Summary
By means of a simultaneous azo coupling method N-acetyl-β-galactosaminidase has been detected preponderantly in the lysosomes of various organs (e.g. epididymis, spleen, kidney, liver, intestine and brain) from mice, guinea-pigs and rats.
The recommended incubation medium consists of 3.0–5.0 mg naphthol AS-BI N-acetylβ-galactosaminide (dissolved in ethylene glycol monomethyl ether) and 0.5–1.0 ml hexazonium-p-rosaniline in 10 ml 0.05 M citrate or citric acid-phosphate buffer, pH 4.0.
Parallely, N-acetyl-β-glucosaminidase has been demonstrated with the same assay replacing the naphthol AS-BI N-acetyl-β-galactosaminide by the corresponding glucosaminide.
In principal both enzymes displayed an identical distribution pattern and were inhibited to a similar extent by N-acetyl-β-glucosamine and N-acetyl-β-galactosamine or by the corresponding lactone. The activity of glucosaminidase, however, was always higher in comparison with β-galactosaminidase. Therefore some evidence occurs also from the histochemical point of view that β-galactosaminidase and β-glucosaminidase may represent one single lysosomal enzyme which preferentially attacks N-acetyl-β-glucosaminides.
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References
Barka, T., Anderson, P. J.: Histochemistry. New York-Evanston-London: Harper and Row, Inc. 1963.
Barman, Th. E.: Enzyme handbook, vol. II. Berlin-Heidelberg-New York: Springer 1969.
Bosmann, H. B., Bernacki, R. J.: Glycosidase activity. Glycosidase activity in L5178Y mouse leukemic cells and the activity of acid phosphatase, β-galactosidase and β-Nacetylgalactosaminidase and β-N-acetylglucosaminidase in a synchronous L5178Y cell population. Exp. Cell Ee-s. 61, 379–386 (1970).
Burstone, M. S.: Enzyme histochemistry and its application in the study of neoplasms. New York: Academic Press 1962.
Chang, J. P., Hori, S. H.: The section freeze-substitution technique: I. Method. J. Histochem. Cytochem. 9, 292–300 (1961).
Conchie, J., Hay, A. J., Strachan, I., Levvy, G. A.: Inhibition of glycosidases by aldonolactones of corresponding configuration. Preparation of (1→5)-lactones by catalytic oxidation of pyranoses and study of their inhibitory properties. Biochem. J. 102, 929–941 (1967).
Findlay, J., Levvy, G. A., Marsh, C. A.: Inhibition of glycosidases by aldonolactones of corresponding configuration. 2. Inhibitors of β-N-acetylglucosaminidase. Biochem. J. 69, 467–476 (1958).
Frohwein, Y. Z., Gatt, S.: Enzymatic hydrolysis of sphingolipids. VI. Hydrolysis of ceramide glycosides by calf brain β-N-acetylhexosaminidase. Biochemistry 6, 2783 (1967).
Gossrau, R.: Histochemische, fluoreszenzmikroskopische und experimentelle Untersuchungen am Reizleitungssystem von Goldhamster, Maus und Ratte. Histochemie 26, 44–60 (1971).
Gossrau, R.: Verwendung der Gefriertrocknung nach Lowry für histochemische Untersuchungen. (1972, in Vorbereitung.)
Gossrau, R.: Unpublished.
Hayashi, M.: Distribution of β-glucuronidase activity in rat tissues employing the naphthol AS-BI glucuronide hexazonium pararosanilin method. J. Histochem. Cytochem. 12, 659–669 (1964).
Hayashi, M.: Histochemical demonstration of N-acetyl-β-glucosaminidase employing naphthol AS-BI N-acetyl-β-glucosaminide as substrate. J. Histochem. Cytochem. 13, 355–360 (1965).
Holt, S. J.: Factors governing the validity of staining methods for enzymes, and their bearing upon the Gomori acid phosphatase technique. Exp. Cell Res., Suppl. 7, 1–27 (1959).
