Summary
Specimens of the human parotid gland were studied by immuno-electron microscopy for the presence of amylase. Both the protein A-gold technique and the biotin-avidin-gold technique were used on the same specimens. Different fixations were tried. Amylase was detected in the zymogen granules in high amounts. This enzyme could even be seen in glutaraldehyde fixed and routinely embedded material. The subcellular localization of this enzyme opens a new field of functional morphological studies and studies in special tumours including acinic cell carcinomas.
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References
Amsterdam A, Schramm M (1966) Rapid release of the zymogen granule protein by osmium tetroxide and its retention during fixation by glutaraldehyde. J Cell Biol 29:199–207
Baskin DG, Erlandsen SL, Parsons JA (1979) Influence of hydrogen peroxide or alcoholic sodium hydroxide on the immunocytochemical detection of growth hormone and prolactin after osmium fixation. J Histochem Cytochem 27:1290–1292
Batsakis JG, Wozniak KJ, Regezi JA (1977) Acinous cell carcinoma: a histogenetic hypothesis. J Oral Surg 35:904–906
Bayer EA, Skutelsky E, Wilchek M (1979) The avidin-biotin-complex in affinity cytochemistry. Methods Enzymol 62:308–315
Bendayan M (1982) Double immunocytochemical labeling applying the protein A-gold technique. J Histochem Cytochem 30:81–85
Bendayan M, Roth J, Pardet A, Orci L (1980) Quantitative immunocytochemical localization of pancreatic secretory protein in subcellular compartments of the rat acinar cell. J Histochem Cytochem 28:149–160
Caselitz J, Seifert G, Grenner G, Schmidtberger R (1983) Amylase as an additional marker of salivary gland neoplasms. Pathol Res Pract 176:276–283
De Mey J (1983) Colloidal gold probes in immunocytochemistry. In: Polak JM, Van Noorden S (eds) Immunocytochemistry. John Wright & Sons Ltd, Bristol, London, Boston, pp 82–112
Eldred WD, Zucker C, Karten HJ, Yazulla S (1983) Comparison of fixation and penetration enhancement techniques for use in ultrastructural immunocytochemistry. J. Histochem Cytochem 31:285–292
Erlandson RA (1972) Ultrastructure of acinic cell carcinoma of the parotid gland. Arch Pathol 93:130–140
Erlandson RA (1981) Diagnostic transmission electron microscopy of human tumors. Masson Publishing, New York, pp 44–58
Faulk WP, Taylor GM (1971) An immunocolloid method for the electron microscopy. Immunochemistry 8:1081–1083
Geuze HJ, Slot JW, Von der Ley PA, Scheffer RCT, Griffith JM (1981) Use of colloidal gold particles in double labeling immunoelectron microscopy of ultrathin frozen tissue sections. J Cell Biol 89:653–665
Ghadially FN (1980) Diagnostic electron microscopy of tumours. Butterworths, London, pp 215–219
Heitzmann H, Richards FM (1974) Use of the avidin-biotin complex for specific staining of biological membranes in electron microscopy. Proc Natl Acad Sci USA 71:3537–3541
Hirtzler R, Oberman B, Kuliš M, Ljubešić N (1969) Azinuszelladenocarcinome der Speicheldrüsen. Arch Ohr Nas Kehlk Heilk 195:68–80
Hsu SM, Raine L, Fanger H (1981) Use of Avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques. J Histochem Cytochem 29:557–560
Korsrud FR, Brandtzaeg P (1982) Characterization of epithelial elements in human major salivary glands by functional markers. J Histochem Cytochem 30:657–666
Kraehenbuhl JP, Racine L, Jamieson JD (1977) Immunocytochemical localization of secretory protein in bovine pancreatic exocrine cells. J Cell Biol 72:406–423
Kraus FW, Mestecky J (1971) Immunohistochemical localization of amylase, lysozyme and immunoglobulins in the human parotid gland. Arch Oral Biol 16:781–789
Mollenhauer HH (1974) Poststaining sections for electron microscopy. Stain Technol 49:305–308
Nieuw Amerongen AV, Oderkerk CH, Bos Vreugdenhil AP, Roukema PA (1982) Secretory granules of murine salivary glands. A morphological, biochemical and immunochemical study of submandibular and parotid granules. J Biol Buccale 10:11–30
Palade G (1975) Intracellular aspects of the process of protein synthesis. Science 189:347–358
Robinovitch MR, Sreebuy LM, Smuckler EA (1966) Preservation of the secretory granules of rat parotid gland of electron microscopy. Exp Cell Res 42:634–639
Roth J (1982) The protein A-gold (pAg) technique - a qualitative and quantitative approach for antigen localization on thin sections. In: Bullock GR, Petrusz P (eds) Techniques in immunocytochemistry. Academic Press, London, New York, pp 107–133
Roth J, Bendayan M, Orci L (1978) Ultrastructural localization of intracellular antigens by the use of protein A-gold complex. J Histochem Cytochem 26:1074–1081
Rothman SS (1975) Protein transport by the pancreas. Science 190:747–753
Seifert G (1962) Elektronenmikroskopische Befunde an den Speicheldrüsenacini nach Einwirkung von Noradrenalin. Beitr Path Anat 127:111–136
Simson JAV, Spicer SS, Hall BJ (1974) Morphology and cytochemistry of rat salivary gland acinar secretory granules and their alteration by isoproterenol. J Ultrastruct Res 48:465–482
Simson JAV, Dom RM, Sannes PL, Spicer SS (1978) Morphology and cytochemistry of acinar secretory granules in normal and isoproterenol-treated rat submandibular glands. J Microsc 113:185–203
Smith RJ, Frommer J (1972) The starch substrate film method for the localization of amylase activity. J Histochem Cytochem 21:189–190
Tanaka T, Gresik EW, Michelakis AM, Barka T (1980) Immunocytochemical localization of renin in kidneys and submandibular glands of SWR/J and C57BL/6J mice. J Histochem Cytochem 28:1113–1118
Tanaka T, Gresik EW, Barka T (1981) Immunocytochemical localization of amylase in the parotid gland of developing and adult rats. J Histochem Cytochem 29:1189–1195
Wood GS, Warnke R (1981) Suppression of endogenous avidin-binding activity in tissues and its relevance to biotin-avidin detection systems. J Histochem Cytochem 29:1196–1204
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Supported by the Deutsche Forschungsgemeinschaft and by the Hamburger Stiftung zur Förderung der Krebsbekämpfung
Dedicated to Prof Dr. med. Dres. h.c. Wilhelm Doerr, Heidelberg, in honour of his 70th birthday
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Zimmer, K.P., Caselitz, J., Seifert, G. et al. Immunoelectron microscopy of amylase in the human parotid gland. Vichows Archiv A Pathol Anat 404, 187–196 (1984). https://doi.org/10.1007/BF00704063
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DOI: https://doi.org/10.1007/BF00704063