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

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Affiliations

  • 1. Department of Anatomy and Cellular Biology, Harvard Medical School, Boston, Massachusetts 02115.
      Authors
    • Zuk A 1
    (1 author)

The Journal of Cell Biology, 01 Mar 1989, 108(3):903-919
https://doi.org/10.1083/jcb.108.3.903 PMID: 2537838 PMCID: PMC2115371

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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.

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J Cell Biol. 1989 Mar 1; 108(3): 903–919.
PMCID: PMC2115371
PMID: 2537838

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

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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.

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Selected References

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  • Abercrombie M, Heaysman JE, Pegrum SM. 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. [Abstract] [Google Scholar]
  • Bard JB, Hay ED. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Bergmann JE, Kupfer A, Singer SJ. Membrane insertion at the leading edge of motile fibroblasts. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1367–1371. [Europe PMC free article] [Abstract] [Google Scholar]
  • Caplan MJ, Stow JL, Newman AP, Madri J, Anderson HC, Farquhar MG, Palade GE, Jamieson JD. Dependence on pH of polarized sorting of secreted proteins. Nature. 1987 Oct 15;329(6140):632–635. [Abstract] [Google Scholar]
  • Cereijido M, Robbins ES, Dolan WJ, Rotunno CA, Sabatini DD. Polarized monolayers formed by epithelial cells on a permeable and translucent support. J Cell Biol. 1978 Jun;77(3):853–880. [Europe PMC free article] [Abstract] [Google Scholar]
  • Dodson JW, Hay ED. 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. [Abstract] [Google Scholar]
  • Elsdale T, Bard J. Collagen substrata for studies on cell behavior. J Cell Biol. 1972 Sep;54(3):626–637. [Europe PMC free article] [Abstract] [Google Scholar]
  • Fitchett JE, Hay ED. Medial edge epithelium transforms to mesenchyme after embryonic palatal shelves fuse. Dev Biol. 1989 Feb;131(2):455–474. [Abstract] [Google Scholar]
  • Franke WW, Schmid E, Breitkreutz D, Lüder M, Boukamp P, Fusenig NE, 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. [Abstract] [Google Scholar]
  • Franke WW, Grund C, Kuhn C, Jackson BW, 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. [Abstract] [Google Scholar]
  • Fuller S, von Bonsdorff CH, 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. [Abstract] [Google Scholar]
  • Garcia-Perez A, Smith WL. Use of monoclonal antibodies to isolate cortical collecting tubule cells: AVP induces PGE release. Am J Physiol. 1983 Mar;244(3):C211–C220. [Abstract] [Google Scholar]
  • Glenney JR, Jr, Glenney P. Comparison of spectrin isolated from erythroid and non-erythroid sources. Eur J Biochem. 1984 Nov 2;144(3):529–539. [Abstract] [Google Scholar]
  • Greenburg G, Hay ED. Epithelia suspended in collagen gels can lose polarity and express characteristics of migrating mesenchymal cells. J Cell Biol. 1982 Oct;95(1):333–339. [Europe PMC free article] [Abstract] [Google Scholar]
  • Greenburg G, Hay ED. 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. [Abstract] [Google Scholar]
  • Greenburg G, Hay ED. Cytoskeleton and thyroglobulin expression change during transformation of thyroid epithelium to mesenchyme-like cells. Development. 1988 Mar;102(3):605–622. [Abstract] [Google Scholar]
  • Haeuptle MT, Suard YL, Bogenmann E, Reggio H, Racine L, Kraehenbuhl JP. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Harris AS, Anderson JP, Yurchenco PD, Green LA, Ainger KJ, Morrow JS. Mechanisms of cytoskeletal regulation: functional and antigenic diversity in human erythrocyte and brain beta spectrin. J Cell Biochem. 1986;30(1):51–69. [Abstract] [Google Scholar]
  • Herzlinger DA, Easton TG, Ojakian GK. The MDCK epithelial cell line expresses a cell surface antigen of the kidney distal tubule. J Cell Biol. 1982 May;93(2):269–277. [Europe PMC free article] [Abstract] [Google Scholar]
  • Kaye GI, Wheeler HO, Whitlock RT, Lane N. Fluid transport in the rabbit gallbladder. A combined physiological and electron microscopic study. J Cell Biol. 1966 Aug;30(2):237–268. [Europe PMC free article] [Abstract] [Google Scholar]