Lojda, Z.: Indigogenic methods for glycosidases. II. Improved method for β-D-galactosidase and its application to localization studies of the enzymes in the intestine and in other tissues. Histochemie 23, 266–288 (1970).
Lojda, Z.: Indigogenic methods for glycosidases. IV. An improved method for the demonstration of β-glucuronidase. Histochemie 27, 182–192 (1971b).
Lojda, Z.: Personal communication.
Lojda, Z., Kraml, J.: Indigogenic methods for glycosidases. III. An improved method with 4-Cl-5-Br-3-indolyl-β-D-fucoside and its application in studies of enzymes in the intestine, kidney, and other tissues. Histochemie 25, 195–207 (1971a).
Lojda, Z., Papoušek, F.: Basis of the histochemical demonstration of enzymes. Brno, Czechoslovac Society of Histo- and Cytochemistry, 1–108 (1970) [Czech.].
Lojda, Z., Ploeg, M. van der, Duijn, P. van: Phosphates of the naphthol As series in the quantitative determination of alkaline and acid phosphatase activities “in situ” studied in polyacrylamide membrane model systems and by cytospectrophotometry. Histochemie 11, 13–22 (1967).
Lojda, Z., Večerek, B., Pelichovă, H.: Some remarks concerning the histochemical detection of acid phosphatase by azo-coupling reactions. Histochemie 3, 428–454 (1964).
Lowry, O. H.: The quantitative histochemistry of the brain. Histological sampling. J. Histochem. Cytochem. 1, 420–428 (1953).
Mackenzie, A., Wilson, A. M., Dennis, P. F.: Further observations on histochemical changes in scrapie mouse brain. J. comp. Path. 78, 489–498 (1968).
Pearse, A. G. E.: Histochemistry. Theoretical and applied, 3rd ed., vol. I. London: Churchill 1968.
Pugh, D., Leaback, D. H., Walker, P. G.: Studies on glucosaminidase. N-acetyl-β-glucosaminidase in rat kidney. Biochem. J. 65, 464–469 (1957).
Robinson, D., Stirling, J. L.: N-acetyl-β-D-glucosaminidase in human spleen. Biochem. J. 107, 321–327 (1968).
Sato, K.: Histochemical demonstration of N-acetyl-β-galactosaminidase in the human tissues. Tohoku J. exp. Med. 97, 279–282 (1969).
Shuter, E. R., Robins, E., Freeman, M. L., Jungalwala, F. B.: β-Hexosaminidases in the nervous system: The quantitative histochemistry of β-glucosaminidase and β-galactosaminidase in the cerebellar cortex and subjacent white matter. J. Histochem. Cytochem. 18, 271–277 (1970).
Walker, J. W., Woollen, P. G., Heyworth, R.: Studies on glucosaminidase. 5. Kidney N-acetyl-β-galactosaminidase. Biochem. J. 79, 288–294 (1961).
Weissmann, B., Hadjiioannou, S., Tornheim, J.: Oligosaccharase activity of β-N-acetyl-D-glucosaminidase of beef liver. J. biol. Chem. 239, 59–63 (1964).
Winckler, J.: Zum Einfrieren von Gewebe in Stickstoff-gekühltem Propan. Histochemie 23, 44–50 (1970a).
Winckler, J.: Verwendung gefriergetrockneter Kyrostatschnitte für histologische und histochemische Untersuchungen. Histochemie 24, 168–186 (1970b).
Woollen, J. W., Walker, P. G., Heyworth, R.: Studies on glucosaminidase. 6. N-acetyl-β-glucosaminidase and N-acetyl-β-galactosaminidase activities of a variety of enzyme preparations. Biochem. J. 79, 294–298 (1961).
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Gossrau, R. On the histochemical demonstration of N-acetyl-β-galactosaminidase. Histochemie 29, 315–324 (1972). https://doi.org/10.1007/BF00279814
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DOI: https://doi.org/10.1007/BF00279814