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Analysis of desmosomal intramembrane particle populations and cytoskeletal elements: Detergent extraction and freeze-fracture

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Summary

We have examined sections and freeze-fracture replicas of Triton X-100 detergent-extracted desmosomes from murine palatal epithelium. After extraction of lipids as well as soluble proteins, a cytoskeletal framework remained which consisted of intermediate filaments, microfilaments, and intact desmosomal skeletons. Traversing filaments, which link the intermediate filaments to large intramembrane particles of the P-face, appeared undisturbed within the desmosomal skeletons. Compared to unextracted controls, extracted specimens displayed P- and E-face desmosomal intramembrane particles which were more fully exposed. A broad range of sizes and shapes was apparent for the P-face associated particles. E-face particles, some of which were exposed for the first time, were more homogeneous and generally smaller. Statistical data gathered from a large sample of P- and E-face particle diameters disclosed significant differences among the populations of the two faces. Both fracture faces of extracted desmosomal domains displayed a residual surface upon which the exposed particles seemed to remain lodged. The newly revealed structural features are presented in an hypothetical molecular model which provides for both vertical and horizontal stabilization of desmosomal subcomponents. The model may ultimately be relatable to emerging biochemical characterization.

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References

  • Arnn J, Staehelin LA (1981) The structure and function of spot desmosomes. Int J Dermatol 20:330–339

    Google Scholar 

  • Bennett V (1982) The molecular basis for membrane-cytoskeletal association in human erythrocytes. J Cell Biochem 18:49–65

    Google Scholar 

  • Borysenko JZ, Revel JP (1973) Experimental manipulation of desmosome structure. Am J Anat 137:403–422

    Google Scholar 

  • Branton D, Cohen CM, Tyler J (1981) Interaction of cytoskeletal proteins on the human erythrocyte membrane. Cell 24:24–32

    Google Scholar 

  • Breathnach AS, Stolinski C, Gross M (1972) Ultrastructure of foetal and postnatal human skin as revealed by the freeze-fracture replication technique. Micron 3:287–304

    Google Scholar 

  • Breathnach AS, Goodman T, Stolinski C, Gross M (1973) Freeze-fracture replication of cells of stratum corneum of human epidermis. J Anat 114:65–81

    Google Scholar 

  • Breathnach AS, Gross M, Martin B, Stolinski C (1976) A comparison of membrane fracture faces of fixed and unfixed glycerinated tissue. J Cell Sci 21:437–448

    Google Scholar 

  • Bretscher MS (1971) A major protein which spans the human erythrocyte membrane. J Mol Biol 59:351–357

    Google Scholar 

  • Bretscher MS (1973) Membrane structure: some general principles. Science 181:622–629

    Google Scholar 

  • Burridge K, Connell L (1983) Talin: a cytoskeletal component concentrated in adhesion plaques and other sites of actin-membrane interaction. Cell Motil 3:405–417

    Google Scholar 

  • Burridge K, Kelly T, Mangeat P (1982) Nonerythrocyte spectrin: actin-membrane attachment proteins occuring in many cell types. J Cell Biol 95:478–486

    Google Scholar 

  • Capaldi RA, Green DE (1972) Membrane proteins and membrane structure. FEBS Lett 25:205–209

    Google Scholar 

  • Caputo R, Pelluchetti D (1977) The junctions of normal human epidermis. J Ultrastruct Res 61:44–61

    Google Scholar 

  • Cohen SM, Gorbsky G, Steinberg MS (1983) Immunochemical characterization of related families of glycoproteins in desmosomes. J Biol Chem 258:2621–2627

    Google Scholar 

  • Damsky CH, Knudsen KA, Buck CA (1982) Integral membrane glycoproteins related to cell-substratum adhesion in mammalian cells. J Cell Biochem 18:1–13

    Google Scholar 

  • Drochmans P, Freudenstein C, Wanson J-C, Laurent L, Keenan TW, Stadler J, Leloup R, Franke WW (1978) Structure and biochemical composition of desmosomes and tonofilaments isolated from calf muzzle epidermis. J Cell Biol 79:427–443

