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. 1972 Feb;66(2):277–294.

Ultrastructural Localization of Peroxidase Activity in Human Platelets and Megakaryocytes

Janine Breton-Gorius, Josette Guichard
PMCID: PMC2032483  PMID: 5009974

Abstract

Normal human platelets and megakaryocytes were examined for peroxidase activity by the diaminobenzidine (DAB) cytochemical technic. When the fixation and the incubation were adequate, a strong reaction was present in the dense tubular system of platelets suspended in plasma or spread on carbon. The black reaction product was ascribed to enzyme activity, since the reaction was completely eliminated when H2O2 or DAB were omitted, or when H2O2 was in excess. In addition, the reaction was inhibited by aminotriazole, cyanide and azide. In the human megakaryocytes, the reaction was localized in the endoplasmic reticulum including the perinuclear envelope. The Golgi complex and the clear vacuolar system were negative for the reaction. After platelet release, the reaction was always seen in the perinuclear space. The nature and function of the enzyme, as well as its possible relationships with catalase, are discussed.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bainton D. F., Farquhar M. G. Differences in enzyme content of azurophil and specific granules of polymorphonuclear leukocytes. II. Cytochemistry and electron microscopy of bone marrow cells. J Cell Biol. 1968 Nov;39(2):299–317. doi: 10.1083/jcb.39.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bainton D. F., Farquhar M. G. Segregation and packaging of granule enzymes in eosinophilic leukocytes. J Cell Biol. 1970 Apr;45(1):54–73. doi: 10.1083/jcb.45.1.54. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beard M. E., Novikoff A. B. Distribution of peroxisomes (microbodies) in the nephron of the rat: a cytochemical study. J Cell Biol. 1969 Aug;42(2):501–518. doi: 10.1083/jcb.42.2.501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Behnke O. An electron microscope study of the megacaryocyte of the rat bone marrow. I. The development of the demarcation membrane system and the platelet surface coat. J Ultrastruct Res. 1968 Sep;24(5):412–433. doi: 10.1016/s0022-5320(68)80046-2. [DOI] [PubMed] [Google Scholar]
  5. Behnke O. An electron microscope study of the rat megacaryocyte. II. Some aspects of platelet release and microtubules. J Ultrastruct Res. 1969 Jan;26(1):111–129. doi: 10.1016/s0022-5320(69)90039-2. [DOI] [PubMed] [Google Scholar]
  6. Behnke O. Electron microscopic observations on the membrane systems of the rat blood platelet. Anat Rec. 1967 Jun;158(2):121–137. doi: 10.1002/ar.1091580203. [DOI] [PubMed] [Google Scholar]
  7. Behnke O. Nonspecific deposition of lead in experiments on fine structural localization of enzymatic activity of rat blood platelets. J Histochem Cytochem. 1966 May;14(5):432–433. doi: 10.1177/14.5.432. [DOI] [PubMed] [Google Scholar]
  8. Bessis M., Breton-Gorius J. Les microtubules et les fibrilles dans les plaquettes étalées. Nouv Rev Fr Hematol. 1965 Jul-Aug;5(4):657–662. [PubMed] [Google Scholar]
  9. Bloom G. D., Carlsö B., Kumlien A. Cytochemical localization of peroxidase acitivity in the submandibular salivary gland of the hamster. Histochemie. 1970;22(4):294–301. doi: 10.1007/BF00277457. [DOI] [PubMed] [Google Scholar]
  10. Breton-Gorius J., Guichard J. Etude au microscope électronique de la localisation des peroxydases dans les cellules DE LA MOELLE OSSEUSE HUMAINE. Nouv Rev Fr Hematol. 1969 Sep-Oct;9(5):678–687. [PubMed] [Google Scholar]
  11. Breton-Gorius J. Utilisation de la diaminobenzidine pour la mise en évidence, au microscope électronique, de l'hémoglobine intracellulaire. La réactivité des différents organelles des érythroblastes. Nouv Rev Fr Hematol. 1970 Mar-Apr;10(2):243–256. [PubMed] [Google Scholar]
  12. Brökelmann J., Fawcett D. W. The localization of endogenous peroxidase in the rat uterus and its induction by estradiol. Biol Reprod. 1969 Apr;1(1):59–71. doi: 10.1095/biolreprod1.1.59. [DOI] [PubMed] [Google Scholar]
  13. Cotran R. S., Litt M. Ultrastructural localization of horseradish peroxidase and endogenous peroxidase activity in guinea pig peritoneal macrophages. J Immunol. 1970 Dec;105(6):1536–1546. [PubMed] [Google Scholar]
  14. Da Prada M., Pletscher A., Tranzer J. P., Knuchel H. Subcellular localization of 5-hydroxytryptamine and histamine in blood platelets. Nature. 1967 Dec 30;216(5122):1315–1317. doi: 10.1038/2161315a0. [DOI] [PubMed] [Google Scholar]
  15. Dunn W. B., Hardin J. H., Spicer S. S. Ultrastructural localization of myeloperoxidase in human neutrophil and rabbit heterophil and eosinophil leukocytes. Blood. 1968 Dec;32(6):935–944. [PubMed] [Google Scholar]
