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Abstract 


Actin, keratin, vinculin and desmoplakin organization were studied in primary mouse keratinocytes before and during Ca2+-induced cell contact formation. Double-label fluorescence shows that in cells cultured in low Ca2+ medium, keratin-containing intermediate filament bundles (IFB) and desmoplakin-containing spots are both concentrated towards the cell center in a region bounded by a series of concentric microfilament bundles (MFB). Within 5-30 min after raising Ca2+ levels, a discontinuous actin/vinculin-rich, submembranous zone of fluorescence appears at cell-cell interfaces. This zone is usually associated with short, perpendicular MFB, which become wider and longer with time. Later, IFB and the desmoplakin spots are seen aligned along the perpendicular MFB as they become redistributed to cell-cell interfaces where desmosomes form. Ultrastructural analysis confirms that before the Ca2+ switch, IFB and desmosomal components are found predominantly within the perimeter defined by the outermost of the concentric MFB. Individual IF often splay out, becoming interwoven into these MFB in the region of cell-substrate contact. In the first 30 min after the Ca2+ switch, areas of submembranous dense material (identified as adherens junctions), which are associated with the perpendicular MFB, can be seen at newly formed cell-cell contact sites. By 1-2 h, IFB-desmosomal component complexes are aligned with the perpendicular MFB as the complexes become redistributed to cell-cell interfaces. Cytochalasin D treatment causes the redistribution of actin into numerous patches; keratin-containing IFB undergo a concomitant redistribution, forming foci that coincide with the actin-containing aggregates. These results are consistent with an IF-MF association before and during desmosome formation in the primary mouse epidermal keratinocyte culture system, and with the temporal and spatial coordination of desmosome and adherens junction formation.

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J Cell Biol. 1987 May 1; 104(5): 1389–1402.
PMCID: PMC2114486
PMID: 2437129

The relationship between intermediate filaments and microfilaments before and during the formation of desmosomes and adherens-type junctions in mouse epidermal keratinocytes

Abstract

Actin, keratin, vinculin and desmoplakin organization were studied in primary mouse keratinocytes before and during Ca2+-induced cell contact formation. Double-label fluorescence shows that in cells cultured in low Ca2+ medium, keratin-containing intermediate filament bundles (IFB) and desmoplakin-containing spots are both concentrated towards the cell center in a region bounded by a series of concentric microfilament bundles (MFB). Within 5-30 min after raising Ca2+ levels, a discontinuous actin/vinculin-rich, submembranous zone of fluorescence appears at cell-cell interfaces. This zone is usually associated with short, perpendicular MFB, which become wider and longer with time. Later, IFB and the desmoplakin spots are seen aligned along the perpendicular MFB as they become redistributed to cell-cell interfaces where desmosomes form. Ultrastructural analysis confirms that before the Ca2+ switch, IFB and desmosomal components are found predominantly within the perimeter defined by the outermost of the concentric MFB. Individual IF often splay out, becoming interwoven into these MFB in the region of cell-substrate contact. In the first 30 min after the Ca2+ switch, areas of submembranous dense material (identified as adherens junctions), which are associated with the perpendicular MFB, can be seen at newly formed cell-cell contact sites. By 1-2 h, IFB- desmosomal component complexes are aligned with the perpendicular MFB as the complexes become redistributed to cell-cell interfaces. Cytochalasin D treatment causes the redistribution of actin into numerous patches; keratin-containing IFB undergo a concomitant redistribution, forming foci that coincide with the actin-containing aggregates. These results are consistent with an IF-MF association before and during desmosome formation in the primary mouse epidermal keratinocyte culture system, and with the temporal and spatial coordination of desmosome and adherens junction formation.

