties. The results show that (i) the mean transmonolayer resistance is 4.16 kΩ cm−2, (ii) transmonolayer ion transport is of small magnitude since the mean spontaneous open circuit PD is only 2.17 mV basal surface positive and isotopic Na and Cl flux measurements fail to demonstrate a significant net flux, (iii) the action of ouabain, amiloride and ion substitutions are consistent with transmonolayer net Na movement being largely responsible for the spontaneous PD, and (iv) asymmetry in the localization of the Na-K ATPase is evident on the basis of3H-ouabain binding to cell monolayer.
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Abaza, N.A., Leighton, J., Schultz, S.G. 1974. Effects of ouabain on the function and structure of a cell-line (MDCK) derived from canine kidney.In Vitro 10:172
Baker, P.F., Willis, J.F. 1972. Binding of the cardiac glycoside oubain to intact cells.J. Physiol. (London) 244:441
Barker, G., Lamb, J.F., Ogden, P., Simmons, N.L. 1978. The cellular distribution of ouabain binding sites in monolayer cultures of MDCK cells.J. Physiol. (London) 285:46P
Barker, G., Simmons, N.L. 1979. Dog kidney cell monalayers can display properties similar to high-resistance epithelia.J. Physiol. (London) 289:33P
Bindslev, N., Tormey, J., Pietras, R.J., Wright, E.M. 1974. Electrically and osmotically induced changes in permeability and structure of toad urinary bladder.Biochim. Biophys. Acta 332:286
Bisbee, C.A., Machen, T.E., Bern, H.A. 1979. Mouse mammary epithelial cells on floating collagen gels. Transepithelial ion transport and effect of prolactin.Proc. Nat. Acad. Sci. USA 76:540
Boardman, L., Huett, M., Lamb, J.F., Newton, J.P., Polson, J.M. 1974. Evidence for the genetic control of sodium pump density in the Hela cells.J. Physiol (London) 241:677
Boardman, L.J., Lamb, J.F., McCall, D. 1972. Uptake of3H-ouabain and Na pump turnover rates in cells cultured in ouabain.J. Physiol. (London) 225:619
Cabantchik, Z.I., Knauf, P.A., Rothstein, A. 1978. The anion transport system of the red blood cell. The role of membrane protein evaluated by the use of probes.Biochim. Biophys. Acta 515:239
Cereijido, M., Ehrenfeld, J., Meza, I., Martinez-Palomo, A. 1980a. Structural and function membrane polarity in cultured monolayers of MDCK cells.J. Membrane Biol. 52:147
Cereijido, M., Robbins, E.S., Dolan, W.J., Rotunno, C.A., Sabatini, D.D. 1978. Polarized monolayers formed by epithelial cells on a permeable and translucent support.J. Cell Biol. 77:853
Cereijido, M., Stefani, E., Martinez-Palomo, A. 1980b. Occluding junctions in a cultured epithelium: Structural and functional hereogeneity.J. Membrane Biol. 53:19
Cuthbert, A.W., Shum, W.K. 1974. Binding of amiloride to sodium channels in frog skin.Mol. Pharmacol. 10:880
Degnan, K.J., Karnaky, K.J., Zadunaisky, J.A. 1977. Active chloride transport in thein vitro opercular skin of a teleost (Fundulus heteroclitus), a gill-like epithelium rich in chloride cells.J. Physiol. (London) 271:155
Ernst, S.A., Mills, J.W. 1977. Basalateral plasma membrane localization of ouabain-sensitive sodium transport sites in the secretory epithelium of the avian salt gland.J. Cell Biol. 75:74
Frömter, E., Gebler, B. 1977. Electrical properties of amphibian urinary bladder epithelia. III. The cell membrane resistances and the effect of amiloride.Pfluegers Arch. 371:99
Gilbert, S.F., Midgeon, B.R. 1977. Renal enzymes in kidney cells selected byD-valine medium.J. Cell. Physiol. 92:161