  16. Fahimi H. D. Cytochemical localization of peroxidase activity in rat hepatic microbodies (peroxisomes). J Histochem Cytochem. 1968 Aug;16(8):547–550. doi: 10.1177/16.8.547. [DOI] [PubMed] [Google Scholar]
  17. Fahimi H. D. Cytochemical localization of peroxidatic activity of catalase in rat hepatic microbodies (peroxisomes). J Cell Biol. 1969 Nov;43(2):275–288. doi: 10.1083/jcb.43.2.275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Fahimi H. D. The fine structural localization of endogenous and exogenous peroxidase activity in Kupffer cells of rat liver. J Cell Biol. 1970 Oct;47(1):247–262. doi: 10.1083/jcb.47.1.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Frederick S. E., Newcomb E. H. Cytochemical localization of catalase in leaf microbodies (peroxisomes). J Cell Biol. 1969 Nov;43(2):343–353. doi: 10.1083/jcb.43.2.343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Graham R. C., Jr, Karnovsky M. J. The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem. 1966 Apr;14(4):291–302. doi: 10.1177/14.4.291. [DOI] [PubMed] [Google Scholar]
  21. Herzog V., Miller F. Die Lokalisation endogener Peroxydase in der Glandula parotis der Ratte. Z Zellforsch Mikrosk Anat. 1970;107(3):403–420. [PubMed] [Google Scholar]
  22. MORITA H., ASADA T. Etude morphologique des plaquettes sanguines de l'homme. Sang. 1957;28(9):827–854. [PubMed] [Google Scholar]
  23. Marcus A. J., Zucker-Franklin D., Safier L. B., Ullman H. L. Studies on human platelet granules and membranes. J Clin Invest. 1966 Jan;45(1):14–28. doi: 10.1172/JCI105318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Maupin B., Saint-Blancard J. Introduction à l'étude des enzymes plaquettaires. Nouv Rev Fr Hematol. 1966 Mar-Apr;6(2):255–264. [PubMed] [Google Scholar]
  25. Novikoff A. B., Goldfischer S. Visualization of peroxisomes (microbodies) and mitochondria with diaminobenzidine. J Histochem Cytochem. 1969 Oct;17(10):675–680. doi: 10.1177/17.10.675. [DOI] [PubMed] [Google Scholar]
  26. Roels F., Schiller B., Goldfischer S. Microbodies (peroxisomes) in the toad, Bufo marinus. A cytochemical study. Z Zellforsch Mikrosk Anat. 1970;108(2):135–149. doi: 10.1007/BF00335292. [DOI] [PubMed] [Google Scholar]
  27. SABATINI D. D., BENSCH K., BARRNETT R. J. Cytochemistry and electron microscopy. The preservation of cellular ultrastructure and enzymatic activity by aldehyde fixation. J Cell Biol. 1963 Apr;17:19–58. doi: 10.1083/jcb.17.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Seligman A. M., Karnovsky M. J., Wasserkrug H. L., Hanker J. S. Nondroplet ultrastructural demonstration of cytochrome oxidase activity with a polymerizing osmiophilic reagent, diaminobenzidine (DAB). J Cell Biol. 1968 Jul;38(1):1–14. doi: 10.1083/jcb.38.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Spicer S. S., Greene W. B., Hardin J. H. Ultrastructural localization of acid mucosubstance and antimonate-precipitable cation in human and rabbit platelets and megakaryocytes. J Histochem Cytochem. 1969 Dec;17(12):781–792. doi: 10.1177/17.12.781. [DOI] [PubMed] [Google Scholar]
  30. Strum J. M., Karnovsky M. J. Aminotriazole goiter. Fine structure and localization of thyroid peroxidase activity. Lab Invest. 1971 Jan;24(1):1–12. [PubMed] [Google Scholar]
  31. Strum J. M., Karnovsky M. J. Ultrastructural localization of peroxidase in submaxillary acinar cells. J Ultrastruct Res. 1970 May;31(3):323–336. doi: 10.1016/s0022-5320(70)90135-8. [DOI] [PubMed] [Google Scholar]
  32. Venkatachalam M. A., Soltani M. H., Fahimi H. D. Fine structural localization of peroxidase activity in the epithelium of large intestine of rat. J Cell Biol. 1970 Jul;46(1):168–173. doi: 10.1083/jcb.46.1.168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Vigil E. L. Intracellular localization of catalase (peroxidatic) activity in plant microbodies. J Histochem Cytochem. 1969 Jun;17(6):425–428. doi: 10.1177/17.6.425. [DOI] [PubMed] [Google Scholar]
  34. White J. G. Effects of colchicine and vinca alkaloids on human platelets. II. Changes in the dense tubular system and formation of an unusual inclusion in incubated cells. Am J Pathol. 1968 Sep;53(3):447–461. [PMC free article] [PubMed] [Google Scholar]
  35. White J. G. The submembrane filaments of blood platelets. Am J Pathol. 1969 Aug;56(2):267–277. [PMC free article] [PubMed] [Google Scholar]
  36. Wood R. L., Legg P. G. Peroxidase activity in rat liver microbodies after amino-triazole inhibition. J Cell Biol. 1970 Jun;45(3):576–585. doi: 10.1083/jcb.45.3.576. [DOI] [PMC free article] [PubMed] [Google Scholar]

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