    Google Scholar 

  • Farquhar MG, Palade GE (1963) Junctional complexes in various epithelia. J Cell Biol 17:375–412

    Google Scholar 

  • Fawcett DW (1961) Intercellular bridges. Exp Cell Res 8:174–187

    Google Scholar 

  • Fey EG, Wan KM, Penman S (1984) Epithelial cytoskeletal frame work and nuclear matrix-intermediate filament scaffold: threedimensional organization and protein composition. J Cell Biol 98:1973–1984

    Google Scholar 

  • Franke WW, Kapprell H-P, Mueller H (1983) Isolation and symmetrical splitting of desmosomal structures in 9 M urea. Eur J Cell Biol 32:117–130

    Google Scholar 

  • Geiger B, Schmid E, Franke WW (1983) Spatial distribution of proteins specific for desmosomes and adherens junctions in epithelial cells demonstrated by double immunofluorescence microscopy. Differentiation 23:189–205

    Google Scholar 

  • Glenny JR, Glenny P (1983) Spectrin, Fodrin and TRW 260/240: a family of related proteins lining the plasma membrane. Motil 3:671–682

    Google Scholar 

  • Glenny JR Jr, Glenny P, Weber K (1982) F-actin binding and cross-linking properties of porcine brain fodrin, a spectrin related molecule. J Biol Chem 257:9781–9789

    Google Scholar 

  • Goodman SR Jr, Zagon IS, Kulikowski RR (1981) Identification of a spectrin-like protein in nonerythroid cells. Proc Natl Acad Sci USA 78:7570–7574

    Google Scholar 

  • Gorbsky G, Steinberg M (1981) Isolation of the intercellular glycoproteins of desmosomes. J Cell Biol 90:243–248

    Google Scholar 

  • Helenius A, Simons K (1975) Solubilization of membranes by detergents. Biochim Biophys Acta 415:29–79

    Google Scholar 

  • Jones BM, Heppinstall K (1984) Trifluoroperazine inhibits formation of adhesion plaque complexes and cell sorting in aggregates of fibroblasts. Cell Biol Int Rep 8:65–77

    Google Scholar 

  • Kelly DE (1966) Fine structure of desmosomes and hemidesmosomes and an adepidermal globular layer in developing newt epidermis. J Cell Biol 28:51–72

    Google Scholar 

  • Kelly DE, Kuda AM (1981) Traversing filaments in desmosomal and hemidesmosomal attachments: freeze-fracture approaches toward their characterization. Anat Rec 199:1–14

    Google Scholar 

  • Kelly DE, Shienvold FL (1976) The desmosome: fine structural studies with freeze-fracture replication and tannic acid staining of sectioned epidermis. Cell Tissue Res 172:309–323

    Google Scholar 

  • Koch GLE (1981) The anchorage of cell surface receptors to the cytoskeleton. In: Schweiger HG (ed) International cell biology. Springer, Berlin New York, pp 321–330

    Google Scholar 

  • Lazarides E (1980) Intermediate filaments as mechanical integrators of cellular space. Nature 283:249–256

    Google Scholar 

  • LeLoup R, Lauret L, Ronveaux MF, Drochmans P, Wanson JC (1979) Desmosomes and desmogenesis in the epidermis of calf muzzle. Biol Cell 34:137–152

    Google Scholar 

  • Lux SE (1979) Spectrin-actin membrane skeleton of normal and abnormal blood cells. Semin Hematol 16:21–51

    Google Scholar 

  • Matoltsy AG (1975) Desmosomes, filaments and keratohyalin granules: their role in the stabilization and keratinization of the epidermis. J Invest Derm 65:127–142

    Google Scholar 

  • McNutt NS (1977) Freeze-fracture techniques and applications to the structural analysis of the mammalian plasma membrane. Cell Surface Rev 3:75–126

