Summary
A nitrous acid procedure has been shown to lead to the elimination of N-sulphates in sections of a series of tissues containing sulphated glycoconjugates. Two groups of sulphated glycoconjugate-containing tissues were used; one contained N-sulphates and other was devoid of such groupings. In the first group of tissues, mast cells of different origins and renal glomeruli in the rat were employed. Xiphoid and tracheal cartilage matrix, submandibular and sublingual gland acini and gastric, duodenal and colonic mucosae were used in the second group. Sections were treated with nitrous acid and then stained with Alcian Blue pH 1.0, high iron diamine or Aldehyde Fuchsin for sulphated glycoconjugates. Such treatment was found to diminish the staining intensities exclusively in N-sulphated glycoconjugate-containing structures such as mast cell granules and renal glomerular basement membrane, providing a means of chemically eliminating N-sulphates of glycoconjugates in tissues.
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References
Barrett, K. E. &Metcalfe, D. D. (1987) Heterogeneity of mast cells in the tissues of the respiratory tract and other organ systems.Am. Rev. Respir. Dis. 135, 1190–5.
Cifonelli, J. A. (1968) Reaction of heparitin sulfate with nitrous acid.Carbohydr. Res. 8, 233–42.
Dische, Z. &Borenfreund, E. (1950) A spectrophotometric method for the microdetermination of hexosamines.J. Biol. Chem. 184, 517–22.
Enerbäck, L. (1987) Mucosal mast cells in the rat and in man.Int. Archs. Allergy appl. Immun. 82, 249–55.
Enerbäck, L., Kolset, S. O., Kusche, M., Hjerpe, A. &Lindahl, U. (1985) Glycosaminoglycans in rat mucosal mast cells.Biochem. J. 227, 661–8.
Engel, U. R. (1971) Glycosaminoglycans in the aorta of six animal species.Atherosclerosis 13, 45–60.
Fischer, E. R. &Lillie, R. D. (1954) The effect of methylation on basophilia.J. Histochem. Cytochem. 2, 81–7.
Gilead, L., Livni, N., Eliakim, R., Ligumsky, M., Fich, A., Okon, E., Rachmilewitz, D. &Razin, E. (1987) Human gastric mucosal mast cells are chondroitin sulphate E-containing mast cells.Immunology 62, 23–8.
Horton, D. &Philips, K. D. (1973) The nitrous acid deamination of glycosides and acetates of 2-amino-2-deoxy-D-glucose.Carbohydr. Res. 30, 367–74.
Kanwar, Y. S. &Farquhar, M. G. (1979a) Presence of heparan sulfate in the glomerular basement membrane.Proc. Natl. Acad. Sci. USA 76, 1303–7.
Kanwar, Y. S. &Farquhar, M. G. (1979b) Isolation of glycosaminoglycans (heparan sulfate) from glomerular basement membranes.Proc. Natl. Acac. Sci. USA 76, 4493–7.
Lagunoff, D. &Warren, G. (1962) Determination of 2-deoxy-2-sulfoaminohexose content of mucopolysaccharides.Arch. Biochem. Biophys. 99, 396–400.
Lev, R. &Spicer, S. S. (1964) Specific staining of sulphate groups with alcian blue at low pH.J. Histochem. Cytochem. 12, 309.
Lillie, R. D. &Fullmer, H. M. (1976) Chemical end groups. InHistopathologic Techniques and Practical Histochemistry. 4th edn, pp. 217–326. New York: McGraw-Hill.
Lindahl, U., Bäckström, G., Jansson, L. &Hallen, A. (1973) Biosynthesis of heparin. II. Formation of sulfamino groups.J. Biol. Chem. 248, 7234–41.
Linker, A. &Hovingh, P. (1984) Structural studies on heparin. Tetrasaccharides obtained by heparinase degradation.Carbohydr. Res. 127, 75–94.
Moore, R. D. &Schoenberg, M. D. (1957) Low temperature sulfation of tissue and demonstration of metachromasy.Stain Technol. 32, 245–7.
Murata, K., Nakazawa, K. &Hamai, A. (1975) Distribution of acidic glycosaminoglycans in the intima, media and adventitia of bovine aorta and their anticoagulant properties.Atherosclerosis 21, 93–103.
Parmley, R. T., Eguchi, M., Spicer, S. S., Alvarez, C. J. &Austin, R. L. (1980) Ultrastructural cytochemistry and autoradiography of complex carbohydrates in heterophil granulocytes from rabbit bone marrow.J. Histochem. Cytochem. 28, 1067–80.
Pearse, A. G. E. (1985) Carbohydrates and mucosubstances. InHistochemistry: Theoretical and Applied. Vol 2:Analytical Technology, 4th edn, pp. 675–753. Edinburgh, London, Melbourne, New York: Churchill Livingstone.
Shively, J. E. &Conrad, H. E. (1970) Stoichiometry of the nitrous acid deaminative cleavage of model amino sugar glycosides and glycosaminoglycuronans.Biochemistry 9, 33–43.
Shively, J. E. &Conrad, H. E. (1976) Formation of anhydrosugars in the chemical depolymerization of heparin.Biochemistry 15, 3932–42.
Spicer, S. S. (1960) A correlative study of the histochemical properties of rodent acid mucopolysaccharides.J. Histochem. Cytochem. 8, 18–35.
Spicer, S. S. (1965) Diamine methods for differentiating mucosubstances histochemically.J. Histochem. Cytochem. 13, 211–34.
Spicer, S. S. &Lillie, R. D. (1959) Saponification as a means of selectively reversing the methylation blockade of tissue basophilia.J. Histochem. Cytochem. 7, 123–5.
Spicer, S. S., Horn, R. G. &Leppi, T. J. (1967) Histochemistry of connective tissue mucopolysaccharides. InConnective Tissue (Wagner, B. M. & Smith, D. E. eds), pp. 251–303. Baltimore: Williams & Wilkins.
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Hirabayashi, Y., Shimizu, S. & Yamada, K. A nitrous acid procedure as a selective histochemical means of eliminating the N-sulphates of glycoconjugates. Histochem J 21, 687–692 (1989). https://doi.org/10.1007/BF01002833
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DOI: https://doi.org/10.1007/BF01002833