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PMID: 567227 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Polarized monolayers formed by epithelial cells on a permeable and translucent support.

The Journal of cell biology ·Vol. 77 ·No. 3 ·1978-06-00 ·Pages 853-80

Cereijido M, Robbins ES, Dolan WJ, Rotunno CA, Sabatini DD

Abstract

An epithelial cell line (MDCK) was used to prepare monolayers which, in vitro, develop properties of transporting epithelia. Monolayers were formed by plating cells at high densities (10(6) cells/cm2) on collagen-coated nylon cloth disks to saturate the area available for attachment, thus avoiding the need for cell division. An electrical resistance developed within 4-6 h after plating and achieved a steady-state value of 104 +/- 1.8 omega-cm2 after 24 h. Mature monolayers were morphologically and functionally polarized. They contained junctional complexes composed of desmosomes and tight junctions with properties similar to those of "leaky" epithelia. Monolayers were capable of maintaining a spontaneous electrical potential sensitive to amiloride, produced a net water flux from the apical to basal side, and discriminated between Na+ and Cl- ions. The MDCK permeability barrier behaves as a "thin" membrane with negatively charged sites. It has: (a) a linear conductance/concentration relationship; (b) an asymmetric instantaneous current/voltage relationship; (c) a reduced ability to discriminate between Na+ and Cl- caused by lowering the pH; and (d) a characteristic pattern of ionic selectivity which suggests that the negatively charged sites are highly hydrates and of medium field strength. Measurements of Na+ permeability of electrical and tracer methods ruled out exchange diffusion as a mechanism for ion permeation and the lack of current saturation in the I/deltapsi curves does not support the involvement of carriers. The discrimination between Na+ and Cl- was severely but reversibly decreased at low pH, suggesting that Na+-specific channels which exclude Cl- contain acidic groups dissociated at neutral pH. Bound Ca++ ions are involved in maintaining the integrity of the junctions in MDCK monolayers as was shown by a reversible drop of resistance and opening of the junctions in Ca++-free medium containing EGTA. Several other epithelial cell lines are capable of developing a significant resistance under the conditions used to obtain MDCK monolayers.

