Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1989 Mar 1;108(3):903–919. doi: 10.1083/jcb.108.3.903

Type I collagen gel induces Madin-Darby canine kidney cells to become fusiform in shape and lose apical-basal polarity

PMCID: PMC2115371  PMID: 2537838

Abstract

In the embryo, epithelia give rise to mesenchyme at specific times and places. Recently, it has been reported (Greenburg, G., and E. D. Hay. 1986. Dev. Biol. 115:363-379; Greenberg, G., and E. D. Hay. 1988. Development (Camb.). 102:605-622) that definitive epithelia can give rise to fibroblast-like cells when suspended within type I collagen gels. We wanted to know whether Madin-Darby canine kidney (MDCK) cells, an epithelial line, can form mesenchyme under similar conditions. Small explants of MDCK cells on basement membrane were suspended within or placed on top of extracellular matrix gels. MDCK cells on basement membrane gel are tall, columnar in shape, and ultrastructurally resemble epithelia transporting fluid and ions. MDCK explants cultured on type I collagen gel give rise to isolated fusiform-shaped cells that migrate over the gel surface. The fusiform cells extend pseudopodia and filopodia, lose cell membrane specializations, and develop an actin cortex around the entire cell. Unlike true mesenchymal cells, which express vimentin and type I collagen, fusiform cells produce both keratin and vimentin, continue to express laminin, and do not turn on type I collagen. Fusiform cells are not apically-basally polarized, but show mesenchymal cell polarity. Influenza hemagglutinin and virus budding localize to the front end or entire cell surface. Na,K-ATPase occurs intracellularly and also symmetrically distributes on the cell surface. Fodrin becomes diffusely distributed along the plasma membrane, ZO-1 cannot be detected, and desmoplakins distribute randomly in the cytoplasm. The loss of epithelial polarity and acquisition of mesenchymal cell polarity and shape by fusiform MDCK cells on type I collagen gel was previously unsuspected. The phenomenon may offer new opportunities for studying cytoplasmic and nuclear mechanisms regulating cell shape and polarity.

Full Text

The Full Text of this article is available as a PDF (8.5 MB).

