Summary
Five new methods, based upon the selective oxidation of sialic acid residues with 0.4mm periodic acid in approximately 1m hydrochloric acid at 4°C for 1 h (PA*), have been devised for the simultaneous visualization of neutral sugars and either sialic acid and its side chainO-acyl variants orO-sulphate ester. In the first of these, the selective periodate oxidation—borohydride reduction—saponification—selective periodate oxidation—Thionin Schiff—saponification—borohydride reduction—periodic acid—Schiff (PA*—Bh—KOH—PA*—T—KOH—Bh—PAS) technique, sialic acids withO-acyl substituents at C7, C8 or C9 (or which have two of three side chainO-acyl substituents) stain blue while neutral sugars with periodate-sensitivevicinal diols (hexose, 6-deoxyhexose, andN-acetylhexosamine) stain magenta. The second method, the saponification—selective periodate oxidation—Thionin Schiff—saponification—borohydride reduction—periodic acid—Schiff (KOH—PA*—T—KOH—Bh—PAS), stains all sialic acids blue and neutral sugars magenta. In the third procedure, the selective periodate oxidation—Thionin Schiff—borohydride reduction—periodic acid—Schiff—saponification (PA*—T—Bh—PAS—KOH) method, sialic acids without side chain substituents (or which have anO-acyl substituent at C7) stain blue and neutral sugars stain magenta. In the fourth method, the saponification-selective periodate oxidation—borohydride reduction—Alcian Blue pH 1.0—periodic acid—Schiff (KOH—PA*—Bh—AB1.0—PAS) technique,O-sulphate esters stain aquamarine blue and neutral sugars stain magenta. In all of these techniques mixtures of the components stain in various shades of purple. Performance of the KOH—PA*—Bh—AB1.0—PAS technique without the Alcian Blue pH 1.0 step provides a method for the selective identification of neutral sugars in macromolecules that also contain sialic acids.
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References
BARGER, J. D. & DELAMATER, E. D. (1948) The use of thionyl chloride in the preparation of Schiff reagent.Science 108, 121–2.
BOLAND, C. R., MONTGOMERY, C. K. & KIM, Y. S. (1982a) Alteration in human colonic mucin occurring with cellular differentiation and malignant transformation.Proc. Natl. Acad. Sci. USA 79, 2051–5.
BOLAND, C. R., MONTGOMERY, C. K. & KIM, Y. S. (1982b) A cancer associated mucin. Alteration in benign colonic polyps.Gastroenterology 82, 664–72.
BOLAND, C. R., LANCE, P., LEWIN, B., RIDDELL, R. H. & KIM, Y. S. (1984) Abnormal goblet cells glycoconjugates in rectal biopsies associated with an increased risk of neoplasia in patients with ulcerative colitis: early results of a prospective study.Gut 25, 1364–71.
BRESALIER, R. S., BOLAND, R. & KIM, Y. S. (1984) Characteristics of colorectal carcinoma cells with high metastatic potential.Gastroenterology 87, 115–22.
CHERESH, D. A. & REISFELD, R. A. (1984)O-acylation of disialo ganglioside GD3 by human melanoma cells creates a unique antigenic determinant.Science 225, 844–6.
COOPER, H. S. (1982) Peanut lectin binding sites in large bowel carcinoma.Lab. Invest. 47, 383–9.
CULLING, C. F. A., REID, P. E. & DUNN, W. L. (1977) A new histochemical method of use in the interpretation and diagnosis of adenocarcinoma and villous lesions in the large intestine.J. Clin. Path. 30, 1056–62.
CULLING, C. F. A., REID, P. E. & DUNN, W. L. (1979) A histochemical comparison of theO-acylated sialic acids of the epithelial mucins in ulcerative colitis, Crohn's disease and normal controls.J. Clin. Path. 32, 1272–7.
CULLING, C. F. A., REID, P. E. & WORTH, A. J. (1981a) Carbohydrate markers in the lower gastrointestinal tract, inMethods and Achievements in Experimental Pathology Cell Markers Vol 10 (edited by JASMIN, G. and CATIN, M.) pp. 77–100. Basel: S. Karger.
