Electron microscopy of proteoglycans

  • C. Johan
  • O. Thyberg
Part of the Electron Microscopy in Biology and Medicine book series (EMBM, volume 3)


In addition to the fibrillar proteins collagen and elastin, proteoglycans constitute a major macro-molecular component in the extracellular matrix of connective tissues. They have also been identified as components of basement membranes and cell surface coats. Chemically, the proteoglycans consist of a protein core to which glycosaminoglycan chains are covalently attached. The latter are composed of alternating uronic acid and hexosamine residues and are all polyanions with acidic sulfate and/or carboxyl groups (1). A particularly high content of proteoglycans is found in cartilage and much of the information that is available today derives from studies on this tissue. An average cartilage proteoglycan has a molecular weight of about 2.5 af09106 and is composed of a protein core to which about 100 chon-droitin sulfate and 50–60 keratan sulfate chains are bound. Within the cartilaginous matrix, most of these monomers occur in large aggregates, formed by noncovalent interaction with hyaluronic acid and link proteins (2–4). The structure of proteoglycans in other connective tissues, in basement membranes, and in cell surface coats is less well known. The type, number, and size of glycosaminoglycan chains per molecule have been found to vary considerably, but the supramolecular organization (e.g. aggregate formation) is still poorly defined.


Hyaluronic Acid Collagen Fibril Alcian Blue Hyaline Cartilage Cartilage Proteoglycan 
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  1. 1.
    Lindahl U, Höök M: Glycosaminoglycans and their binding to biological macromolecules. Ann Rev Biochem 47: 385–417, 1978.PubMedCrossRefGoogle Scholar
  2. 2.
    Hascall VC, Heinegård DK: Structure of cartilage proteoglycans. In: Glycoconjugate Research, vol 1. Gregory JD, Jeanloz RW (eds), New York, Academic Press, 1979, pp 341–374.Google Scholar
  3. 3.
    Hascall VC, Hascall GK: Proteoglycans. In: Cell Biology of Extracellular Matrix. Hay ED (ed), New York, Plenum, 1981, pp 39–63.CrossRefGoogle Scholar
  4. 4.
    Hascall VC, Kimura JH: Proteoglycans: isolation and characterization. Methods Enzymol 82: 769–800, 1982.PubMedCrossRefGoogle Scholar
  5. 5.
    Slavkin HC, Greulich RC (eds): Extracellular Matrix Influences on Gene Expression. New York, Academic Press, 1975.Google Scholar
  6. 6.
    Lash JW, Burger MM (eds): Cell and Tissue Interactions. Society of general physiologists series 32, New York, Raven Press, 1977.Google Scholar
  7. 7.
    Lennarz WJ (ed): The Biochemistry of Glycoproteins and Proteoglycans. New York, Plenum Press, 1980.Google Scholar
  8. 8.
    Godman GC, Lane N: On the site of sulfation in the chondrocyte. J Cell Biol 21: 353–366, 1964.PubMedCrossRefGoogle Scholar
  9. 9.
    Fewer D, Threadgold J, Sheldon H: Studies on cartilage. V. Electron microscopic observations on the autoradiographic localization of S35 in cells and matrix. J Ultrastruct Res 11: 166–172, 1964.PubMedCrossRefGoogle Scholar
  10. 10.
    Curran RC, Clark AE, Lovell D: Acid mucopolysaccharides in electron microscopy. The use of the colloidal iron method. J Anat 99: 427–434, 1965.PubMedGoogle Scholar
  11. 11.
    Revel J-P: A stain for the ultrastructural localization of acid mucopolysaccharides. J Microsc 3: 535–544, 1964.Google Scholar
  12. 12.
    Luft JH: Ruthenium red and violet. I. Chemistry, purification, methods of use for electron microscopy and mechanism of action. Anat Rec 171: 347–368, 1971.PubMedCrossRefGoogle Scholar
  13. 13.
    Luft JH: Ruthenium red and violet. II. Fine structural localization in animal tissues. Anat Rec 171: 369–416, 1971.PubMedCrossRefGoogle Scholar
  14. 14.