    Google Scholar 

  • McNutt NS, Weinstein RS (1973) Membrane ultrastructure at mammalian intercellular junctions. Prog Biophys Mol Bio 26:45–101

    Google Scholar 

  • McNutt NS, Hershberg RA, Weinstein RS (1971) Further observations on the occurrence of nexuses in benign and malignant human cervical epithelium. J Cell Biol 51:805–825

    Google Scholar 

  • Mueller H, Franke WW (1983) Biochemical and immunological characterization of desmoplakins I and II, the major polypeptides of the desmosomal plaque. J Mol Bio 163:647–671

    Google Scholar 

  • Nicolson GL (1976) Transmembrane control of the receptors on normal and tumor cells. I. Cytoplasmic influence over cell surface components. Biochim Biophys Acta 457:57–108

    Google Scholar 

  • Nicolson GL, Poste G (1976) The cancer cell: dynamic aspects and modifications in cell-surface organization (part I). New Eng J Med 295:197–203

    Google Scholar 

  • Nicolson GL, Poste G (1976) The cancer cell: dynamic aspects and modifications in cell-surface organization (part II). New Eng J Med 295:253–258

    Google Scholar 

  • Overton J (1982) Inhibition of desmosome formation with tunicamycin and with lectin in corneal cell aggregates. Dev Biol 92:66–72

    Google Scholar 

  • Repasky EA, Granger BL, Lazarides E (1982) Widespread occurrence of avian spectrin in nonerythroid cells. Cell 29:821–833

    Google Scholar 

  • Segrest JP, Kahane I, Jackson RL, Marchesi VT (1973) Major glycoprotein of the human erythrocyte membrane: evidence for an amphipathic molecular structure. Arch Biochem Biophys 155:167–183

    Google Scholar 

  • Shimono M, Clementi F (1977) Intercellular junctions of oral epithelium. II. Ultrastructural changes in rat buccal epithelium induced by trypsin digestion. J Ultrastruct Res 59:101–112

    Google Scholar 

  • Singer SJ, Nicolson GL (1972) The fluid mosaic model of the structure of cell membrane. Science 175:720–731

    Google Scholar 

  • Skerrow CJ (1978) Intercellular adhesion and its role in epidermal differentiation. Invest Cell Pathol 1:23–37

    Google Scholar 

  • Skerrow CJ (1979) Selective extraction of desmosomal proteins by low ionic strength media. Biochem Biophys Acta 579:241–245

    Google Scholar 

  • Skerrow CJ, Matoltsy AG (1974) Isolation of epidermal desmosomes. J Cell Biol 63:515–523

    Google Scholar 

  • Skerrow CJ, Matoltsy AG (1974) Chemical characterization of isolated epidermal desmosomes. J Cell Biol 63:524–530

    Google Scholar 

  • Sokal RR, Rohlf FJ (1981) Biometry: The principles and practice of statistics in biological research. WH Freeman and Company, San Francisco

    Google Scholar 

  • Staehelin LA (1974) Structure and function of intercellular junctions. Int Rev Cytol 39:191–283

    Google Scholar 

  • Staehelin LA, Hull BE (1978) Junctions between living cells. Sci Am 238:140–152

    Google Scholar 

  • Yu J, Fischman DA, Steck TL (1973) Selective solubilization of proteins and phospholipids from red blood cell membranes by nonionic detergents. J Supramol Struct 1:233–248

    Google Scholar 

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Authors and Affiliations

  1. Department of Anatomy and Cell Biology, University of Southern California, School of Medicine, Los Angeles, Calif., USA

    Mike Pirbazari &  Douglas E. Kelly

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  1. Mike Pirbazari
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  2. Douglas E. Kelly
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Pirbazari, M., Kelly, D.E. Analysis of desmosomal intramembrane particle populations and cytoskeletal elements: Detergent extraction and freeze-fracture. Cell Tissue Res. 241, 341–351 (1985). https://doi.org/10.1007/BF00217179

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  • DOI: https://doi.org/10.1007/BF00217179

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