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Selected References

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  • Bloom GS, Vallee RB. Association of microtubule-associated protein 2 (MAP 2) with microtubules and intermediate filaments in cultured brain cells. J Cell Biol. 1983 Jun;96(6):1523–1531. [Europe PMC free article] [Abstract] [Google Scholar]
  • Celis JE, Small JV, Larsen PM, Fey SJ, De Mey J, Celis A. Intermediate filaments in monkey kidney TC7 cells: focal centers and interrelationship with other cytoskeletal systems. Proc Natl Acad Sci U S A. 1984 Feb;81(4):1117–1121. [Europe PMC free article] [Abstract] [Google Scholar]
  • Cooke P. A filamentous cytoskeleton in vertebrate smooth muscle fibers. J Cell Biol. 1976 Mar;68(3):539–556. [Europe PMC free article] [Abstract] [Google Scholar]
  • Cowin P, Kapprell HP, Franke WW, Tamkun J, Hynes RO. Plakoglobin: a protein common to different kinds of intercellular adhering junctions. Cell. 1986 Sep 26;46(7):1063–1073. [Abstract] [Google Scholar]
  • FARQUHAR MG, PALADE GE. Junctional complexes in various epithelia. J Cell Biol. 1963 May;17:375–412. [Europe PMC free article] [Abstract] [Google Scholar]
  • Fawcett DW, McNutt NS. The ultrastructure of the cat myocardium. I. Ventricular papillary muscle. J Cell Biol. 1969 Jul;42(1):1–45. [Europe PMC free article] [Abstract] [Google Scholar]
  • Geiger B. A 130K protein from chicken gizzard: its localization at the termini of microfilament bundles in cultured chicken cells. Cell. 1979 Sep;18(1):193–205. [Abstract] [Google Scholar]
  • Geiger B. Involvement of vinculin in contact-induced cytoskeletal interactions. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 2):671–682. [Abstract] [Google Scholar]
  • Geiger B. Membrane-cytoskeleton interaction. Biochim Biophys Acta. 1983 Aug 11;737(3-4):305–341. [Abstract] [Google Scholar]
  • Geiger B, Dutton AH, Tokuyasu KT, Singer SJ. Immunoelectron microscope studies of membrane-microfilament interactions: distributions of alpha-actinin, tropomyosin, and vinculin in intestinal epithelial brush border and chicken gizzard smooth muscle cells. J Cell Biol. 1981 Dec;91(3 Pt 1):614–628. [Europe PMC free article] [Abstract] [Google Scholar]
  • Geiger B, Schmid E, Franke WW. Spatial distribution of proteins specific for desmosomes and adhaerens junctions in epithelial cells demonstrated by double immunofluorescence microscopy. Differentiation. 1983;23(3):189–205. [Abstract] [Google Scholar]
  • Geiger B, Singer SJ. Association of microtubules and intermediate filaments in chicken gizzard cells as detected by double immunofluorescence. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4769–4773. [Europe PMC free article] [Abstract] [Google Scholar]
  • Geiger B, Tokuyasu KT, Dutton AH, Singer SJ. Vinculin, an intracellular protein localized at specialized sites where microfilament bundles terminate at cell membranes. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4127–4131. [Europe PMC free article] [Abstract] [Google Scholar]
  • Geiger B, Volk T, Volberg T. Molecular heterogeneity of adherens junctions. J Cell Biol. 1985 Oct;101(4):1523–1531. [Europe PMC free article] [Abstract] [Google Scholar]
  • Green KJ, Talian JC, Goldman RD. Relationship between intermediate filaments and microfilaments in cultured fibroblasts: evidence for common foci during cell spreading. Cell Motil Cytoskeleton. 1986;6(4):406–418. [Abstract] [Google Scholar]
  • Griffith LM, Pollard TD. The interaction of actin filaments with microtubules and microtubule-associated proteins. J Biol Chem. 1982 Aug 10;257(15):9143–9151. [Abstract] [Google Scholar]
  • Hennings H, Holbrook KA. Calcium regulation of cell-cell contact and differentiation of epidermal cells in culture. An ultrastructural study. Exp Cell Res. 1983 Jan;143(1):127–142. [Abstract] [Google Scholar]
  • Hennings H, Michael D, Cheng C, Steinert P, Holbrook K, Yuspa SH. Calcium regulation of growth and differentiation of mouse epidermal cells in culture. Cell. 1980 Jan;19(1):245–254. [Abstract] [Google Scholar]
  • Hirokawa N, Cheney RE, Willard M. Location of a protein of the fodrin-spectrin-TW260/240 family in the mouse intestinal brush border. Cell. 1983 Mar;32(3):953–965. [Abstract] [Google Scholar]
  • Hubbard BD, Lazarides E. Copurification of actin and desmin from chicken smooth muscle and their copolymerization in vitro to intermediate filaments. J Cell Biol. 1979 Jan;80(1):166–182. [Europe PMC free article] [Abstract] [Google Scholar]
  • Hull BE, Staehelin LA. The terminal web. A reevaluation of its structure and function. J Cell Biol. 1979 Apr;81(1):67–82. [Europe PMC free article] [Abstract] [Google Scholar]
  • Jackson BW, Grund C, Schmid E, Bürki K, Franke WW, Illmensee K. Formation of cytoskeletal elements during mouse embryogenesis. Intermediate filaments of the cytokeratin type and desmosomes in preimplantation embryos. Differentiation. 1980;17(3):161–179. [Abstract] [Google Scholar]
  • Jockusch BM, Isenberg G. Interaction of alpha-actinin and vinculin with actin: opposite effects on filament network formation. Proc Natl Acad Sci U S A. 1981 May;78(5):3005–3009. [Europe PMC free article] [Abstract] [Google Scholar]