CULLING, C. F. A., REID, P. E., DUNN, W. L. & FREEMAN, H. J. (1981b) The relevance of the histochemistry of colonic mucins based upon their PAS reactivity.Histochem. J. 13, 899–903.
DAWSON, P. A., PATEL, J. & FILIPE, M. I. (1978) Variations in sialomucins in the mucosa of large intestine in malignancy: a quantitative and statistical analysis.Histochem. J. 10, 599–72.
DIAZ, S. & VARKI, A. (1985) Metabolic labelling of sialic acids in tissue culture cell lines: methods to identify substituted and modified radioactive neuraminic acids.Anal. Biochem. 150, 32–46.
EHSANULLAH, M., FILIPE, M. I. & GAZZARD, B. (1982a) Morphological and mucous secretion criteria for differential diagnosis of solitary ulcer syndrome and non-specific proctitis.J. Clin. Path. 35, 26–30.
EHSANULLAH, M., FILIPE, M. I. & GAZZARD, B. (1982b) Mucin secretion in inflammatory bowel disease: correlation with disease activity and dysplasia.Gut 23, 485–9.
FAKAN, F. & ADAMOCOVA, J. (1981) Diagnostic value of the PB—KOH—PAS method for identification of primary cancer from metastases.Acta. Histochem. 68, 176–80.
FENGER, C. & FILIPE, M. I. (1981) Mucin histochemistry of the anal canal epithelium. Studies of normal anal mucosa and mucosa adjacent to carcinoma.Histochem. J. 13, 921–30.
FILIPE, M. I. (1979) Mucins in the human gastrointestinal epithelium: a review.Invest. Cell Pathol. 2, 195–216.
FILIPE, M. I. (1984) Transitional mucosa.Histopathology 8, 707.
FILIPE, M. I. & BRANFOOT, A. C. (1976) Mucin histochemistry of the colon.Curr. Top. Path. 63, 143–78.
FILIPE, M. I., MUGHAL, S. & BUSSEY, H. J. (1980) Pattern of mucus secretion in the colonic epithelium in familial polyposis coli.Invest. Cell Path. 3, 329–43.
FRANZIN, G., SCAPA, A., DINA, R. & NOVELLI, P. (1981) ‘Transitional’ and hyperplastic—metaplastic mucosa occurring in solitary ulcer of the rectum.Histopathology 5, 27–33.
FRANZIN, G., GRIGIONI, W. F., DINA, R., SCARPA, A. & ZAMBONI, G. (1983a) Mucin secretion and morphological changes of the mucosa in non-neoplastic diseases of the colon.Histopathology 7, 707–18.
FRANZIN, G., ZAMBONI, G., DINA, R., SCARPA, A. & FRATTON, A. (1983b) Juvenile and inflammatory polyps of the colon — a histological histochemical study.Histopathology 7, 719–28.
FRANZIN, G., DINA, R., ZAMBONI, G., IANNUCCI, A., SCARPA, A. & NOVELLI, P. (1984) Hypoplastic (metaplastic) polyps of the colon.Am. J. Surg. Pathol. 8, 687–98.
GREAVES, P., FILIPE, M. I. & BRANFOOT, A. C. (1980) Transitional mucosa and survival in human colorectal cancer.Cancer 46, 764–70.
GREAVES, P., FILIPE, M. I., ABBAS, S. & ORMEROD, M. G. (1984) Sialomucins and carcinoembryonic antigen in the evolution of colorectal cancer.Histopathology 8, 825–34.
HABIB, N., SALEM, R., LUCK, R. J., BLOUNT, M. A., RIFAAT, M. A. & WOOD, C. B. (1984) A histochemical method that predicts local recurrence after curative resection in carcinoma of the colon and rectum.Surg. Gynecol. Obstet. 159, 436–8.
HAVERKAMP, J., SCHAUER, R., WEMBER, M., KAMBERLING, J. F. & VLIEGENTHART, J. F. P. (1975) Synthesis of 9-O-acetyl and 4,9-di-O-acetylneuraminic acid methylglycoside.Z. Physiol. Chem. 356, 1575–83.
ISAACSON, P. & ATTWOOD, P. R. A. (1979) Failure to demonstrate specificity of the morphological and histochemical changes in mucosa adjacent to colonic carcinoma (transitional mucosa).J. Clin. Pathol. 32, 214–18.