MeSH Terms
Animals Cell Line Cell Membrane Permeability Chlorides/metabolism Culture Techniques/instrumentation Dogs Electric Conductivity Epithelial Cells Epithelium/physiology,ultrastructure Intercellular Junctions/ultrastructure Membrane Potentials Sodium/metabolism Water/metabolism
Chemicals
Chlorides Water Sodium
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Cereijido M
Robbins E S
Dolan W J
Rotunno C A
Sabatini D D
References (63)
63 references, click to expand
  1. Cation permeation mechanisms and cation selectivity in "tight junctions" of gallbladder epithelium.
    Membranes. 1975;3:383-497 PMID: 587
  2. Intercellular junctions of oral epithelium. I. Studies with freeze-fracture and tracing methods of normal rat keratinized oral epithelium.
    J Ultrastruct Res. 1976 Jul;56(1):121-36 PMID: 59813
  3. The desmosome: fine structural studies with freeze-fracture replication and tannic acid staining of sectioned epidermis.
    Cell Tissue Res. 1976 Sep 20;172(3):309-23 PMID: 62618
  4. Structure of tight junctions in epithelia with different permeability.
    Proc Natl Acad Sci U S A. 1975 Nov;72(11):4487-91 PMID: 812096
  5. Extracellular calcium and the organization of tight junctions in pancreatic acinar cells.
    Exp Cell Res. 1976 Apr;99(1):178-83 PMID: 816663
  6. Velocity distributions of the streaming protoplasm in Nitella flexilis.
    Biophys J. 1977 Mar;17(3):229-41 PMID: 843582
  7. Lack of correlation between tight junction morphology and permeability properties in developing choroid plexus.
    Nature. 1976 Nov 18;264(5583):293-4 PMID: 1004553
  8. Establishment of tight junctions between epithelial cells.
    Proc Natl Acad Sci U S A. 1975 Jul;72(7):2711-3 PMID: 1058486
  9. Transepithelial transport in cell culture.
    Proc Natl Acad Sci U S A. 1976 Apr;73(4):1212-6 PMID: 1063404
  10. The role of the lateral intercellular spaces in the control of ion permeation across the rabbit gall bladder.
    Pflugers Arch. 1975 Jul 9;358(1):27-40 PMID: 1172236
  11. Vitamin-A-induced mucous metaplasia. An in vitro system for modulating tight and gap junction differentiation.
    J Cell Biol. 1976 Feb;68(2):173-88 PMID: 1245547
  12. A fine structural analysis of intercellular junctions in the mouse liver.
    J Cell Biol. 1970 May;45(2):272-90 PMID: 4105112
  13. A cell line derived from normal dog kidney (MDCK) exhibiting qualities of papillary adenocarcinoma and of renal tubular epithelium.
    Cancer. 1970 Nov;26(5):1022-8 PMID: 4248968
  14. The study of cellular "microexudates" by ellipsometry and their relationship to the cell coat.
    Exp Cell Res. 1973 Apr;78(2):303-13 PMID: 4349014
  15. Preparation and characterization of an immuno-electron microscope tracer consisting of a heme-octapeptide coupled to fab.
    J Exp Med. 1974 Jan 1;139(1):208-23 PMID: 4357686
  16. Electrical properties of the medullary collecting ducts of the golden hamster kidney. II. The transepithelial resistance.
    Pflugers Arch. 1974;351(2):113-31 PMID: 4473141
  17. Route of passive ion permeation in epithelia.
    Nat New Biol. 1972 Jan 5;235(53):9-13 PMID: 4502409
  18. Mechanisms of ion transport across the choroid plexus.
    J Physiol. 1972 Oct;226(2):545-71 PMID: 4538945
  19. Discrimination of monovalent inorganic cations by "tight" junctions of gallbladder epithelium.
    J Membr Biol. 1974;15(3):277-318 PMID: 4546135
  20. Role of diffusion boundary layer in contact inhibition of growth.
    Nature. 1973 Nov 23;246(5430):200-3 PMID: 4586793
  21. The upper cell surface: its inability to support active cell movement in culture.
    J Cell Biol. 1974 Jul;62(1):198-214 PMID: 4600884
  22. The distribution of lanthanum in tight junctions of the kidney tubule.
    Pflugers Arch. 1973 Oct 22;343(3):267-72 PMID: 4796785
  23. A freeze-etch study of the tight junctions of the rat kidney tubules.
    Lab Invest. 1974 Mar;30(3):286-91 PMID: 4817823
  24. The sodium-transporting compartment of the epithelium of frog skin.
    J Physiol. 1974 Mar;237(3):555-71 PMID: 4825475
  25. Evidence of a paracellular pathway for ion flow in the kidney proximal tubule. Electromicroscopic demonstration of lanthanum precipitate in the tight junction.
    Pflugers Arch. 1971;330(4):302-9 PMID: 5004222
  26. Sodium flux in Necturus proximal tubule under voltage clamp.
    J Gen Physiol. 1972 Aug;60(2):181-201 PMID: 5049078
  27. Ionic conductances of extracellular shunt pathway in rabbit ileum. Influence of shunt on transmural sodium transport and electrical potential differences.
    J Gen Physiol. 1972 Mar;59(3):318-46 PMID: 5058963
  28. Some electrical properties of distal tubular epithelium in the rat.
    Am J Physiol. 1972 Oct;223(4):797-808 PMID: 5075156
  29. Effect of vasopressin on electrical resistance of renal cortical collecting tubules.
    Am J Physiol. 1971 Jun;220(6):1825-32 PMID: 5087832
  30. Electrophysiology of proximal and distal tubules in the autoperfused dog kidney.
