Skip to main content
Log in

The use of trypsin to improve the localization of immunoglobulins in semi-thin frozen sections of the mammary gland

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

Summary

Frozen sections, 0.5 μm thick, of the lactating mouse mammary gland have been used to localize immunoglobulins A and G and serum albumin throughout the connective tissue stroma, in the lumina of blood vessels, in milk stored in the alveoli and in the lateral spaces between adjacent epithelial cells. In addition, the immunoglobulins were localized to their specific plasma cells in the connective tissue stroma. Serum albumin was further identified within the mammary epithelial cells as small spots of fluorescence scattered throughout the cytoplasm. The immunoglobulins were not localized within these cells in untreated sections, but in sections treated with trypsin and Soybean trypsin inhibitor, it was possible to identify a similar distribution to that for serum albumin. The spots of fluorescence representing the intracellular localization of the immunoglobulins and serum albumin were frequently found in association with the periphery of intracellular lipid droplets.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Brown, P. J., Bourne, F. J. &Denny, H. R. (1975) Immunoglobulin-containing cells in pig mammary gland.J. Anat. 120, 329–35.

    Google Scholar 

  • Dixon, F. J., Weigle, W. O. &Vazquez, J. J. (1961) Metabolism and mammary secretion of serum proteins in the cow.Lab. Invest. 10, 216–37.

    Google Scholar 

  • Finley, J. C. W. &Petrusz, P. (1982) The use of proteolytic enzymes for improved localisation of tissue antigens with immunocytochemistry. InTechniques in Immunocytochemistry (edited byBullock, G. R. andPetrusz, P.), Vol. I, pp. 239–49. London: Academic Press.

    Google Scholar 

  • Fujimura, M., Chen, S. T., Sudo, T., Kyuama, T. &Saito, T. (1981) Cellular sites of immunoglobulins. VII. Localisation of immunoglobulins in female mammary gland.Acta histochem. cytochem. 14, 163–7.

    Google Scholar 

  • Hogan, D. L. &Smith, G. H. (1982) Unconventional application of standard light and electron immunocytochemical analysis to aldehyde-fixed, araldite-embedded tissues.J. Histochem. Cytochem. 30, 1301–6.

    Google Scholar 

  • Koldovsky, O. (1980) Hormones in milk.Life Sci. 26, 1833–6.

    Google Scholar 

  • Kraehenbuhl, J. P. (1978) Transport of immunoglobulins across epithelia. InTransport of Macromolecules in Cellular Systems (edited bySilverstein, S. C.), pp. 213–28. Berlin: Dahlem Konferensen.

  • Kraehenbuhl, J. P., Racine, L. &Galardy, R. E. (1975) Localisation of secretory IgA, secretory component, and alpha chain in the mammary gland of lactating rabbits by immunoelectron microscopy.Ann. N.Y. Acad. Sci. 254, 190–201.

    Google Scholar 

  • Kuhn, L. C. &Kraehenbuhl, J. P. (1982) The sacrificial receptor-translocation of polymeric IgA across epithelia.Trends Biochem. 7, 299–302.

    Google Scholar 

  • Lee, C. G., Ladds, P. W. &Watson, D. L. (1978) Immunocyte populations in the mammary gland of the rat at different stages of pregnancy and lactation.Res. Vet. Sci. 24, 322–7.

    Google Scholar 

  • Nolin, J. M. &Witorsch, R. J. (1976) Detection of endogenous immunoreactive prolactin in rat mammary epithelial cells during lactation.Endocrinology 99, 949–58.

    Google Scholar 

  • Pitelka, D. R. (1978) Cell contacts in the mammary gland. InLactation: a comprehensive Treatise (edited byLarson, B. L.), Vol. IV, pp. 41–66. New York: Academic Press.

    Google Scholar 

  • Richards, R. C. (1979) Milk secretion: with special reference to its content of hormones, enzymes and immunoreactive components. InOxford Reviews of Reproductive Biology (edited byFinn, C. A.), Vol. I, pp. 262–82. Oxford: Clarendon Press.

    Google Scholar 

  • Richards, R. C. (1981) The immunofluorescent localisation of immunoglobulins and serum albumin in the lactating mammary gland.Cell Biol. Int. Rep. Suppl. A. 5, P46.

    Google Scholar 

  • Richards, R. C. (1983) The use of semi-thin frozen sections in immunocytochemistry. InImmunocytochemistry. Practical Applications in Pathology and Biology (edited byPolak, J. M. andVan Noorden, S.), pp. 69–81. London: Wright PSG.

    Google Scholar 

  • Sasaki, M., Larson, B. L. &Nelson, D. R. (1977) Kinetic analysis of the binding of immunoglobulin IgG1 and IgG2 to bovine mammary cells.Biochem. Biophys. Acta 497, 160–70.

    Google Scholar 

  • Silverstein, S. C., Steinman, R. M. &Cohn, Z. A. (1977) Endocytosis.Ann. Rev. Biochem. 46, 669–772.

    Google Scholar 

  • Syre, G. &Sehn, M. (1981) Intracellular storage of IgA and secretory component in carcinomas of the female breast.Virchows Arch. Path. Anat. Physiol. 393, 315–20.

    Google Scholar 

  • Tokuyasu, K. T. (1980) Immunocytochemistry on ultrathin frozen sections.Histochem. J. 12, 381–403.

    Google Scholar 

  • Weisz-Carrington, P., Roux, M. E. &Lamm, M. E. (1977) Plasma cells and epithelial immunoglobulins in the mouse mammary gland during pregnancy and lactation.J. Immun. 119, 1306–9.

    Google Scholar 

  • Weisz-Carrington, P., Roux, M. E., McWilliams, M., Phillips-Quagliata, J. M. &Lamm, M. E. (1978) Hormonal induction of the secretory immune system in the mammary gland.Proc. Nat. Acad. Sci. USA 75, 2928–32.

    Google Scholar 

  • Wooding, F. B. P. (1971) The mechanism of secretion of the milk fat globule.J. Cell Sci. 9, 805–21.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Richards, R.C. The use of trypsin to improve the localization of immunoglobulins in semi-thin frozen sections of the mammary gland. Histochem J 16, 565–572 (1984). https://doi.org/10.1007/BF01041356

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01041356

Keywords

Navigation