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An ultrastructural study of histochemical staining of complex carbohydrates in the mouse posterior vitreous body

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Summary

The vitreous body contains complex carbohydrates that can be demonstrated morphologically. Vitreous hyaluronic acid is very soluble but it can be precipitated by cetylpyridinium chloride (CPC) while being cross-linked by glutaraldehyde. Oligosaccharide chains of vitreous glycoproteins are fixed with glutaraldehyde alone. Mouse eyes were fixed with glutaraldehyde or glutaraldehyde and CPC and the complex carbohydrates of the posterior vitreous cortex were studied by electron microscopy. Cationic dyes were used in the fixative or for block-staining on most fixed tissue blocks to allow detailed observations of complex carbohydrates. Most blocks were postfixed with OsO4. The hyaluronic-acid domain on vitreous collagen fibrils sequentially contracted and expanded in size with various histochemical manipulations. Contraction of the domain of hyaluronic acid generally indicates an increased charge density. OsO4 contributes considerable charge density upon forming osmate esters, but tissue postfixed with OsO4 contained large globular forms of hyaluronic acid rather than the small globules observed in non-osmicated preparations. A model is proposed to explain the seemingly paradoxical findings by reference to suggested mechanisms of polysasaccharideligand-OsO4 interactions.

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References

  • Allen DE, Perrin DD (1979) Electron cytochemical stains based on metal chelation. Int Rev Cytol 61:63–84

    Google Scholar 

  • Allen WS, Otterbein EC, Wardi AH (1977) Isolation and characterization of the sulfated glycosaminoglycans of the vitreous body. Biochim Biophys Acta 498:167–175

    Google Scholar 

  • Ashton N, Tripathi R (1972) The argyrophilic mosaic of the internal limiting membrane of the retina. Exp Eye Res 14:49–52

    Google Scholar 

  • Balazs EA (1958) Physical chemistry of hyaluronic acid. Fed Proc 17:1086–1093

    Google Scholar 

  • Balazs EA (1961) Molecular morphology of the vitreous body. In: Smelser GK (ed) The structure of the eye. Academic Press, New York, pp 293–310

    Google Scholar 

  • Balazs EA (1982) Functional anatomy of the vitreus. In: Jakobiec FA (ed) Ocular anatomy, embryology, and teratology. Harper and Row, Philadelphia, pp 425–440

    Google Scholar 

  • Behnke O, Zelander T (1970) Preservation of intercellular substances by the cationic dye Alcian blue in preparative procedures for electron microscopy. J Ultrastruct Res 31:424–438

    Google Scholar 

  • Chakrabarti B, Hultsch E (1976) Owl monkey vitreous: a novel model for hyaluronic acid structural studies. Biochem Biophys Res Commun 71:1189–1193

    Google Scholar 

  • Chen CH, Chen SC (1981) Studies on soluble proteins of vitreous in experimental animals. Exp Eye Res 32:381–388

    Google Scholar 

  • Dische Z, Danilczenko A, Zelmenis G (1958) The neutral heteropolysaccharides in connective tissue. In: Wolstenholme GEW, O'Connor (eds) Ciba foundation symposium on the chemistry and biology of mucopolysaccharides. Little, Brown and Co, Boston, pp 116–139

    Google Scholar 

  • Eisenstein R, Sorgente N, Kuettner KE (1971) Organization of extracellular matrix in epiphyseal growth plate. Am J Pathol 65:515–534

    Google Scholar 

  • Fessler JH, Fessler LI (1966) Electron microscopic visualization of the polysaccharide hyaluronic acid. Proc Natl Acad Sci USA 56:141–147

    Google Scholar 

  • Fine BS, Tousimis AJ (1961) The structures of the vitreous body and the suspensory ligaments of the lens. Arch Ophthalmol 65:95–110

    Google Scholar 

  • Forrester JV, Grierson I, Lee WR (1980) Vitreous membrane formation after experimental vitreous haemorrhage. Albrecht Von Graefes Arch Klin Exp Ophthalmol 212:227–242

    Google Scholar 

  • Forrester JV, Lackie JM (1981) Effect of hyaluronic acid on neutrophil adhesion. J Cell Sci 50:329–344

    Google Scholar 

  • François J, Victoria-Troncoso V, Albarran E (1970) The histochemical structure of the vitreous fibers studied by phase contrast microscopy. Am J Ophthalmol 69:763–773

    Google Scholar 

  • Hadler NM, Dourmashkin RR, Nermut MV, Williams LD (1982) Ultrastructure of a hyaluronic acid matrix. Proc Natl Acad Sci USA 79:307–309

    Google Scholar 

  • Hascall VC, Hascall GK (1981) Proteoglycans. In: Hay ED (ed) Cell biology of extracellular matrix. Plenum Press, New York, pp 39–63

    Google Scholar 

  • Hofmann H, Schmut O (1978) Die unterschiedliche Wirkung von Fe(II)-und Fe(III)-lonen auf die Hyaluronsäure des Glaskörpers. Klin Monatsbl Augenheilkd 172:261–263