  • Jones JC, Goldman RD. Intermediate filaments and the initiation of desmosome assembly. J Cell Biol. 1985 Aug;101(2):506–517. [Europe PMC free article] [Abstract] [Google Scholar]
  • Jones JC, Goldman AE, Steinert PM, Yuspa S, Goldman RD. Dynamic aspects of the supramolecular organization of intermediate filament networks in cultured epidermal cells. Cell Motil. 1982;2(3):197–213. [Abstract] [Google Scholar]
  • Jones JC, Goldman AE, Yang HY, Goldman RD. The organizational fate of intermediate filament networks in two epithelial cell types during mitosis. J Cell Biol. 1985 Jan;100(1):93–102. [Europe PMC free article] [Abstract] [Google Scholar]
  • Kartenbeck J, Schmid E, Franke WW, Geiger B. Different modes of internalization of proteins associated with adhaerens junctions and desmosomes: experimental separation of lateral contacts induces endocytosis of desmosomal plaque material. EMBO J. 1982;1(6):725–732. [Europe PMC free article] [Abstract] [Google Scholar]
  • Knapp LW, O'Guin WM, Sawyer RH. Drug-induced alterations of cytokeratin organization in cultured epithelial cells. Science. 1983 Feb 4;219(4584):501–503. [Abstract] [Google Scholar]
  • Knapp LW, O'Guin WM, Sawyer RH. Rearrangement of the keratin cytoskeleton after combined treatment with microtubule and microfilament inhibitors. J Cell Biol. 1983 Dec;97(6):1788–1794. [Europe PMC free article] [Abstract] [Google Scholar]
  • Leterrier JF, Liem RK, Shelanski ML. Interactions between neurofilaments and microtubule-associated proteins: a possible mechanism for intraorganellar bridging. J Cell Biol. 1982 Dec;95(3):982–986. [Europe PMC free article] [Abstract] [Google Scholar]
  • Mattey DL, Garrod DR. Calcium-induced desmosome formation in cultured kidney epithelial cells. J Cell Sci. 1986 Sep;85:95–111. [Abstract] [Google Scholar]
  • OVERTON J. Desmosome development in normal and reassociating cells in the early chick blastoderm. Dev Biol. 1962 Jun;4:532–548. [Abstract] [Google Scholar]
  • Overton J. Formation of junctions and cell sorting in aggregates of chick and mouse cells. Dev Biol. 1977 Jan;55(1):103–116. [Abstract] [Google Scholar]
  • Pankov RG, Uschewa AA, Tasheva BT, Petrov PT, Markov GG. Actin participates in the structure of liver intermediate filaments. Cell Biol Int Rep. 1985 Nov;9(11):1003–1011. [Abstract] [Google Scholar]
  • Runge MS, Laue TM, Yphantis DA, Lifsics MR, Saito A, Altin M, Reinke K, Williams RC., Jr ATP-induced formation of an associated complex between microtubules and neurofilaments. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1431–1435. [Europe PMC free article] [Abstract] [Google Scholar]
  • Sattilaro RF, Dentler WL, LeCluyse EL. Microtubule-associated proteins (MAPs) and the organization of actin filaments in vitro. J Cell Biol. 1981 Aug;90(2):467–473. [Europe PMC free article] [Abstract] [Google Scholar]
  • Schliwa M. Action of cytochalasin D on cytoskeletal networks. J Cell Biol. 1982 Jan;92(1):79–91. [Europe PMC free article] [Abstract] [Google Scholar]
  • Schloss JA, Milsted A, Goldman RD. Myosin subfragment binding for the localization of actin-like microfilaments in cultured cells. A light and electron microscope study. J Cell Biol. 1977 Sep;74(3):794–815. [Europe PMC free article] [Abstract] [Google Scholar]
  • Starger JM, Brown WE, Goldman AE, Goldman RD. Biochemical and immunological analysis of rapidly purified 10-nm filaments from baby hamster kidney (BHK-21) cells. J Cell Biol. 1978 Jul;78(1):93–109. [Europe PMC free article] [Abstract] [Google Scholar]
  • Trelstad RL, Hay ED, Revel JD. Cell contact during early morphogenesis in the chick embryo. Dev Biol. 1967 Jul;16(1):78–106. [Abstract] [Google Scholar]
  • Volk T, Geiger B. A 135-kd membrane protein of intercellular adherens junctions. EMBO J. 1984 Oct;3(10):2249–2260. [Europe PMC free article] [Abstract] [Google Scholar]
  • Volk T, Geiger B. A-CAM: a 135-kD receptor of intercellular adherens junctions. I. Immunoelectron microscopic localization and biochemical studies. J Cell Biol. 1986 Oct;103(4):1441–1450. [Europe PMC free article] [Abstract] [Google Scholar]
  • Volk T, Geiger B. A-CAM: a 135-kD receptor of intercellular adherens junctions. II. Antibody-mediated modulation of junction formation. J Cell Biol. 1986 Oct;103(4):1451–1464. [Europe PMC free article] [Abstract] [Google Scholar]
  • Volberg T, Geiger B, Kartenbeck J, Franke WW. Changes in membrane-microfilament interaction in intercellular adherens junctions upon removal of extracellular Ca2+ ions. J Cell Biol. 1986 May;102(5):1832–1842. [Europe PMC free article] [Abstract] [Google Scholar]
  • Watt FM, Mattey DL, Garrod DR. Calcium-induced reorganization of desmosomal components in cultured human keratinocytes. J Cell Biol. 1984 Dec;99(6):2211–2215. [Europe PMC free article] [Abstract] [Google Scholar]
  • Yuspa SH, Harris CC. Altered differentiation of mouse epidermal cells treated with retinyl acetate in vitro. Exp Cell Res. 1974 May;86(1):95–105. [Abstract] [Google Scholar]

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