    Behnke O, Zelander T: Preservation of intercellular substances by the cationic dye alcian blue in preparative procedures for electron microscopy. J Ultrastruct Res 31: 424–438, 1970.PubMedCrossRefGoogle Scholar
  15. 15.
    Khan TA, Overton J: Lanthanum staining of developing chick cartilage and reaggregating cartilage cells. J Cell Biol 44: 433–438, 1970.PubMedCrossRefGoogle Scholar
  16. 16.
    Eisenstein R, Arsenis C, Kuettner KE: Electron microscopic studies of cartilage matrix using lysozyme as a vital stain. J Cell Biol 46: 626–631, 1970.PubMedCrossRefGoogle Scholar
  17. 17.
    Shepard N, Mitchell N: The localization of proteoglycan by light and electron microscopy using safranin O. A study of epiphyseal cartilage. J Ultrastruct Res 54: 451–460, 1976.PubMedCrossRefGoogle Scholar
  18. 18.
    Shepard N, Mitchell N: Simultaneous localization of proteoglycan by light and electron microscopy using toluidine blue O. A study of epiphyseal cartilage. J Histochem Cytochem 24: 621–629, 1976.PubMedCrossRefGoogle Scholar
  19. 19.
    Scott JE, Orford CR, Hughes EW: Proteoglycan–collagen arrangements in developing rat tail tendon. An electron-microscopical and biochemical investigation. Biochem J 195: 573–581, 1981.PubMedGoogle Scholar
  20. 20.
    Takagi M, Parmley RT, Denys FR: Ultrastructural cytochemistry and radioautography of complex carbohydrates in secretory granules of epiphyseal chondrocytes. Lab Invest 44: 116–126, 1981.PubMedGoogle Scholar
  21. 21.
    Scott JE: Histochemistry of alcian blue. II. The structure of alcian blue 8GX. Histochemie 30: 215–234, 1972.PubMedGoogle Scholar
  22. 22.
    Scott JE: Histochemistry of alcian blue. III. The molecular basis of staining by alcian blue 8GX and analogous phthalocyanins. Histochemie 32: 191–212, 1972.PubMedCrossRefGoogle Scholar
  23. 23.
    Linker A, Hovingh P: The uses of degradative enzymes as tools for identification and structural analysis of glycosaminoglycans. Fed Proc 36: 43–46, 1977.PubMedGoogle Scholar
  24. 24.
    Schofield BH, Williams BR, Doty SB: Alcian blue staining of cartilage for electron microscopy. Application of the critical electrolyte concentration principle. Histochem J 7: 139–149, 1975.PubMedCrossRefGoogle Scholar
  25. 25.
    Hoshino M, Yamada K: Effects of digestion with chondroitinases upon mucosaccharide stainings of rabbit cartilage as revealed by electron microscopy. Histochemie 32: 221–229, 1972.PubMedCrossRefGoogle Scholar
  26. 26.
    Dorfman A, Vertel BM, Schwartz NB: Immunological methods in the study of chondroitin sulfate proteoglycans. Curr Top Dev Biol 14: 169–198, 1980.PubMedCrossRefGoogle Scholar
  27. 27.
    Poole AR, Pidoux I, Reiner A, Tang L–H, Choi H, Rosenberg L: Localization of proteoglycan monomer and link protein in the matrix of bovine articular cartilage. J Histochem Cytochem 28: 621–635, 1980.PubMedCrossRefGoogle Scholar
  28. 28.
    Mangkornkanok–Mark M, Eisenstein R, Bahu RM: Immunologic studies of bovine aortic and cartilage proteoglycans. J Histochem Cytochem 29: 547–552, 1981.PubMedCrossRefGoogle Scholar
  29. 29.
    Poole AR, Pidoux I, Reiner A, Rosenberg L: An immunoelectron microscope study of the organization of proteoglycan monomer, link protein, and collagen in the matrix of articular cartilage. J Cell Biol 93: 921–937, 1982.PubMedCrossRefGoogle Scholar
  30. 30.
    Dessau W, Vertel BM, von der Mark H, von der Mark K: Extracellular matrix formation by chondrocytes in monolayer culture. J Cell Biol 90: 78–83, 1981.PubMedCrossRefGoogle Scholar
  31. 31.