Goldring, S.R., Dayer, J.M., Dennis, O.O., Ausiello, D.A., Krane, S.M., 1978. A cell strain cultured from porcine kidney increased cyclic AMP upon exposure to calcitonin or vasopressin.Biochem. Biophys. Res. Commun. 83:434
Handler, J.S., Perkins, F.M., Johnson, J.P. 1980. Studies of renal cell function using cell culture techniques.Am. J. Physiol. 238:F1
Handler, J.S., Steele, R.E., Sahib, M., Wade, J.B., Preston, A.J., Lawson, N., Johnson, J.P. 1979. Toad urinary bladder epithelial cells in culture: Maintenance of epithelial structure, sodium transport and response to hormones.Proc. Nat. Acad. Sci. USA 76:4151
Helman, S.I., Miller, D.A. 1973. Edge damage effect on electrical measurements of frog skin.Am. J. Physiol. 225:972
Higgins, J.T., Gebler, B., Frömter, E. 1975. Electrical properties of urinary amphibian bladder epithelium. 1. Inverse relationship between potential difference and resistance in tightly mounted preparations.Pfluegers. Arch. 358:41
Hossler, F.E., Sarras, M.P., Barnett, R.J. 1978. Ouabain binding during plasma membrane biogenesis in duck salt gland.J. Cell Sci. 31:179
Hossler, F.E., Sarras, M.P., Allen, E. 1978. Ultrastructural, cyto and biochemical observations during turnover of plasma membrane in duck salt gland.Cell Tiss. Res. 188:299
Ludens, J.H., Vaughn, D.A., Mawe, R.C., Fanestil, D.D. 1978. Specific binding of deoxycorticosterone by canine didney cells in culture.J. Steroid Biochem. 9:17
Misfeldt, D.S., Hamamoto, S.T., Pitelka, D.R. 1976. Transepithelial transport in cell culture.Proc. Nat. Acad. Sci. USA 73:1212
Mullin, J.M., Weibel, J., Diamond, K., Kleinzeller, A., 1979. Cell density and the capacity of an established pig renal epithelial cell-line to actively transport sugar.J. Cell Biol. 83:291a
Naftalin, R.J., Simmons, N.L. 1979. The effects of theophylline and choleragen on sodium and chloride ion movements within isolated rabbit ileum.J. Physiol. (London) 290:331
Rabito, C.A., Tchao, R. 1980.3H-ouabain binding during the monolayer organization and cell-cycle in MDCK cells.Am. J. Physiol. 238:C43
Rindler, M.J., Chuman, L.M., Shaffer, L., Saier, M.H., Jr. 1979. Retention of differentiated properties in an established dog kidney epithelial cell line (MDCK).J. Cell Biol. 81:635
Schifferdecker, E., Frömter, E. 1978. The A.C. impedance ofNecturus gallbladder epithelium.Pfluegers Arch. 377:125
Silva, P., Stoff, J., Field, M., Fine, L. 1977. Mechanism of active chloride secretion by shark rectal gland: Role of Na-K ATPase in chloride transport.Am. J. Physiol. 233:F298
Simmons, N.L., Naftalin, R.J. 1976. Bidirectional Na ion movements via the paracellular and transcellular routes across short-circuited rabbit ileum.Biochim. Biophys. Acta 448:426
Snedecor, G.W., Cochran, W.G. 1968. Experimental sampling from a normal population.In: Statistical Methods (6th Ed.) p. 86. Iowa State University Press, Ames
Wedner, H.J., Diamond, J.M. 1969. Contributions of unstirredlayer effects to apparent electrokinetic phenomena in gall-bladder.J. Membrane Biol. 1:92
Welling, D.J., Welling, L.W. 1979. Cell shape as an indicator of volume reabsorption in proximal nephron.Fed. Proc. 38:121
Welling, D.J., Welling, L.W., Hill, J.J. 1978. Phenomenological model relating cell shape to water reabsorption in proximal nephron.Am. J. Physiol. 234:F308
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Simmons, N.L. Ion transport in ‘tight’ epithelial monolayers of MDCK cells. J. Membrain Biol. 59, 105–114 (1981). https://doi.org/10.1007/BF01875708
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DOI: https://doi.org/10.1007/BF01875708