Summary
The cytochemical localization of 5′-nucleotidase (5′-AMPase), and its validity, were investigated in parotid and submandibular acinar cells of a rat. Biochemical determinations showed that adequate treatment with glutaraldehyde could minimize the loss of enzymatic activity, and that 5′-AMPase and non-specific alkaline phosphatase (β-GPase) possessed different pH optima.
The cytochemical distribution of the reaction products from the 5′-AMPase activity was distinct from those of β-GPase. 5′-AMPase activity was localized on the surface membranes of acinar, ductal and myoepithelial cells of both salivary glands. β-GPase activity was evenly distributed on the entire plasma membranes of myoepithelial cells and on the basal plasmalemma of acinar cells. The reaction products, which appeared on the luminal and lateral plasma membranes of the acinar cells, were presumed to reflect the presence of 5′-AMPase, while those on the myoepithelial surface and basal plasma membranes of the acinar cells demonstrated both 5′-AMPase and β-GPase.
The results indicate that 5′-AMPase activity can be utilized as a reliable marker enzyme of plasma membranes in the salivary acinar cells.
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References
Ahmed, Z., Reis, J.L.: The activation and inhibition of 5′-nucleotidase. Biochem. J. 69, 386–387 (1958)
Bernstein, H.G., Weiss, J., Luppa, H.: Cytochemical investigations on the localization of 5′-nucleotidase in the rat hippocampus with special reference to synaptic regions. Histochemistry 55, 261–267 (1978)
Bodansky, O., Schwartz, M.K.: 5′-nucleotidase. In: Advance in clinical chemistry. (eds., O. Bodansky and C.P. Stewart) Vol. II pp. 277–328. New York: Academic Press 1968
Bogart, B.I.: The fine structural localization of alkaline and acid phosphatase activity in the rat submandibular gland. J. Histochem. Cytochem. 16, 572–581 (1968)
Borgers, M., Schaper, J., Schaper, W.: Localization of specific phosphatase activities in canine coronary blood vessels and heart muscle. J. Histochem. Cytochem. 19, 526–539 (1971)
Borgers, M., Thoné, F.: The inhibition of alkaline phosphatase by L-p-bromotetramisole. Histochemistry 44, 277–280 (1975)
El-Aaser, A.A., Reid, E.: Rat liver 5′-nucleotidase. Histochem. J. 1, 417–437 (1969)
El-Aaser, A.A., Fitzsimons, J.T.R., Hinton, R.H., Norris, K.A., Reid, E.: Electron microscopic examination of hepatic subcellular fractions for phosphatases. Histochem. J. 5, 199–223 (1973)
Essner, E., Novikoff, A.B., Masek, B.: Adenosine triphosphatase and 5-nucleotidase activities in the plasma membrane of liver cells as revealed by electron microscopy. J. Biophys. Biochem. Cytol. 4, 711–729 (1958)
Farquhar, M.G., Bergeron, J.J.M., Palade, G.E.: Cytochemistry of Golgi fractions prepared from rat liver. J. Cell Biol. 60, 8–25 (1974)
Garrett, J.R., Parsons, P.A.: Alkaline phosphatase and myoepithelial cells in the parotid gland of the rat. Histochem. J. 5, 463–471 (1973)
Gartner, L.P., Seibel, W., Hiatt, J.L., Provenza, D.V.: Electron microscopic localization of 5′-nucleotidase in the stratum intermedium and ameloblasts. Histochem. J. 10, 115–122 (1978)
Klaushofer, K., Böck, P., Müller, M.: Studies on 5′-nucleotidase histochemistry. I 5′-nucleotidase activity in the developing esophagus of the mouse. Histochemistry 39, 41–55 (1974)
Lowry, O.H., Rosenbrough, N.J., Farr, A.L., Randell, R.J.: Protein measurement with the folin phenol reagents. J. Biol. Chem. 193, 265–275 (1951)
Magnusson, B.C., Heyden, G., Svensson, S.E.: Histochemical observations on 5′-nucleotidase activity in squamous cell carcinomas of the rat. Histochemistry 42, 1–8 (1974)
Mayahara, H., Hirano, H., Saito, T., Ogawa, K.: The new lead citrate method for the ultracytochemical demonstration of activity of non-specific alkaline phosphatase (orthophosphoric monoester phosphohydrolase). Histochemistry 11, 88–96 (1967)
Morré, D.J., Ovtracht, L.: Dynamics of the Golgi apparatus: Membrane differentiation and membrane flow. Int. Rev. Cytol. [Suppl.] 5, 61–188 (1977)
Ray, T.: A modified method for the isolation of the plasma membrane from rat liver. Biochim. Biophys. Acta, 196, 1–9 (1970)
Reid, E.: Membrane system. In: Enzyme Cytology (ed. D.B. Roodyn) pp 321–406. London: Academic Press 1967
Rouser, G., Siakotos, A.N., Fleischer, S.: Quantitative analysis of phospholipids by the thin-layer chromatography and phosphorus analysis of spots. Lipids 1, 85–86 (1966)
Uusitalo, R.J., Karnovsky, M.J.: Surface localization of 5′-nucleotidase on the mouse lymphocytes. J. Histochem. Cytochem. 25, 87–96 (1977)
Wachstein, M., Meisel, E.: Histochemistry of hepatic phosphatase at a physiological pH. Am. J. Clin. Pathol. 27, 13–23 (1957)
Widnell, C.C.: Cytochemical localization of 5′-nucleotidase in subcellular fractions isolated from rat liver. I. Origin of 5′-nucleotidase activity in microsomes. J. Cell Biol. 52, 542–558 (1972)
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Yamashina, S., Kawai, K. Cytochemical studies on the localization of 5′-nucleotidase in the acinar cells of the rat salivary glands. Histochemistry 60, 255–263 (1979). https://doi.org/10.1007/BF00500655
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DOI: https://doi.org/10.1007/BF00500655