Skip to main content
SpringerLink
Log in

Concentration of amylase along its secretory pathway in the pancreatic acinar cell as revealed by high resolution immunocytochemistry

  • Papers
  • Published:
The Histochemical Journal Aims and scope Submit manuscript

Summary

The modified protein A-gold immunocytochemical technique was applied to the localization of amylase in rat pancreatic acinar cells. Due to the good ultrastructural preservation of the cellular organelles obtained on glutaraldehyde-fixed, osmium tetroxide-postfixed tissue, the labelling was detected with high resolution over the cisternae of the rough endoplasmic reticulum (RER), the Golgi apparatus, the condensing vacuoles, the immature ‘pre-zymogen’ granules, and the mature zymogen granules. Over the Golgi area, the labelling was present over the transitional elements of the endoplasmic reticulum, some of the smooth vesicular structures at thecis- andtrans-faces and all the different Golgi cisternae. The acid phosphatase-positive rigidtrans-cisternae as well as the coated vesicles were either negative or weakly labelled. Quantitative evaluations of the degree of labelling demonstrated an increasing intensity which progresses from the RER, through the Golgi, to the zymogen granules and have identified the sites where protein concentration occurs. The results obtained have thus demonstrated that amylase is processed through the conventional RER-Golgi-granule secretory pathway in the pancreatic acinar cells. In addition a concomitance has been found between some sites where protein concentration occurs: thetrans-most Golgi cisternae, the condensing vacuoles, the pre- and the mature zymogen granules, and the presence of actin at the level of the limiting membranes of these same organelles as reported previously (Bendayan, 1983). This suggests that beside their possible role in transport and release of secretory products, contractile proteins may also be involved in the process of protein concentration.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Bendayan, M. (1981) Ultrastructural localization of nucleic acids by the use of enzyme-gold complexes.J. Histochem. Cytochem. 29, 531–41.

    Google Scholar 

  • Bendayan, M. (1982a) Double immunocytochemical labeling applying the protein A-gold technique.J. Histochem. Cytochem. 30, 81–5.

    Google Scholar 

  • Bendayan, M. (1982b) Ultrastructural localization of nucleic acids by the use of enzyme-gold complexes: Influence of fixation and embedding.Biol. Cell 43, 153–6.

    Google Scholar 

  • Bendayan, M. (1983) Ultrastructural localization of actin in muscle, epithelial and secretory cells by applying the protein A-gold immunocytochemical technique.Histochem. J. 15, 39–58.

    Google Scholar 

  • Bendayan, M. &Ito, S. (1979) Immunohistochemical localization of exocrine enzymes in normal rat pancreas.J. Histochem. Cytochem. 27, 1029–34.

    Google Scholar 

  • Bendayan, M., Marceau, N., Beaudoin, A. R. &Trifaro, J. M. (1982) Immunocytochemical localization of actin in the pancreatic exocrine cell.J. Histochem. Cytochem. 30, 1075–8.

    Google Scholar 

  • Bendayan, M. &Ørstavik, T. B. (1982) Immunocytochemical localization of kallikrein in the rat exocrine pancreas.J. Histochem. Cytochem. 30, 58–66.

    Google Scholar 

  • Bendayan, M., Roth, J., Perrelet, A. &Orci, L. (1980) Quantitative immunocytochemical localization of pancreatic secretory proteins in sub-cellular compartments of the rat acinar cell.J. Histochem. Cytochem. 28, 149–60.

    Google Scholar 

  • Bendayan, M. &Shore, G. C. (1982) Immunocytochemical localization of mitochondrial proteins in the rat hepatocyte.J. Histochem. Cytochem. 30, 139–47.

    Google Scholar 

  • Bendayan, M. &Zollinger, M. (1983) Ultrastructural localization of antigenic sites on osmium-fixed tissues applying the protein A-gold technique.J. Histochem. Cytochem. 31, 101–9.

    Google Scholar 

  • Broadwell, R. C., Oliver, C. &Brightman, M. W. (1979) Localization of neurophysin within organelles associated with protein synthesis and packaging in the hypothalamo-neurohypophysal system: an immunocytochemical study.Proc. natn. Acad. Sci. U. S. A. 76, 5999–6003.

    Google Scholar 

  • Caro, L. G. &Palade, G. E. (1964) Protein synthesis, storage and discharge in the pancreatic exocrine cell. An autoradiographic study.J. Cell Biol. 20, 473–95.