JASS, J. R., FILIPE, M. I., ABBAS, S., FALCON, C. A., WILSON, Y. & LOVELL, D. (1984) A morphologic and histochemical study of metaplastic polyps of the colorectum.Cancer 53, 510–15.
LEV, R., LANCE, P. & COMAVA, P. (1985) Histochemical and morphological studies of mucosa bordering rectosigmoid carcinoma. Comparison with normal, diseased and malignant epithelium.Human Pathol. 16, 151–61.
LILLIE, R. D. & PIZZOLATO, P. (1972) Histochemical use of borohydrides as aldehyde blocking reagents.Stain Technol. 47, 13–16.
LISTINSKI, C. M. & RIDDELL, R. H. (1981) Patterns of mucin secretion in neoplastic and non-neoplastic disease of the colon.Human Pathol. 12, 923–9.
MARCHEGGIANO, A., TANNONI, C., PALLONE, F., FRIESI, G., GALLUCI, M. & CAPRILLI, R. (1984) Abnormal patterns of mucin secretion after ureterosigmoidostomy.Human Pathol. 15, 647–50.
MONTERO, C. & SEGURA, D. I. (1980) Retrospective histochemical study of mucosubstances in adenocarcinomas of the gastrointestinal tract.Histopathology 4, 281–91.
MOWRY, R. W. (1963) Special value of methods that colour both acidic and vicinal hydroxyl groups in the histochemical study of mucins. With revised directions for the colloidal iron stain, the use of Alcian Blue 8GX and their combination with the periodic acid—Schiff reaction.Ann. NY Acad. Sci. 106, 402–23.
PARK, C. M., REID, P. E., WALKER, D. C. & MACPHERSON, B. R. (1987) A simple practical ‘Swiss roll’ method of preparing tissues for paraffin and methacrylate embedding.J. Microsc. in press.
REID, P. E. & CULLING, C. F. A. (1980) The periodate oxidation of carbohydrates in relation to the PAS reaction.J. Histotechnol. 3, 82–90.
REID, P. E., CULLING, C. F. A. & DUNN, W. L. (1973) Saponification induced increase in the periodic acid—Schiff reaction in the gastrointestinal tract mechanism and distribution of the reactive substance.J. Histochem. Cytochem. 21, 473–82.
REID, P. E., CULLING, C. F. A., RAMEY, C. W., DUNN, W. L. & CLAY, M. G. (1977) A simple method for the determination of theO-acetyl substitution pattern of the sialic acids of colonic epithelial glycoprotein.Can. J. Biochem. 55, 493–503.
REID, P. E., CULLING, C. F. A. & DUNN, W. L. (1978) A histochemical method for the demonstration of 9-O-acyl sialic acids. An investigation of bovine submaxillary mucin and intestinal mucins.J. Histochem. Cytochem. 26 187–92.
REID, P. E., CULLING, C. F. A., DUNN, W. L., RAMEY, C. W. & CLAY, M. G. (1984a) Chemical and histochemical studies of normal and diseased gastrointestinal tract. I. A comparison between histologically normal colon, colonic tumours, ulcerative colitis and diverticular disease of the colon.Histochem. J. 16, 235–51.
REID, P. E., DUNN, W. L., RAMEY, C. W., CORET, E., TRUEMAN, L. & CLAY, M. G. (1984b) Histochemical identification of side chain substitutedO-acylated sialic acids. The PAT—KOH—Bh—PAS and the PAPT—KOH—Bh—PAS procedures.Histochem. J. 16, 623–39.
REID, P. E., DUNN, W. L., RAMEY, C. W., CORET, E., TRUEMAN, L. & CLAY, M. G. (1984c) Histochemical studies of the mechanism of the periodic acid—phenyl-hydrazine—Schiff (PAPS) procedure.Histochem. J. 16, 641–9.
REID, P. E., OWEN, D. A., DUNN, W. L., RAMEY, C. W., LAZOSKY, D. A. & CLAY, M. G. (1985a) Chemical and histochemical studies of normal and diseased gastrointestinal tract. III. Changes in the histochemical and chemical properties of the epithelial glycoproteins in the mucosa close to colonic tumours.Histochem. J. 17, 171–82.
