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. 1984 Nov;3(11):2687–2694. doi: 10.1002/j.1460-2075.1984.tb02194.x

Development of cell surface polarity in the epithelial Madin-Darby canine kidney (MDCK) cell line.

J Balcarova-Ständer, S E Pfeiffer, S D Fuller, K Simons
PMCID: PMC557750  PMID: 6391916

Abstract

The development of surface polarity has been studied in the epithelial Madin-Darby canine kidney (MDCK) cell line by examining two basolateral markers: a monoclonal antibody against a 58-kd protein and [35S]methionine uptake. The surface distribution of these markers was followed after plating the cells on coverslips or nitrocellulose filters. In subconfluent monolayers the apical surface of many cells was stained with the anti-58-kd antibody. Clearing of the apical surface occurred first after confluency had been reached in cells grown on coverslips. Similarly, in cells grown on filters the basolateral 58-kd protein disappeared from the apical surface concomitantly with the development of a measurable electrical resistance over the cell monolayer. The uptake of [35S]methionine was measured from both sides of filter-grown cells and began to polarize early after seeding, reaching a value of greater than 98% basolateral in the fully polarized monolayer. These results emphasize that the development of surface polarity in MDCK cells is a gradual process, and that extensive cell-cell contacts seem to be required for complete surface polarization.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Brandtzaeg P. Conjugates of immunoglobulin G with different fluorochromes. I. Characterization by anionic-exchange chromatography. Scand J Immunol. 1973;2(3):273–290. doi: 10.1111/j.1365-3083.1973.tb02037.x. [DOI] [PubMed] [Google Scholar]
  2. Burke B., Griffiths G., Reggio H., Louvard D., Warren G. A monoclonal antibody against a 135-K Golgi membrane protein. EMBO J. 1982;1(12):1621–1628. doi: 10.1002/j.1460-2075.1982.tb01364.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cereijido M., Robbins E. S., Dolan W. J., Rotunno C. A., Sabatini D. D. Polarized monolayers formed by epithelial cells on a permeable and translucent support. J Cell Biol. 1978 Jun;77(3):853–880. doi: 10.1083/jcb.77.3.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Colas B., Maroux S. Simultaneous isolation of brush border and basolateral membrane from rabbit enterocytes. Presence of brush border hydrolases in the basolateral membrane of rabbit enterocytes. Biochim Biophys Acta. 1980 Aug 4;600(2):406–420. doi: 10.1016/0005-2736(80)90444-7. [DOI] [PubMed] [Google Scholar]
  5. Dobberstein B., Garoff H., Warren G., Robinson P. J. Cell-free synthesis and membrane insertion of mouse H-2Dd histocompatibility antigen and beta 2-microglobulin. Cell. 1979 Aug;17(4):759–769. doi: 10.1016/0092-8674(79)90316-7. [DOI] [PubMed] [Google Scholar]
  6. Herzlinger D. A., Ojakian G. K. Studies on the development and maintenance of epithelial cell surface polarity with monoclonal antibodies. J Cell Biol. 1984 May;98(5):1777–1787. doi: 10.1083/jcb.98.5.1777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hoi Sang U., Saier M. H., Jr, Ellisman M. H. Tight junction formation is closely linked to the polar redistribution of intramembranous particles in aggregating MDCK epithelia. Exp Cell Res. 1979 Sep;122(2):384–391. doi: 10.1016/0014-4827(79)90315-x. [DOI] [PubMed] [Google Scholar]
  8. Hynes R. O. Alteration of cell-surface proteins by viral transformation and by proteolysis. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3170–3174. doi: 10.1073/pnas.70.11.3170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Louvard D. Apical membrane aminopeptidase appears at site of cell-cell contact in cultured kidney epithelial cells. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4132–4136. doi: 10.1073/pnas.77.7.4132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Matlin K. S., Simons K. Reduced temperature prevents transfer of a membrane glycoprotein to the cell surface but does not prevent terminal glycosylation. Cell. 1983 Aug;34(1):233–243. doi: 10.1016/0092-8674(83)90154-x. [DOI] [PubMed] [Google Scholar]