  • Kupfer A, Kronebusch PJ, Rose JK, Singer SJ. A critical role for the polarization of membrane recycling in cell motility. Cell Motil Cytoskeleton. 1987;8(2):182–189. [Abstract] [Google Scholar]
  • Kupfer A, Louvard D, Singer SJ. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Lawrence JB, Singer RH. Intracellular localization of messenger RNAs for cytoskeletal proteins. Cell. 1986 May 9;45(3):407–415. [Abstract] [Google Scholar]
  • 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Matlin KS. 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. [Abstract] [Google Scholar]
  • Matlin KS, 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. [Abstract] [Google Scholar]
  • Matlin KS, 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • McAteer JA, Evan AP, Gardner KD. Morphogenetic clonal growth of kidney epithelial cell line MDCK. Anat Rec. 1987 Mar;217(3):229–239. [Abstract] [Google Scholar]
  • McDonald JA, Broekelmann TJ, Matheke ML, 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Misfeldt DS, Hamamoto ST, Pitelka DR. Transepithelial transport in cell culture. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1212–1216. [Europe PMC free article] [Abstract] [Google Scholar]
  • Mueller H, Franke WW. 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. [Abstract] [Google Scholar]
  • Nelson WJ, Veshnock PJ. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Nelson WJ, Veshnock PJ. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Osborn M, Weber K. Tumor diagnosis by intermediate filament typing: a novel tool for surgical pathology. Lab Invest. 1983 Apr;48(4):372–394. [Abstract] [Google Scholar]
  • Pasdar M, Nelson WJ. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Pasdar M, Nelson WJ. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Ramaekers FC, Osborn M, Schimid E, Weber K, Bloemendal H, Franke WW. Identification of the cytoskeletal proteins in lens-forming cells, a special epitheloid cell type. Exp Cell Res. 1980 Jun;127(2):309–327. [Abstract] [Google Scholar]
  • Richardson JC, Scalera V, Simmons NL. Identification of two strains of MDCK cells which resemble separate nephron tubule segments. Biochim Biophys Acta. 1981 Feb 18;673(1):26–36. [Abstract] [Google Scholar]
  • Rindler MJ, Chuman LM, Shaffer L, Saier MH., Jr Retention of differentiated properties in an established dog kidney epithelial cell line (MDCK). J Cell Biol. 1979 Jun;81(3):635–648. [Europe PMC free article] [Abstract] [Google Scholar]
  • Rindler MJ, Ivanov IE, Sabatini DD. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Rodriguez Boulan E, Sabatini DD. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Rodriguez-Boulan E, Paskiet KT, Sabatini DD. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Rogalski AA, Bergmann JE, Singer SJ. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Salas PJ, Misek DE, Vega-Salas DE, 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Schmid E, Tapscott S, Bennett GS, Croop J, Fellini SA, Holtzer H, Franke WW. Differential location of different types of intermediate-sized filaments in various tissues of the chicken embryo. Differentiation. 1979;15(1):27–40. [Abstract] [Google Scholar]
  • Simons K, Fuller SD. Cell surface polarity in epithelia. Annu Rev Cell Biol. 1985;1:243–288. [Abstract] [Google Scholar]
  • Smith ZD, Caplan MJ, Jamieson JD. Immunocytochemical localization of plasmalemmal proteins in semi-thin sections of epithelial monolayers. J Histochem Cytochem. 1988 Mar;36(3):311–316. [Abstract] [Google Scholar]
  • Stevenson BR, Siliciano JD, Mooseker MS, Goodenough DA. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Tomasek JJ, Hay ED. 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Tomasek JJ, Hay ED, 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. [Abstract] [Google Scholar]
  • Trelstad RL. Mesenchymal cell polarity and morphogenesis of chick cartilage. Dev Biol. 1977 Sep;59(2):153–163. [Abstract] [Google Scholar]
  • Vega-Salas DE, Salas PJ, 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Virtanen I, Lehto VP, Lehtonen E, Vartio T, Stenman S, Kurki P, Wager O, Small JV, Dahl D, Badley RA. Expression of intermediate filaments in cultured cells. J Cell Sci. 1981 Aug;50:45–63. [Abstract] [Google Scholar]
  • 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. [Europe PMC free article] [Abstract] [Google Scholar]
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