    Google Scholar 

  • Clemente, F. &Meldolesi, J. (1975) Calcium in pancreatic secretion. I. Subcellular distribution of calcium and magnesium in the exocrine pancreas of the guinea pig.J. Cell Biol. 65, 88–102.

    Google Scholar 

  • Deduve, C. (1969) Lysosomes in biology and pathology. InThe Lysosome in Retrospect (edited byDingle, J. T. andFell, H. B.), Vol. 1, pp. 3–40. Amsterdam: North-Holland Publishing Co.

    Google Scholar 

  • Farquhar, M. G. &Palade, G. E. (1981) The Golgi apparatus (complex)-(1954–1981)-from artifact to center stage.J. Cell Biol. 91, 77s-103s.

    Google Scholar 

  • Forsgreen, A. &Sjöquist, J. (1966) “Protein A” fromS. aureus. I. Pseudo-immune reaction with human γ-globulin.J. Immun. 97, 822–7.

    Google Scholar 

  • Frens, G. (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold solutions.Nature Phys. Sci. 241, 20–2.

    Google Scholar 

  • Geuze, J. J., Slot, J. W. &Tokuyasu, K. T. (1979) Immunocytochemical localization of amylase and chymotrypsinogen in the exocrine pancreatic cell with special attention to the Golgi complex.J. Cell Biol. 82, 697–707.

    Google Scholar 

  • Goudswaard, J., Van Der Donk, J. A., Noordzij, A., Van Dam, R. H. &Vaerman, J. P. (1978) Protein A reactivity of various mammalian immunoglobulins.Scand. J. Immun. 8, 21–8.

    Google Scholar 

  • Greene, L. J., Hirs, C. H. &Palade, G. E. (1963) On the protein composition of bovine pancreatic zymogen granules.J. biol. Chem. 238, 2054–70.

    Google Scholar 

  • Hand, A. R. &Oliver, C. (1977) Cytochemical studies of GERL and its role in secretory granule formation in exocrine cells.Histochem. J. 9, 375–92.

    Google Scholar 

  • Hand, A. R. &Oliver, C. (1981) The Golgiapparatus: protein transport and packaging in secretory cells InMethods in Cell Biology (edited byHand, A. R. andOliver, C.), Vol. 23, pp. 137–53. New York: Academic Press.

    Google Scholar 

  • Hansen, L. J., Mangkornkanok-Mark, M. &Reddy, J. K. (1981) Immunohistochemical localization of pancreatic exocrine enzymes in normal and neoplastic pancreatic acinar epithelium of rat.J. Histochem. Cytochem. 29, 309–13.

    Google Scholar 

  • Jamieson, J. D. (1981) Summary and perspectives. InMethods in Cell Biology (edited byHand, A. R. andOliver, C.), Vol. 23, pp. 547–55. New York: Academic Press.

    Google Scholar 

  • Jamieson, J. D. &Palade, G. E. (1967) Intracellular transport of secetory proteins in the pancreatic exocrine cell. II. Transport to condensing vacuoles and zymogen granules.J. Cell Biol. 34, 597–615.

    Google Scholar 

  • Jamieson, J. D. &Palade, G. E. (1971a) Condensing vacuole conversion and zymogen granule discharge in pancreatic exocrine cells: metabolic studies.J. Cell Biol. 48, 503–22.

    Google Scholar 

  • Jamieson, J. D. &Palade, G. E. (1971b) Synthesis, intracellular transport and discharge of secretory proteins in stimulated pancreatic exocrine cells.J. Cell Biol. 50, 135–58.

    Google Scholar 

  • Kraehenbuhl, J. P., Racine, L. &Jamieson, J. D. (1977) Immunocytochemical localization of secretory proteins in bovine pancreatic exocrine cells.J. Cell Biol. 72, 406–23.

    Google Scholar 

  • Marchis-Mouren, F. (1965) Etude comparée de l'équipement enzymatique du suc pancéatique de diverses espèces.Bull Soc. Chim. Biol. 47, 2207–17.

    Google Scholar 

  • Novikoff, A. B., Mori, M., Quintana, N. &Yam, A. (1977) Studies of the secretory process in the mammalian exocrine pancreas. I. The condensing vacuoles.J. Cell Biol. 75, 148–65.

