The Histochemical Journal

, Volume 20, Issue 5, pp 276–282 | Cite as

Lectin binding to rat spermatogenic cells: Effects of different fixation methods and proteolytic enzyme treatment

  • R. Malmi
  • K. -O. Söderström
Papers
Received:
Revised:

Summary

The binding of a panel of eight different fluorescein-conjugated lectins to rat spermatogenic cells was investigated. Particular attention was paid to the effects of different fixation methods and proteolytic enzyme digestion on the staining pattern.

Concanavalin A (Con A), wheatgerm agglutinin (WGA), succinylated WGA (s-WGA) and agglutinin from gorse (UEA I) stained the cytoplasm of most germ cells as well as the spermatid acrosome. In contrast, peanut agglutinin (PNA), castor bean agglutinin (RCAI) and soy bean agglutinin (SBA) mainly stained the acrosome. The staining pattern varied depending on the fixation method used. PNA was particularly sensitive to formalin fixation, while SBA, DBA and UEA I showed decreased binding and Con A, WGA, s-WGA and RCA I were insensitive to this type of fixation. Pepsin treatment of the sections before lectin staining caused marked changes in the staining pattern; staining with PNA in formalin-fixed tissue sections was particularly improved but there was also enhanced staining with SBA and horse gram agglutinin (DBA). On the other hand, in Bouin- and particularly in acetone-fixed tissue sections, pepsin treatment decreased the staining with several of the lectins, for example WGA and UEA I.

Keywords

Germ Cell Fixation Method Proteolytic Enzyme Staining Pattern Enzyme Treatment 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in to check access.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. ALLISON, R. T. (1987) The effects of various fixatives on subsequent lectin binding to tissue sections.Histochem. J. 19, 65–74.Google Scholar
  2. BRASITUS, T. A., GOLDFARB, J. P., HSU, K. C. & TANNENBAUM, M. (1982) Characterization of rat colonic cell surface glycoconjugates by fluorescinated lectins: I. Importance of fixation techniques.Histochemistry 76, 197–209.Google Scholar
  3. CURRAN, R. C. & GREGORY, J. (1980) Effects of fixation and processing on immunohistochemical demonstration of immunoglobulin in paraffin sections of tonsil and bone marrow.J. clin. Pathol. 33, 1047–57.Google Scholar
  4. DEBRAY, H., DECOUT, D., STRECKLER, G., SPIK, G. & MONTREUL, J. (1981) Specificity of 12 lectins towards oligosaccharides and glycopeptides related toN-glycosylproteins.Eur. J. Biochem. 117, 41–55.Google Scholar
  5. GOLDSTEIN, I. J. & HAYES, C. E. (1978) The lectins: Carbohydrate binding proteins of plants and animals.Adv. Carbohydr. Chem. Biochem. 35, 127–340.Google Scholar
  6. HOLTHÖFER, H., MIETTINEN, A., PASSIVUO, R., LEHTO, V.-P., LINDER, E., ALFTHAN, O. & VIRTANEN, I. (1983) Cellular origin and differentiation of renal carcinomas: A fluorescence microscopic study with kidney specific antibodies, antiintermediate filament antibodies and lectins.Lab. Invest. 49, 317–26.Google Scholar
  7. HSU, S. M. & REE, H. J. (1983) Histochemical studies on lectin binding in reactive lymphoid tissue.J. Histochem. Cytochem. 31, 538–46.Google Scholar
  8. JEFFREY, I. J. M., MOSLEY, S. M., JONES, C. J. P. & STODDART, R. W. (1987) Proteolysis and lectin histochemistry.Histochem. J. 19, 269–75.Google Scholar
  9. LEATHEM, A. (1986) Lectin histochemistry. InImmunocytochemistry. Modern Methods and Applications (edited by POLAK, J. M. & VAN NOORDEN, S.), pp. 167–87. Bristol: J. Wright & Sons.Google Scholar
  10. MALMI, R., KALLAJOKI, M. & SUOMINEN, J. (1987) Distribution of glycoconjugates in human testis. A histochemical study using fluorescein and rhodamine conjugated lectins.Andrologia 19, 322–32.Google Scholar
  11. MONSIGNY, M., SENE, C., OBRENOVITCH, A., ROCHE, A. C., DELMOTTE, F. & BOSHETTI, E. (1979) Properties of succinylated wheat germ agglutinin.Eur. J. Biochem. 98, 39–45.Google Scholar
  12. PEARSE, A. G. E. (1960)Histochemistry: Theoretical and Applied, pp. 53–74. London: Churchill.Google Scholar
  13. PUCHTLER, H. & MELOAN, S. N. (1985) On the chemistry of formaldehyde fixation and its effects on immunohistochemical reactions.Histochemistry 82, 201–4.Google Scholar
  14. ROMEIS, B. (1948)Mikroskopische Technik, 15 Aufl. Leibniz.Google Scholar
  15. RITTMAN, I. C. & MACKENZIE, B. R. (1983) Effect of histological processing on lectin binding patterns in oral mucosa and skin.Histochem. J. 15, 467–74.Google Scholar
  16. SÖDERSTRÖM, K.-O., MALMI, R. & KARJALAINEN, K. (1984) Binding of fluorescein isothiocyanate conjugated lectins to rat spermatogenic cells in tissue sections: Enhancement of lectin fluorescence obtained by fixation in Bouin's fluid.Histochemstry 80, 475–9.Google Scholar
  17. SÖDERSTRÖM, K.-O. (1987) Lectin binding to formalin fixed paraffin sections of human endometrium.Int. J. Gyn. Path. 6, 55–65.Google Scholar
  18. WALKER, J. F. (1964)Formaldehyde, 3rd edn. New York: Reinhold.Google Scholar

Copyright information

© Chapman and Hall Ltd 1988

Authors and Affiliations

  • R. Malmi
    • 1
  • K. -O. Söderström
    • 1
  1. 1.Department of PathologyUniversity of TurkuTurkuFinland

Personalised recommendations