  13. Matlin K., Bainton D. F., Pesonen M., Louvard D., Genty N., Simons K. Transepithelial transport of a viral membrane glycoprotein implanted into the apical plasma membrane of Madin-Darby canine kidney cells. I. Morphological evidence. J Cell Biol. 1983 Sep;97(3):627–637. doi: 10.1083/jcb.97.3.627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Meier P. J., Sztul E. S., Reuben A., Boyer J. L. Structural and functional polarity of canalicular and basolateral plasma membrane vesicles isolated in high yield from rat liver. J Cell Biol. 1984 Mar;98(3):991–1000. doi: 10.1083/jcb.98.3.991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Misfeldt D. S., Hamamoto S. T., Pitelka D. R. Transepithelial transport in cell culture. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1212–1216. doi: 10.1073/pnas.73.4.1212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Perkins F. M., Handler J. S. Transport properties of toad kidney epithelia in culture. Am J Physiol. 1981 Sep;241(3):C154–C159. doi: 10.1152/ajpcell.1981.241.3.C154. [DOI] [PubMed] [Google Scholar]
  17. Pesonen M., Simons K. Transepithelial transport of a viral membrane glycoprotein implanted into the apical plasma membrane of Madin-Darby canine kidney cells. II. Immunological quantitation. J Cell Biol. 1983 Sep;97(3):638–643. doi: 10.1083/jcb.97.3.638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pisam M., Ripoche P. Redistribution of surface macromolecules in dissociated epithelial cells. J Cell Biol. 1976 Dec;71(3):907–920. doi: 10.1083/jcb.71.3.907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rabito C. A., Karish M. V. Polarized amino acid transport by an epithelial cell line of renal origin (LLC-PK1). The apical systems. J Biol Chem. 1983 Feb 25;258(4):2543–2547. [PubMed] [Google Scholar]
  20. Rabito C. A., Karish M. V. Polarized amino acid transport by an epithelial cell line of renal origin (LLC-PK1). The basolateral systems. J Biol Chem. 1982 Jun 25;257(12):6802–6808. [PubMed] [Google Scholar]
  21. Richardson J. C., Scalera V., Simmons N. L. Identification of two strains of MDCK cells which resemble separate nephron tubule segments. Biochim Biophys Acta. 1981 Feb 18;673(1):26–36. [PubMed] [Google Scholar]
  22. Richardson J. C., Simmons N. L. Demonstration of protein asymmetries in the plasma membrane of cultured renal (MDCK) epithelial cells by lactoperoxidase-mediated iodination. FEBS Lett. 1979 Sep 15;105(2):201–204. doi: 10.1016/0014-5793(79)80611-0. [DOI] [PubMed] [Google Scholar]
  23. Rodriguez-Boulan E., Paskiet K. T., Sabatini D. D. Assembly of enveloped viruses in Madin-Darby canine kidney cells: polarized budding from single attached cells and from clusters of cells in suspension. J Cell Biol. 1983 Mar;96(3):866–874. doi: 10.1083/jcb.96.3.866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shulman M., Wilde C. D., Köhler G. A better cell line for making hybridomas secreting specific antibodies. Nature. 1978 Nov 16;276(5685):269–270. doi: 10.1038/276269a0. [DOI] [PubMed] [Google Scholar]
  25. Simons K., Garoff H. The budding mechanisms of enveloped animal viruses. J Gen Virol. 1980 Sep;50(1):1–21. doi: 10.1099/0022-1317-50-1-1. [DOI] [PubMed] [Google Scholar]
  26. U H. S., Evans-Layng M. Polar redistribution of Na+K+ATPase in aggregating MDCK cells. Exp Cell Res. 1983 Jun;146(1):192–198. doi: 10.1016/0014-4827(83)90337-3. [DOI] [PubMed] [Google Scholar]
  27. Ziomek C. A., Schulman S., Edidin M. Redistribution of membrane proteins in isolated mouse intestinal epithelial cells. J Cell Biol. 1980 Sep;86(3):849–857. doi: 10.1083/jcb.86.3.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. van Meer G., Simons K. Viruses budding from either the apical or the basolateral plasma membrane domain of MDCK cells have unique phospholipid compositions. EMBO J. 1982;1(7):847–852. doi: 10.1002/j.1460-2075.1982.tb01258.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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