The Histochemical Journal

, Volume 10, Issue 4, pp 425–433 | Cite as

Endogenous peroxidase in mast cells localized with a semipermeable membrane technique

  • K. N. Christie
  • P. J. Stoward
Papers

Synopsis

Hamster mast cells have been found to give strong peroxidatic reactions at pH 5, 7.5 and 10 when sections of skeletal muscle are incubated for 2.5 h in the dark at room temperature on semipermeable membranes covering a gelled incubation medium consisting of 0.01% hydrogen peroxide, 5.5 mM diaminobenzidine and 1.36% agar dissolved in Universal buffer. The techniques is very efficient: with it, all mast cells react in marked contrast to the negative reaction they usually give with conventional techniques.

The peroxidatic reactions are abolished if tissues are perfused beforehand with either aminotriazole or KCN but not if these inhibitors are incorporated in the gelled incubation medium. This and other evidence suggests that the mast cell reactions are not due to either catalase or haemoglobin adsorbed onto mast cell granules from lysed red blood cells.

Skeletal muscle fibres do not exhibit any visible-peroxidase activity with the membrane technique.

Keywords

Skeletal Muscle Mast Cell Catalase Cell Granule Diaminobenzidine 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Christie, K. N. &Stoward, P. J. (1977a) Endogenous peroxidase in muscle fibres.Proc. Roy. micr. Soc. 12, 120–1.Google Scholar
  2. Christie, K. N. & Stoward, P. J. (1977b). Cytochemical localisation of endogenous peroxidase in the sarcoplasmic reticulum of hamster skeletal muscle.Riv. Istochim. Norm. Patol. (in press).Google Scholar
  3. Desaga, J. F. (1972). Das Nachweiss von Myeloperoxydase in jungen Gewebsmaztzellen der Ratte.Klin. Wschr. 50, 444–5.Google Scholar
  4. Dvorak, A. M., Dvorak, H. F. &Karnovsky, M. J. (1972). Cytochemical localisation of peroxidase activity in the developing erythrocyte.Am. J. Path. 67, 303–15.Google Scholar
  5. Fahimi, H. D. (1970). The fine structural localisation of endogenous and exogenous peroxidase activity in Kupffer cells of rat liver.J. Cell Biol. 47, 247–62.Google Scholar
  6. Fahimi, H. D. (1973). Diffusion artifacts in cytochemistry of catalase.J. Histochem. Cytochem. 21, 999–1009.Google Scholar
  7. Hand, A. R. (1974). Peroxisomes (microbodies) in striated muscle cells.J. Histochem. Cytochem. 22, 207–9.Google Scholar
  8. Herzog, V. &Fahimi, H. D. (1973). A new colorimetric assay for peroxidase using 3,3′-diaminobenzidine as hydrogen donor.Analyt. Biochem. 55, 554–62.Google Scholar
  9. Herzog, V. &Fahimi, H. D. (1976). Intracellular distinction between peroxidase and catalase in exocrine cells of rat lacrimal gland: a biochemical and cytochemical study.Histochemistry 46, 273–86.Google Scholar
  10. Lojda, Z., Gossrau, R. &Schiebler, T. H. (1976).Enzym-histochemische Methoden. Berlin, Heidelberg & New York: Springer-Verlag.Google Scholar
  11. Meijer, A. E. F. H. (1972). Semipermeable membranes for improving the histochemical demonstration of enzyme activities in tissue sections. 1. Acid phosphatase.Histochemie 30, 31–9.Google Scholar
  12. Meijer, A. E. F. H. & deVries, G. P. (1974). Semipermeable membranes for improving the histochemical demonstration of enzyme activities in tissue sections. IV. Glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase (decarboxylating).Histochemistry 40, 349–59.Google Scholar
  13. Novikoff, A. B., Beard, M. E., Albala, A., Sheid, B., Quintana, N. &Biempica, L. (1971). Localisation of endogenous peroxidases in animal tissues.J. Microscopie 12, 381–404.Google Scholar
  14. Okun, M. R., Hamada, G., Donnellan, B. &Lever, W. F. (1971). Latency of the Dopa reaction in mast cells.Histochemie 25, 1–8.Google Scholar
  15. Okun, M. R., Edelstein, L. M., Patel, R. P. &Donnellan, B. (1973). A revised concept of mammalian melanogenesis: the possible synergistic functions of aerobic dopa oxidase and peroxidase. A review.Yale J. Biol. Med. 46, 535–40.Google Scholar
  16. Rechcigl, M. &Evans, W. H. (1963). Role of catalase and peroxidase in the metabolism of leucocytes.Nature 199, 1001–2.Google Scholar
  17. Robbins, D., Fahimi, H. D. &Cotran, R. S. (1971). Fine structural cytochemical localisation of peroxidase activity in rat peritoneal cells: mononuclear cells, eosinophils and mast cells.J. Histochem. Cytochem. 19, 571–5.Google Scholar
  18. Selye, H. (1965).The Mast Cells, p. 50. Washington: Butterworths.Google Scholar
  19. Somer, J. R. &Waugh, R. A. (1976). The ultrastructure of the mammalian cardiac inuscle cell—with special emphasis on the tubular membrane systems.Am. J. Path. 82, 191–221.Google Scholar

Copyright information

© Chapman and Hall Ltd 1978

Authors and Affiliations

  • K. N. Christie
    • 1
  • P. J. Stoward
    • 1
  1. 1.Department of AnatomyUniversity of DundeeScotland

Personalised recommendations