    Am J Physiol. 1971 Oct;221(4):1084-96 PMID: 5111249
  31. Membrane structure and ion permeation. Study of ion exchange membrane structure and function is relevant to analysis of biological ion permeation.
    Science. 1967 Feb 24;155(3765):965-74 PMID: 5334938
  32. Changes in sodium pool and kinetics of sodium transport in frog skin produced by amiloride.
    Br J Pharmacol. 1970 May;39(1):99-109 PMID: 5420148
  33. The hyperpolarizing region of the current-voltage curve in frog skin.
    Biophys J. 1970 Apr;10(4):323-44 PMID: 5436882
  34. Ion transport across the frog posterior choroid plexus.
    Brain Res. 1970 Oct 13;23(2):302-4 PMID: 5476777
  35. The ion permeability induced in thin lipid membranes by the polyene antibiotics nystatin and amphotericin B.
    J Gen Physiol. 1970 Jul;56(1):100-24 PMID: 5514157
  36. Effect of amiloride on sodium transport in frog skin. II. Sodium transport pool and unidirectional fluxes.
    Pflugers Arch. 1970;321(2):91-101 PMID: 5529540
  37. Edge damage effect in in vitro frog skin preparations.
    Am J Physiol. 1968 Apr;214(4):719-24 PMID: 5642931
  38. Amiloride: a potent inhibitor of sodium transport across the toad bladder.
    J Physiol. 1968 Mar;195(2):317-30 PMID: 5647323
  39. Effects of pH and polyvalent cations on the selective permeability of gall-bladder epithelium to monovalent ions.
    Biochim Biophys Acta. 1968 Aug;163(1):57-74 PMID: 5666778
  40. Effect of external and internal pH changes on K and Cl conductances in the muscle fiber membrane of a giant barnacle.
    J Gen Physiol. 1968 Nov;52(5):773-92 PMID: 5688083
  41. Induced pacemaker activity on toad skin.
    J Gen Physiol. 1968 Jun;51(6):785-801 PMID: 5692095
  42. Secretory activity and oncogenicity of a cell line (MDCK) derived from canine kidney.
    Science. 1969 Jan 31;163(3866):472-3 PMID: 5762397
  43. Electroosmosis in membranes: effects of unstirred layers and transport numbers. II. Experimental.
    Biophys J. 1969 May;9(5):729-57 PMID: 5786318
  44. The initiation of cell division in a contact-inhibited mammalian cell line.
    J Cell Physiol. 1965 Dec;66(3):325-33 PMID: 5884360
  45. Studies on ion accumulation in muscle cells.
    J Gen Physiol. 1966 Mar;49(4):819-43 PMID: 5943617
  46. The steady-state properties of an ion exchange membrane with mobile sites.
    Biophys J. 1966 May;6(3):227-46 PMID: 5962278
  47. Interpretation of hexose-dependent electrical potential differences in small intestine.
    Nature. 1967 Apr 29;214(5087):509-10 PMID: 6032886
  48. Hexagonal array of subunits in intercellular junctions of the mouse heart and liver.
    J Cell Biol. 1967 Jun;33(3):C7-C12 PMID: 6036535
  49. The pH sensitivity of the chloride conductance of frog skeletal muscle.
    J Physiol. 1967 Apr;189(3):403-25 PMID: 6040154
  50. Factors governing the transepithelial potential difference across the proximal tubule of the rat kidney.
    J Clin Invest. 1974 Feb;53(2):454-64 PMID: 11344559
  51. The nature of the frog skin potential.
    Acta Physiol Scand. 1958 Jun 2;42(3-4):298-308 PMID: 13544986
  52. The reabsorptive function of the gall-bladder.
    J Physiol. 1962 May;161:442-73 PMID: 13886112
  53. Cation selective glass electrodes and their mode of operation.
    Biophys J. 1962 Mar;2(2 Pt 2):259-323 PMID: 13889686
  54. Junctional complexes in various epithelia.
    J Cell Biol. 1963 May;17:375-412 PMID: 13944428
  55. Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation.
    J Cell Biol. 1963 Apr;17:19-58 PMID: 13975866
  56. The interaction between active sodium transport and active sugar transport in the isolated rabbit ileum.
    Biochim Biophys Acta. 1963 May 14;71:503-5 PMID: 13987238
  57. CHEMICAL CONCENTRATION GRADIENTS AND ELECTRICAL PROPERTIES OF GASTRIC MUCOSA.
    Am J Physiol. 1964 Apr;206:769-82 PMID: 14166171
  58. NATURE OF SHUNT PATH AND ACTIVE SODIUM TRANSPORT PATH THROUGH FROG SKIN EPITHELIUM.
    Acta Physiol Scand. 1964 Aug;61:484-504 PMID: 14209264
  59. [AN AUTOMATIC DEVICE FOR MEASURING AND RECORDING THE NET FLOW OF WATER THROUGH THE SKIN AND BLADDER OF AMPHIBIA].
    Biochim Biophys Acta. 1964 Sep 25;88:442-4 PMID: 14253227
  60. EFFECT OF ALKALI METAL CATIONS ON THE POTENTIAL ACROSS TOAD AND BULLFROG URINARY BLADDER.
    J Gen Physiol. 1965 Jan;48:527-40 PMID: 14284782
  61. INTRACELLULAR ELECTRICAL POTENTIALS IN FROG SKIN.
    J Gen Physiol. 1965 Mar;48:543-57 PMID: 14324974
  62. Some aspects of the application of tracers in permeability studies.
    Adv Enzymol Relat Subj Biochem. 1952;13:21-65 PMID: 14943664
  63. Coupling and selectivity of sodium and potassium transport in squid giant axons.
    J Gen Physiol. 1968 May 1;51(5):152-61 PMID: 19873583
Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.77.3.853
Published
1978-06-00
Pages
853-80
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2110138
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.77.3.853 PMC Full Text View journal details
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