    Google Scholar 

  • Hultsch E (1981) The vitreous of the baby owl monkey: a model for rapid and complete gel-liquefaction. Dev Ophthalmol 2:1–7

    Google Scholar 

  • Kanwar YS, Farquhar MG (1979) Presence of heparan sulfate in the glomerular basement membrane. Proc Natl Acad Sci USA 76:1303–1307

    Google Scholar 

  • Kelly JW, Bloom GD, Scott JE (1963) Quaternary ammonium compounds in connective tissue histochemistry: I. Selective unblocking. J Histochem Cytochem 11:791–798

    Google Scholar 

  • Lam KW, Ashrafzadeh MT, Lee CB (1972) Vitreous membranes: induction in rabbits by intravitreous leukocyte injections. Arch Ophthalmol 88:655–658

    Google Scholar 

  • Laurent TC (1970) Structure of hyaluronic acid. In: Balazs EA (ed) Chemistry and molecular biology of the intercellular matrix, vol 2. Academic Press, New York, pp 703–732

    Google Scholar 

  • Luft JH (1971) Ruthenium red and violet. II. Fine structural localization in animal tissues. Anat Rec 171:369–415

    Google Scholar 

  • Luft JH (1976) The structure and properties of the cell surface coat. Int Rev Cytol 45:291–382

    Google Scholar 

  • Markwald RR, Fitzharris TP, Bernanke DH (1979) Morphologic recognition of complex carbohydrates in embryonic cardiac extracellular matrix. J Histochem Cytochem 27: 1171–1173

    Google Scholar 

  • Orkin RW, Toole BP (1980) Isolation and characterization of hyaluronidase from cultures of chick embryo skin-and muscle-derived fibroblasts. J Biol Chem 255:1036–1042

    Google Scholar 

  • Österlin SE, Balazs EA (1968) Macromolecular composition and fine structure of the vitreous in the owl monkey. Exp Eye Res 7:534–545

    Google Scholar 

  • Pfeffer BA, Hassell JR, Newsome DA (1981) Composition and interaction of components of monkey vitreous. Invest Ophthalmol Vis Sci 20 (Suppl):201

    Google Scholar 

  • Phillips GO (1970) Interaction between glycosaminoglycans and organic cations. In: Balazs EA (ed) Chemistry and molecular biology of the intercellular matrix, vol 2. Academic Press, New York, pp 1033–1065

    Google Scholar 

  • Poole AR, Pidoux I, Reiner A, Rosenberg L (1982) An immunoelectron microscopes study of the organization of proteoglycan monomer, link protein, and collagen in the matrix of articular cartilage. J Cell Biol 93:921–937

    Google Scholar 

  • Quintarelli G, Vocaturo A, Bellocci M, Rodén L, Ippolito E, Baker JR (1974) Preliminary ultrastructural demonstration of hyaluronic acid-proteoglycan interaction in cartilage matrix. Am J Anat 140:433–438

    Google Scholar 

  • Rentsch FJ (1976) The fine structure of the normal and pathological vitreous body as revealed by ruthenium-red. In: Yamada E, Mishima S (eds) The structure of the eye, vol 3. Japanese Journal of Ophthalmology, Tokyo, pp 19–37

  • Resch J, Tunkel D, Stoeckert C, Beer M (1980) Osmium-labeled polysaccharides for atomic microscopy. J Mol Biol 138:673–683

    Google Scholar 

  • Rhodes RH (1978) Development of the human optic disc: light microscopy. Am J Anat 153:601–615

    Google Scholar 

  • Rhodes RH (1982a) Localization of fucosyl glycoproteins in the vitreous body and zonules by lectin histochemistry. Invest Ophthalmol Vis Sci 22 (Suppl):4

    Google Scholar 

  • Rhodes RH (1982b) An ultrastructural study of the complex carbohydrates of the mouse posterior vitreoretinal juncture. Invest Ophthalmol Vis Sci 22:460–477

    Google Scholar 

  • Rhodes RH (1983) Ultrastructure of complex carbohydrates in the mouse posterior chamber and vitreous base. Invest Ophthalmol Vis Sci 24 (Suppl):262

    Google Scholar 

  • Rhodes RH, Mandelbaum SH, Minckler DS, Cleary PE (1982) Tritiated fucose incorporation in the vitreous body, lens and zonules of the pigmented rabbit. Exp Eye Res 34:921–931

    Google Scholar 

  • Riemersma JC (1963) Osmium tetroxide fixation of lipids: nature of the reaction products. J Histochem Cytochem 11:436–442

    Google Scholar 

  • Ruggeri A, Dell'Orbo C, Quacci D (1975) Electron microscopic visualization of proteoglycans with Alcian Blue. Histochem J 7:187–197