Selected References

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

  1. Abercrombie M., Heaysman J. E., Pegrum S. M. The locomotion of fibroblasts in culture. 3. Movements of particles on the dorsal surface of the leading lamella. Exp Cell Res. 1970 Oct;62(2):389–398. doi: 10.1016/0014-4827(70)90570-7. [DOI] [PubMed] [Google Scholar]
  2. Bard J. B., Hay E. D. The behavior of fibroblasts from the developing avian cornea. Morphology and movement in situ and in vitro. J Cell Biol. 1975 Nov;67(2PT1):400–418. doi: 10.1083/jcb.67.2.400. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ben-Ze'ev A. Differential control of cytokeratins and vimentin synthesis by cell-cell contact and cell spreading in cultured epithelial cells. J Cell Biol. 1984 Oct;99(4 Pt 1):1424–1433. doi: 10.1083/jcb.99.4.1424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bergmann J. E., Kupfer A., Singer S. J. Membrane insertion at the leading edge of motile fibroblasts. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1367–1371. doi: 10.1073/pnas.80.5.1367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Caplan M. J., Stow J. L., Newman A. P., Madri J., Anderson H. C., Farquhar M. G., Palade G. E., Jamieson J. D. Dependence on pH of polarized sorting of secreted proteins. Nature. 1987 Oct 15;329(6140):632–635. doi: 10.1038/329632a0. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Dodson J. W., Hay E. D. Secretion of collagen by corneal epithelium. II. Effect of the underlying substratum on secretion and polymerization of epithelial products. J Exp Zool. 1974 Jul;189(1):51–72. doi: 10.1002/jez.1401890106. [DOI] [PubMed] [Google Scholar]
  8. Elsdale T., Bard J. Collagen substrata for studies on cell behavior. J Cell Biol. 1972 Sep;54(3):626–637. doi: 10.1083/jcb.54.3.626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fitchett J. E., Hay E. D. Medial edge epithelium transforms to mesenchyme after embryonic palatal shelves fuse. Dev Biol. 1989 Feb;131(2):455–474. doi: 10.1016/s0012-1606(89)80017-x. [DOI] [PubMed] [Google Scholar]
  10. Franke W. W., Grund C., Kuhn C., Jackson B. W., Illmensee K. Formation of cytoskeletal elements during mouse embryogenesis. III. Primary mesenchymal cells and the first appearance of vimentin filaments. Differentiation. 1982;23(1):43–59. doi: 10.1111/j.1432-0436.1982.tb01266.x. [DOI] [PubMed] [Google Scholar]
  11. Franke W. W., Schmid E., Breitkreutz D., Lüder M., Boukamp P., Fusenig N. E., Osborn M., Weber K. Simultaneous expression of two different types of intermediate sized filaments in mouse keratinocytes proliferating in vitro. Differentiation. 1979;14(1-2):35–50. doi: 10.1111/j.1432-0436.1979.tb01010.x. [DOI] [PubMed] [Google Scholar]
  12. Fuller S., von Bonsdorff C. H., Simons K. Vesicular stomatitis virus infects and matures only through the basolateral surface of the polarized epithelial cell line, MDCK. Cell. 1984 Aug;38(1):65–77. doi: 10.1016/0092-8674(84)90527-0. [DOI] [PubMed] [Google Scholar]
  13. Garcia-Perez A., Smith W. L. Use of monoclonal antibodies to isolate cortical collecting tubule cells: AVP induces PGE release. Am J Physiol. 1983 Mar;244(3):C211–C220. doi: 10.1152/ajpcell.1983.244.3.C211. [DOI] [PubMed] [Google Scholar]
  14. Glenney J. R., Jr, Glenney P. Comparison of spectrin isolated from erythroid and non-erythroid sources. Eur J Biochem. 1984 Nov 2;144(3):529–539. doi: 10.1111/j.1432-1033.1984.tb08498.x. [DOI] [PubMed] [Google Scholar]
  15. Greenburg G., Hay E. D. Cytodifferentiation and tissue phenotype change during transformation of embryonic lens epithelium to mesenchyme-like cells in vitro. Dev Biol. 1986 Jun;115(2):363–379. doi: 10.1016/0012-1606(86)90256-3. [DOI] [PubMed] [Google Scholar]
  16. Greenburg G., Hay E. D. Cytoskeleton and thyroglobulin expression change during transformation of thyroid epithelium to mesenchyme-like cells. Development. 1988 Mar;102(3):605–622. doi: 10.1242/dev.102.3.605. [DOI] [PubMed] [Google Scholar]
  17. Greenburg G., Hay E. D. Epithelia suspended in collagen gels can lose polarity and express characteristics of migrating mesenchymal cells. J Cell Biol. 1982 Oct;95(1):333–339. doi: 10.1083/jcb.95.1.333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Haeuptle M. T., Suard Y. L., Bogenmann E., Reggio H., Racine L., Kraehenbuhl J. P. Effect of cell shape change on the function and differentiation of rabbit mammary cells in culture. J Cell Biol. 1983 May;96(5):1425–1434. doi: 10.1083/jcb.96.5.1425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Harris A. S., Anderson J. P., Yurchenco P. D., Green L. A., Ainger K. J., Morrow J. S. Mechanisms of cytoskeletal regulation: functional and antigenic diversity in human erythrocyte and brain beta spectrin. J Cell Biochem. 1986;30(1):51–69. doi: 10.1002/jcb.240300107. [DOI] [PubMed] [Google Scholar]