REID, P. E., OWEN, D. A., RAMEY, C. W., DUNN, W. L., JONES, E. A., LAZOSKY, D. A., ALLEN, E. (née Atkins), PARK, C. M. & CLAY, M. G. (1985b). Chemical and histochemical studies of normal and diseased human gastrointestinal tract. V. A differential diagnostic method for the histochemical classification of glycoproteins.Histochem. J. 17, 891–903.
REID, P. E., OWEN, D. A., RAMEY, C. W., DUNN, W. L., CLAY, M. G. & JONES, E. A. (1985c) Histochemical procedures for the simultaneous visualization of sialic acid, its side chainO-acyl variants andO-sulphate ester.Histochem. J. 17, 113–17.
RHATIGAN, R. M. & SAFFROS, R. O. (1979) Mucosal hyperplasia in colonic diverticula.Histopathology 3, 153–60.
SCHAUER, R. (1982) Chemistry metabolism and biological functions of sialic acid.Adv. Carb. Chem. Biochem. 40, 131–234.
SHUKLA, A. K. & SCHAUER, R. (1982). Fluorimetric determination of unsubstituted and 9 (8)-O-acylated sialic acids in erythrocyte membrane.Z. Physiol. Chem. 363, 255–62.
SCHULTE, B. A. & SPICER, S. S. (1983) Light microscopic histochemical detection of sugar residues in secretory glycoproteins of rodent and human tracheal glands with lectin—horseradish peroxidase conjugates and the galactose oxidase—Schiff sequence.J. Histochem. Cytochem. 31, 391–403.
SPICER, S. S. (1965) Diamine methods for differentiating mucosubstances histochemically.J. Histochem. Cytochem. 13, 211–34.
SUNTER, J. P., HIGGS, M. J. & COWAN, W. K. (1985) Mucosal abnormalities at the anastomosis site in patients who have had intestinal resection for colonic cancer.J. Clin. Pathol. 38, 385–9.
VAN DUIJN, P. (1956) A histochemical specific thionine—SO2 reagent and its use in a bi-colour method for deoxyribonucleic acid and periodic acid—Schiff positive substance.J. Histochem. Cytochem. 4, 55–63.
VARKI, A. & DIAZ, S. (1984) The release and purification of sialic acids from glycoconjugates: methods to minimize the loss and migration ofO-acetyl groups.Anal. Biochem. 137, 236–47.
VEH, R. W., CORFIELD, A. P. SCHAUER, R. & ANDREWS, K. H. (1979) The bovine submandibular gland. II. Histochemical and biochemical results. In Proceedings of the 5th International Symposium on Glycoconjugates, pp. 652–3. Thieme: Stuttgart.
VOLZ, D., REID, P. E., PARK, C. M., OWEN, D. A., DUNN, W. L. & RAMEY, C. W. (1986) Can ‘mild’ periodate oxidation be used for the specific histochemical identification of sialic acid residues?Histochem. J. 18, 579–82.
VOLZ, D., REID, P. E., PARK, C. M., OWEN, D. A. & Dunn, W. L. (1987) A new method for the selective periodate oxidation of total tissue sialic acids.Histochem. J., in press.
YONEZAWA, S., NAKAMURA, T., TANAKA, S. & SATO, E. (1982) Glycoconjugate withUlex europaeus agglutinin-1 binding sites in normal mucosa. Adenoma and carcinoma of the human large bowel.J. Nat. Can. Inst. 69, 777–83.
YONEZAWA, S., NAKAMURA, T., TANAKA, S., MARATA, K., NISHI, M. & SATO, E. (1983) Binding ofUlex europaeus agglutinin-1 in polyposis coli. Comparative study with solitary adenoma in the sigmoid colon and rectum.J. Nat. Can. Inst. 71, 17–24.
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Volz, D., Reid, P.E., Park, C.M. et al. Histochemical procedures for the simultaneous visualization of neutral sugars and either sialic acid and its side chainO-acyl variants orO-sulphate ester. I. Methods based upon the selective periodate oxidation of sialic acids. Histochem J 19, 249–256 (1987). https://doi.org/10.1007/BF01675683
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DOI: https://doi.org/10.1007/BF01675683