    Gonatas NK, Avrameas S: Detection of carbohydrates with lectin–peroxidase conjugates. In: Methods in Cell Biology vol XV. Prescott DM (ed), New York, Academic Press, 1977, pp 387–406.CrossRefGoogle Scholar
  32. 32.
    Godman GC, Porter KR: Chondrogenesis studied with the electron microscope. J Biophys Biochem Cytol 8: 719–760, 1960.PubMedCrossRefGoogle Scholar
  33. 33.
    Matukas VJ, Panner BJ, Orbison JL: Studies on ultrastructural identification and distribution of protein-polysaccharide in cartilage matrix. J Cell Biol 32: 365–377, 1967.PubMedCrossRefGoogle Scholar
  34. 34.
    Smith JW: The disposition of proteinpolysaccharide in the epiphyseal plate cartilage of the young rabbit. J Cell Sci 6: 843–864, 1970.PubMedGoogle Scholar
  35. 35.
    Anderson HC, Sajdera SW: The fine structure of bovine nasal cartilage. Extraction as a technique to study proteoglycans and collagen in cartilage matrix. J Cell Biol 49: 650–663, 1971.PubMedCrossRefGoogle Scholar
  36. 36.
    Thyberg J, Lohmander S, Friberg U: Electron microscopic demonstration of proteoglycans in guinea pig epiphyseal cartilage. J Ultrastruct Res 45: 407–427, 1973.PubMedCrossRefGoogle Scholar
  37. 37.
    Thyberg J, Nilsson S, Friberg U: Electron microscopic studies on guinea pig rib cartilage. Structural heterogeneity and effects of extraction with quanidine–HCI. Z Zellforsch Mikrosk Anat 146: 83–102, 1973.PubMedCrossRefGoogle Scholar
  38. 38.
    Ruggeri A, Dell’Orbo C, Quacci D: Electron microscopic visualization of proteoglycans with ruthenium red. Histochem J 9: 249–252, 1977.PubMedCrossRefGoogle Scholar
  39. 39.
    Hascall GK: Cartilage proteoglycans: comparison of sectioned and spread whole molecules. J Ultrastruct Res 70: 369–375, 1980.PubMedCrossRefGoogle Scholar
  40. 40.
    Myers DB, Highton TC, Rayns DG: Acid mucopolysaccharides closely associated with collagen fibrils in normal human synovium. J Ultrastruct Res 28: 203–213, 1969.PubMedCrossRefGoogle Scholar
  41. 41.
    Takusagawa K, Ariji F, Shida K, Sato T, Asoo N, Konno K: Electron microscopic observations on pulmonary connective tissue stained by ruthenium red. Histochem J 14: 257–271, 1982.PubMedCrossRefGoogle Scholar
  42. 42.
    Smith JW, Frame J: Observations on the collagen and proteinpolysaccharide complex of rabbit corneal stroma. J Cell Sci 4: 421–436, 1969.PubMedGoogle Scholar
  43. 43.
    Trelstad RL, Hayashi K, Toole BP: Epithelial collagens and glycosaminoglycans in the embryonic cornea. Macromolecular order and morphogenesis in the basement membrane. J Cell Biol 62: 815–830, 1974.PubMedCrossRefGoogle Scholar
  44. 44.
    Wight TN, Ross R: Proteoglycans in primate arteries. I. Ultrastructural localization and distribution in the intima. J Cell Biol 67: 660–674, 1975.PubMedCrossRefGoogle Scholar
  45. 45.
    Eisenstein R, Kuettner K: The ground substance of the arterial wall. Part 2. Electron-microscopic studies. Atherosclerosis 27: 37–46, 1976.CrossRefGoogle Scholar
  46. 46.
    Riva R, Marchini M, Strocchi R: Proteoglycans and their relationship with the other components of the rabbit aorta wall observed in two different experimental conditions. Acta Histochem 65: 233–242, 1979.PubMedGoogle Scholar
  47. 47.
    Oegema TR, Hascall VC, Eisenstein R: Characterization of bovine aorta proteoglycan extracted with guanidine hydrochloride in the presence of protease inhibitors. J Biol Chem 254: 1312–1318, 1979.PubMedGoogle Scholar
  48. 48.
    Salisbury BGJ, Wagner WD: Isolation and preliminary characterization of proteoglycans dissociatively extracted from human aorta. J Biol Chem 256: 8050–8057, 1981.PubMedGoogle Scholar
  49. 49.
    Schmidt A, Prager M, Selmke P, Buddecke E: Isolation and properties of proteoglycans from bovine aorta. Eur J Biochem 125: 95–101, 1982.PubMedCrossRefGoogle Scholar
  50. 50.
    Nygren H, Hansson H-A, Linde A: Ultrastructural localisation of proteoglycans in the odontoblast-predentin region of rat incisor. Cell Tissue Res 168: 277–287, 1976.PubMedCrossRefGoogle Scholar
  51. 51.
    Takagi M, Parmley RT, Denys FR: Ultrastructural localization of complex carbohydrates in odontoblasts, predentin, and dentin. J Histochem Cytochem 29: 747–758, 1981.PubMedCrossRefGoogle Scholar
  52. 52.
    Hay ED, Meier S: Glycosaminoglycan synthesis by embryonic inductors: neural tube, notochord, and lens. J Cell Biol 62: 889–898, 1974.PubMedCrossRefGoogle Scholar
  53. 53.
    Cohn RH, Banerjee SD, Bernfield MR: Basal lamina of embryonic salivary epithelia. Nature of glycosaminoglycan and organization of extracellular materials. J Cell Biol 73: 464–478, 1977.PubMedCrossRefGoogle Scholar
  54. 54.
    Kanwar YS, Fraquhar MG: Anionic sites in the glomerular basement membrane. In vivo and in vitro localization to the laminae rarae by cationic probes. J Cell Biol 81: 137–153, 1979.Google Scholar
  55. 55.
    Kanwar YS, Farquhar MG: Presence of heparan sulfate in the glomerular basement membrane. Proc Natl Acad Sci USA 76: 1303–1307, 1979.PubMedCrossRefGoogle Scholar
  56. 56.
    Kanwar YS, Hascall VC, Farquhar MG, Partial characterization of newly synthesized proteoglycans isolated from the glomerular basement membrane. J Cell Biol 90: 527–532, 1981.PubMedCrossRefGoogle Scholar
  57. 57.
    Kanwar YS, Linker A, Farquhar MG: Increased permeability of the glomerular basement membrane to ferritin after removal of glycosaminoglycans (heparan sulfate) by enzyme digestion. J Cell Biol 86: 688–693, 1980.PubMedCrossRefGoogle Scholar
  58. 58.
    Del Rosso M, Cappelletti R, Vannucchi S, Romagnani S, Chiarugi VP: Selective exposure of mucopolysaccharides is involved in macrophage physiology. Biochim Biophys Acta 586: 512–517, 1979.PubMedCrossRefGoogle Scholar
  59. 59.
    Vogel KG, Kendall VF: Cell-surface glycosaminoglycans: turnover in cultured human embryo fibroblasts (IMR-90). J Cell Physiol 103: 475–487, 1980.PubMedCrossRefGoogle Scholar
  60. 60.
    Kjellén L, Pettersson I, Höök M: Cell-surface heparan sulfate: An intercalated membrane proteoglycan. Proc Natl Acad Sci USA 78: 5371–5375, 1981.PubMedCrossRefGoogle Scholar
  61. 61.
    Simionescu N, Simionescu M: Galloylglucoses of low molecular weight as mordant in electron microscopy. I. Procedure, and evidence for mordanting effect. J Cell Biol 70: 608–621, 1976.PubMedCrossRefGoogle Scholar
  62. 62.
    Luft JH: The structure and properties of the cell surface coat. Int Rev Cytol 45: 291–382, 1976.PubMedCrossRefGoogle Scholar
  63. 63.
    Spicer SS, Baron DA, Sato A, Schulte BA: Variability of cell surface glycoconjugates — relation to differences in cell function. J Histochem Cytochem 29: 994–1002, 1981.PubMedCrossRefGoogle Scholar
  64. 64.
    Danon DL, Goldstein L, Marikovsky Y, Skutelsky E: Use of ca-tionized ferritin as a label of negative charges on cell surfaces. J Ultrastruct Res 38: 500–510, 1972.PubMedCrossRefGoogle Scholar
  65. 65.
    Simionescu M, Simionescu N, Silbert JE, Palade GE: Differentiated microdomains on the luminal surface of the capillary endothelium. II. Partial characterization of their anionic sites. J Cell Biol 90: 614–621, 1981.PubMedCrossRefGoogle Scholar
  66. 66.
    Kleinschmidt AK, Zahn RK: Uber Desoxyribonukleinsäure-Mole-kylen in Protein Mischfilmen. Z Naturforsch (B) 146: 770–779, 1959.Google Scholar
  67. 67.
    Rosenberg L, Hellmann W, Kleinschmidt AK: Macromolecular models of proteinpolysaccharides from bovine nasal cartilage based on electron microscopic studies. J Biol Chem 245: 4123–4130, 1970.PubMedGoogle Scholar
  68. 68.
    Lang D, Mitani M: Simplified quantitative electron microscopy of biopolymers. Biopolymers 9: 373–379, 1970.PubMedCrossRefGoogle Scholar
  69. 69.
    Thyberg J, Lohmander S, Heinegård D: Proteoglycans of hyaline cartilage. Electron-microscopic studies on isolated molecules. Bio-chem J 151: 157–166, 1975.Google Scholar
  70. 70.
    Kleinschmidt AK: Electron microscopic studies of macromolecules without appositional contrast. Philos Trans R Soc Lond [Biol] 261: 143–149, 1971.CrossRefGoogle Scholar
  71. 71.
    Panessa BJ, McCorkle RA, Hoffman P, Warren JB, Coleman G: Ultrastructure of hydrated proteoglycans using a pulsed plasma source. Ultramicroscopy 6: 139–148, 1981.PubMedGoogle Scholar
  72. 72.
    Fessier JH, Fessier LI: Electron microscopic visualization of the poly-saccharide hyaluronic acid. Proc Natl Acad Sci USA 56: 141–147, 1966.CrossRefGoogle Scholar
  73. 73.
    Wellauer P, Wyler T, Buddecke E: Electron microscopic and physico-chemical studies on bovine nasal cartilage proteoglycan. Hoppe Seylers Z Physiol Chem 353: 1043–1052, 1972.PubMedCrossRefGoogle Scholar
  74. 74.
    Yoneda M: Electron microscopic studies of proteoglycans from epiphyseal cartilage of suckling rats. Connect Tissue 9 (2): 1–12, 1977.Google Scholar
  75. 75.
    Buckwalter JA, Rosenberg LR: Electron microscopic studies of cartilage proteoglycans. Direct evidence for the variable length of the chondroitin sulfate-rich region of proteoglycan subunit core protein. J Biol Chem 257: 9830–9839, 1982.PubMedGoogle Scholar
  76. 76.
    Rosenberg L, Hellmann W, Kleinschmidt AK: Electron microscopic studies of proteoglycan aggregates from bovine articular cartilage. J Biol Chem 250: 1877–1883, 1975.PubMedGoogle Scholar
  77. 77.
    Kimura JH, Osdoby P, Caplan AI, Hascall VC: Electron microscopic and biochemical studies of proteoglycan polydispersity in chick limb bud chondrocyte cultures. J Biol Chem 253: 4721–4729, 1978.PubMedGoogle Scholar
  78. 78.
    Heinegård D, Lohmander S, Thyberg J: Cartilage proteoglycan aggregates. Electron-microscopic studies of native and fragmented molecules. Biochem J 175: 913–919, 1978.PubMedGoogle Scholar
  79. 79.
    Faltz LL, Reddi AH, Hascall GK, Martin D, Pita JC, Hascall VC: Characteristics of proteoglycans extracted from the Swarm rat chondrosarcoma with associative solvents. J Biol Chem 254: 1375–1380, 1979.PubMedGoogle Scholar

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© Martinus Nijhoff Publishers, Boston, The Hague, Dordrecht, Lancaster 1984

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  • C. Johan
  • O. Thyberg

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