    Google Scholar 

  • Novikoff, P. M. &Yam, A. (1978) Sites of lipoprotein particles in normal rat hepatocytes.J. Cell Biol. 76, 1–11.

    Google Scholar 

  • Oliver, C. (1980) Cytochemical localization of acid phosphatase and trimetaphosphatase activities in exocrine acinar cells.J. Histochem. Cytochem. 28, 78–81.

    Google Scholar 

  • Ono, T. (1980) Histochemical and immunohistochemical study of lipase in mouse pancreas.Acta histochem. cytochem. 13, 553–70.

    Google Scholar 

  • Palade, G. E. (1975) Intracellular aspects of the process of protein synthesis.Science N. Y. 189, 347–58.

    Google Scholar 

  • Reggio, H. A. &Palade, G. E. (1978) Sulfated compounds in the zymogen granules of the guinea pig pancreas.J. Cell Biol. 77, 288–314.

    Google Scholar 

  • Roth, J. (1982) New approaches forin situ localization of antigens and glycoconjugates on thin sections: the protein A-gold technique and the lectin-colloidal gold marker system.Proc. int. Congress Elect. Microsc. 3, 245–52.

    Google Scholar 

  • Roth, J., Bendayan, M. &Orci, L. (1978) Ultrastructural localization of intracellular antigens by the use of protein A-gold complex.J. Histochem. Cytochem. 26, 1074–81.

    Google Scholar 

  • Roth, J. &Berger, E. G. (1982) Immunocytochemical localization of galactosyltransferase in Hela cells: codistribution with thiamine pyrophosphatase intrans-Golgi cisternae.J. Cell Biol. 93, 223–9.

    Google Scholar 

  • Rothman, J. E. (1981) The Golgi apparatus: two organelles in tandem.Science N. Y. 213, 1212–9.

    Google Scholar 

  • Scheele, G. A. (1974) Two-dimensional gel analysis of soluble proteins.J. biol. Chem. 250, 5375–85.

    Google Scholar 

  • Scheele, G. A., Palade, G. E. &Tartakoff, A. M. (1978) Cell fractionation studies on the guinea pig pancreas: redistribution of exocrine proteins during tissue homogenization.J. Cell Biol. 78, 110–30.

    Google Scholar 

  • Siekevitz, P. &Palade, G. E. (1960) A cytochemical study on the pancreas of the guinea pig. V.In vivo incorporation of leucine-1-C14 into the chymotrypsinogen of various cell fractions.J. biophys. biochem. Cytol. 7, 619–30.

    Google Scholar 

  • Smith, R. E. &Farquhar, M. G. (1966) Lysosome function in the regulation of the secretory process in cells of the anterior pituitary gland.J. Cell Biol. 31, 319–47.

    Google Scholar 

  • Tang, X. M., Lalli, M. F. &Clermont, Y. (1982) A cytochemical study of the Golgi apparatus of the spermatid during spermiogenesis in the rat.Am. J. Anat. 163, 283–94.

    Google Scholar 

  • Tartakoff, A. M. (1980) The Golgi complex: crossroads for vesicular traffic.Int. Rev. Exp. Path. 22, 227–51.

    Google Scholar 

  • Tartakoff, A., Greene, L. J. &Palade, G. E. (1974) Studies on the guinea pig pancreas. Fractionation and partial characterization of exocrine proteins.J. biol. Chem. 249, 7420–31.

    Google Scholar 

  • van Heyningen, H. E. (1964) Secretion of protein by acinar cells of the rat pancreas, as studied by electron microscopic radioautography.Anat. Rec. 148, 485–97.

    Google Scholar 

  • Yasuda, K., Suzuki, T. &Takano, K. (1967) Distribution pattern of the pancreatic amylase around Golgi area, demonstrated by means of immunoelectron microscopic method.Okajimas Fol. Anat. Jap. 43, 87–101.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Anatomy, Université de Montréal, C.P. 6128, H3C 3J7, Montréal, Québec, Canada

    Moïse Bendayan

Authors
  1. Moïse Bendayan
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bendayan, M. Concentration of amylase along its secretory pathway in the pancreatic acinar cell as revealed by high resolution immunocytochemistry. Histochem J 16, 85–108 (1984). https://doi.org/10.1007/BF01003438

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01003438

Keywords

Search

Navigation