    Google Scholar 

  • Sanders D, Peyman GA, Fishman G, Vlchek J, Korey M (1975) The toxicity of intravitreal whole blood and hemoglobin. Albrecht Von Graefes Arch Klin Exp Ophthalmol 197:255–267

    Google Scholar 

  • Sannes PL, Katsuyama T, Spicer SS (1978) Tannic acid-metal salt sequences for light and electron microscopic localization of complex carbohydrates. J Histochem Cytochem 26:55–61

    Google Scholar 

  • Schofield BH, Williams BR, Doty SB (1975) Alcian Blue staining of cartilage for electron microscopy. Histochem J 7:139–149

    Google Scholar 

  • Schwarz W (1961) Electron microscopic observations of the human vitreous body. In: Smelser GK (ed) The structure of the eye. Academic Press, New York, pp 283–291

    Google Scholar 

  • Schwarz W (1976) Electron microscopic study on the gel of the central part of the corpus vitreum in the ox. Cell Tissue Res 168:271–275

    Google Scholar 

  • Scott JE (1955) The solubility of cetylpyridinium complexes of biological polyanions in solution of salts. Biochim Biophys Acta 18:428–429

    Google Scholar 

  • Scott JE (1960) Aliphatic ammonium salts in the assay of acidic polysaccharides from tissues. Methods Biochem Anal 8:145–197

    Google Scholar 

  • Scott JE, Dorling J (1965) Differential staining of acid glycosaminoglycans (mucopolysaccharides) by Alcian Blue in salt solutions. Histochemie 5:221–233

    Google Scholar 

  • Scott JE, Stockwell RA (1967) On the use and abuse of the critical electrolyte concentration approach to the localization of tissue polyanions. Letter-to-the-editor. J Histochem Cytochem 15:111–113

    Google Scholar 

  • Shepard N, Mitchell N (1976) Simultaneous localization of proteoglycan by light and electron microscopy using toluidine blue O. A study of epiphyseal cartilage. J Histochem Cytochem 24:621–629

    Google Scholar 

  • Smith JW, Serafini-Fracassini A (1967) The relationship of hyaluronate and collagen in the bovine vitreous body. J Anat 101:99–112

    Google Scholar 

  • Snowden JM, Swann DA (1980) Vitreous structure. V. The morphology and thermal stability of vitreous collagen fibers and comparison to articular cartilage (type II) collagen. Invest Ophthalmol Vis Sci 19:610–618

    Google Scholar 

  • Snowden JM, Eyre DR, Swann DA (1982) Vitreous structure. VI. Age-related changes in the thermal stability and crosslinks of vitreous, articular cartilage and tendon collagens. Biochim Biophys Acta 706:153–157

    Google Scholar 

  • Sorvari TE, Näntö V (1971) Investigations of dye binding in the sequential staining of mucopolysaccharides by Alcian Yellow-Alcian Blue. II. Spot tests and experiments on dye-mucopolysaccharide (glycosaminoglycan) precipitates. Histochem J 3:269–277

    Google Scholar 

  • Stainer GA, Peyman GA, Berkowitz R, Tessler HH (1976) Intraocular lysozyme in experimental uveitis in rabbits: aqueous and vitreous assay. Invest Ophthalmol 15:312–315

    Google Scholar 

  • Swann DA (1980) Chemistry and biology of the vitreous body. Int Rev Exp Pathol 22:2–64

    Google Scholar 

  • Swann DA, Sotman S (1980a) Non-collagenous structural proteins in the vitreous. Invest Ophthalmol Vis Sci 19 (Suppl):145

    Google Scholar 

  • Swann DA, Sotman SS (1980b) The chemical composition of bovine vitreous-humour collagen fibres. Biochem J 185:545–554

    Google Scholar 

  • Swann DA, Sotman SL, Hermann H, Garg HG (1981) Isolation and partial characterization of a proteochondroitin sulfate and a proteoheparan sulfate from the bovine vitreous. Invest Ophthalmol Vis Sci 20 (Suppl):139

    Google Scholar 

  • Szirmai JA (1963) Quantitative approaches in the histochemistry of mucopolysaccharides. J Histochem Cytochem 11:24–34

    Google Scholar 

  • Takagi M, Parmley RT, Toda Y, Austin RL (1982) Ultrastructural cytochemistry and immunocytochemistry of sulfated glycosaminoglycans in epiphyseal cartilage. J Histochem Cytochem 30:1179–1185

    Google Scholar 

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  1. Department of Pathology, School of Medicine, University of Southern California, 2011 Zonal Avenue, 90033, Los Angeles, CA, USA

    R. H. Rhodes

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Supported in part by American Cancer Society Institutional Research Grant IN-85-M-15, University of Tennessees and NIH Grants EY-03828 and EY-04705

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Rhodes, R.H. An ultrastructural study of histochemical staining of complex carbohydrates in the mouse posterior vitreous body. Histochemistry 78, 125–143 (1983). https://doi.org/10.1007/BF00491119

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  • DOI: https://doi.org/10.1007/BF00491119

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