  20. Herzlinger D. A., Easton T. G., Ojakian G. K. The MDCK epithelial cell line expresses a cell surface antigen of the kidney distal tubule. J Cell Biol. 1982 May;93(2):269–277. doi: 10.1083/jcb.93.2.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kaye G. I., Wheeler H. O., Whitlock R. T., Lane N. Fluid transport in the rabbit gallbladder. A combined physiological and electron microscopic study. J Cell Biol. 1966 Aug;30(2):237–268. doi: 10.1083/jcb.30.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kupfer A., Kronebusch P. J., Rose J. K., Singer S. J. A critical role for the polarization of membrane recycling in cell motility. Cell Motil Cytoskeleton. 1987;8(2):182–189. doi: 10.1002/cm.970080210. [DOI] [PubMed] [Google Scholar]
  23. Kupfer A., Louvard D., Singer S. J. Polarization of the Golgi apparatus and the microtubule-organizing center in cultured fibroblasts at the edge of an experimental wound. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2603–2607. doi: 10.1073/pnas.79.8.2603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lawrence J. B., Singer R. H. Intracellular localization of messenger RNAs for cytoskeletal proteins. Cell. 1986 May 9;45(3):407–415. doi: 10.1016/0092-8674(86)90326-0. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Matlin K. S. Ammonium chloride slows transport of the influenza virus hemagglutinin but does not cause mis-sorting in a polarized epithelial cell line. J Biol Chem. 1986 Nov 15;261(32):15172–15178. [PubMed] [Google Scholar]
  27. Matlin K. S., Reggio H., Helenius A., Simons K. Infectious entry pathway of influenza virus in a canine kidney cell line. J Cell Biol. 1981 Dec;91(3 Pt 1):601–613. doi: 10.1083/jcb.91.3.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. McAteer J. A., Evan A. P., Gardner K. D. Morphogenetic clonal growth of kidney epithelial cell line MDCK. Anat Rec. 1987 Mar;217(3):229–239. doi: 10.1002/ar.1092170303. [DOI] [PubMed] [Google Scholar]
  30. McDonald J. A., Broekelmann T. J., Matheke M. L., Crouch E., Koo M., Kuhn C., 3rd A monoclonal antibody to the carboxyterminal domain of procollagen type I visualizes collagen-synthesizing fibroblasts. Detection of an altered fibroblast phenotype in lungs of patients with pulmonary fibrosis. J Clin Invest. 1986 Nov;78(5):1237–1244. doi: 10.1172/JCI112707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Mueller H., Franke W. W. Biochemical and immunological characterization of desmoplakins I and II, the major polypeptides of the desmosomal plaque. J Mol Biol. 1983 Feb 5;163(4):647–671. doi: 10.1016/0022-2836(83)90116-x. [DOI] [PubMed] [Google Scholar]
  33. Nelson W. J., Veshnock P. J. Dynamics of membrane-skeleton (fodrin) organization during development of polarity in Madin-Darby canine kidney epithelial cells. J Cell Biol. 1986 Nov;103(5):1751–1765. doi: 10.1083/jcb.103.5.1751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Nelson W. J., Veshnock P. J. Modulation of fodrin (membrane skeleton) stability by cell-cell contact in Madin-Darby canine kidney epithelial cells. J Cell Biol. 1987 Jun;104(6):1527–1537. doi: 10.1083/jcb.104.6.1527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Osborn M., Weber K. Tumor diagnosis by intermediate filament typing: a novel tool for surgical pathology. Lab Invest. 1983 Apr;48(4):372–394. [PubMed] [Google Scholar]
  36. Pasdar M., Nelson W. J. Kinetics of desmosome assembly in Madin-Darby canine kidney epithelial cells: temporal and spatial regulation of desmoplakin organization and stabilization upon cell-cell contact. I. Biochemical analysis. J Cell Biol. 1988 Mar;106(3):677–685. doi: 10.1083/jcb.106.3.677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Pasdar M., Nelson W. J. Kinetics of desmosome assembly in Madin-Darby canine kidney epithelial cells: temporal and spatial regulation of desmoplakin organization and stabilization upon cell-cell contact. II. Morphological analysis. J Cell Biol. 1988 Mar;106(3):687–695. doi: 10.1083/jcb.106.3.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ramaekers F. C., Osborn M., Schimid E., Weber K., Bloemendal H., Franke W. W. Identification of the cytoskeletal proteins in lens-forming cells, a special epitheloid cell type. Exp Cell Res. 1980 Jun;127(2):309–327. doi: 10.1016/0014-4827(80)90437-1. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Rindler M. J., Chuman L. M., Shaffer L., Saier M. H., Jr Retention of differentiated properties in an established dog kidney epithelial cell line (MDCK). J Cell Biol. 1979 Jun;81(3):635–648. doi: 10.1083/jcb.81.3.635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Rindler M. J., Ivanov I. E., Sabatini D. D. Microtubule-acting drugs lead to the nonpolarized delivery of the influenza hemagglutinin to the cell surface of polarized Madin-Darby canine kidney cells. J Cell Biol. 1987 Feb;104(2):231–241. doi: 10.1083/jcb.104.2.231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Rodriguez Boulan E., Sabatini D. D. Asymmetric budding of viruses in epithelial monlayers: a model system for study of epithelial polarity. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5071–5075. doi: 10.1073/pnas.75.10.5071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. 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]
  44. Rogalski A. A., Bergmann J. E., Singer S. J. Effect of microtubule assembly status on the intracellular processing and surface expression of an integral protein of the plasma membrane. J Cell Biol. 1984 Sep;99(3):1101–1109. doi: 10.1083/jcb.99.3.1101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Salas P. J., Misek D. E., Vega-Salas D. E., Gundersen D., Cereijido M., Rodriguez-Boulan E. Microtubules and actin filaments are not critically involved in the biogenesis of epithelial cell surface polarity. J Cell Biol. 1986 May;102(5):1853–1867. doi: 10.1083/jcb.102.5.1853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Schmid E., Tapscott S., Bennett G. S., Croop J., Fellini S. A., Holtzer H., Franke W. W. Differential location of different types of intermediate-sized filaments in various tissues of the chicken embryo. Differentiation. 1979;15(1):27–40. doi: 10.1111/j.1432-0436.1979.tb01031.x. [DOI] [PubMed] [Google Scholar]
  47. Simons K., Fuller S. D. Cell surface polarity in epithelia. Annu Rev Cell Biol. 1985;1:243–288. doi: 10.1146/annurev.cb.01.110185.001331. [DOI] [PubMed] [Google Scholar]
  48. Smith Z. D., Caplan M. J., Jamieson J. D. Immunocytochemical localization of plasmalemmal proteins in semi-thin sections of epithelial monolayers. J Histochem Cytochem. 1988 Mar;36(3):311–316. doi: 10.1177/36.3.2449492. [DOI] [PubMed] [Google Scholar]
  49. Stevenson B. R., Siliciano J. D., Mooseker M. S., Goodenough D. A. Identification of ZO-1: a high molecular weight polypeptide associated with the tight junction (zonula occludens) in a variety of epithelia. J Cell Biol. 1986 Sep;103(3):755–766. doi: 10.1083/jcb.103.3.755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Tomasek J. J., Hay E. D. Analysis of the role of microfilaments and microtubules in acquisition of bipolarity and elongation of fibroblasts in hydrated collagen gels. J Cell Biol. 1984 Aug;99(2):536–549. doi: 10.1083/jcb.99.2.536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Tomasek J. J., Hay E. D., Fujiwara K. Collagen modulates cell shape and cytoskeleton of embryonic corneal and fibroma fibroblasts: distribution of actin, alpha-actinin, and myosin. Dev Biol. 1982 Jul;92(1):107–122. doi: 10.1016/0012-1606(82)90155-5. [DOI] [PubMed] [Google Scholar]
  52. Trelstad R. L. Mesenchymal cell polarity and morphogenesis of chick cartilage. Dev Biol. 1977 Sep;59(2):153–163. doi: 10.1016/0012-1606(77)90250-0. [DOI] [PubMed] [Google Scholar]
  53. Vega-Salas D. E., Salas P. J., Gundersen D., Rodriguez-Boulan E. Formation of the apical pole of epithelial (Madin-Darby canine kidney) cells: polarity of an apical protein is independent of tight junctions while segregation of a basolateral marker requires cell-cell interactions. J Cell Biol. 1987 Apr;104(4):905–916. doi: 10.1083/jcb.104.4.905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Virtanen I., Lehto V. P., Lehtonen E., Vartio T., Stenman S., Kurki P., Wager O., Small J. V., Dahl D., Badley R. A. Expression of intermediate filaments in cultured cells. J Cell Sci. 1981 Aug;50:45–63. doi: 10.1242/jcs.50.1.45. [DOI] [PubMed] [Google Scholar]
  55. Yang J., Richards J., Bowman P., Guzman R., Enami J., McCormick K., Hamamoto S., Pitelka D., Nandi S. Sustained growth and three-dimensional organization of primary mammary tumor epithelial cells embedded in collagen gels. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3401–3405. doi: 10.1073/pnas.76.7.3401. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES