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. 1972 Jul 1;54(1):141–156. doi: 10.1083/jcb.54.1.141

CYTOCHEMICAL LOCALIZATION OF ACID PHOSPHATASE ACTIVITY IN GRANULE FRACTIONS FROM RABBIT POLYMORPHONUCLEAR LEUKOCYTES

Marilyn G Farquhar 1, Dorothy F Bainton 1, Marco Baggiolini 1, Christian de Duve 1
PMCID: PMC2108860  PMID: 4114163

Abstract

When rabbit peritoneal exudates (97% polymorphonuclear [PMN] leukocytes, 2% mononuclear cells) were fractionated by zonal sedimentation or isopycnic centrifugation, four fractions (A, B, C, and D) were obtained, as reported earlier. "A" consisted largely of PMN azurophil granules, "B" of PMN specific granules, and "D" of membranous elements. The source of the more heterogeneous "C" fraction (containing acid hydrolases) was uncertain. To gain further information on the nature of this fraction, cytochemical tests for acid phosphatase (AcPase) were carried out on the starting cells and on the fractions. In intact PMN, lead phosphate reaction product was found in Golgi complexes, perinuclear cisternae, and some azurophil granules (immature forms or disrupted mature forms) of a few cells. The specifics and the intact azurophils were not reactive. Reaction product was also found within Golgi cisternae, secondary lysosomes, and some of the azurophil granules of mononuclear cells. Observations on the A and B fractions confirmed those in situ regarding the localization of reaction product in disrupted PMN azurophils, its absence from specifics, and the latency of the enzyme activity in intact azurophils. In the C fraction, AcPase was found in three structures (a) Golgi cisternae, (b) dense bodies, and (c) small pleomorphic granules Comparison with the starting cells indicates that the Golgi complexes are probably derived from both PMN leukocytes and mononuclear cells, whereas the remaining elements resemble (in size, shape, and density) secondary lysosomes and azurophil granules of mononuclear cells. The results indicate that the bulk of the cytochemically detectable AcPase present in the C fraction is derived from mononuclear cells, rather than from PMN leukocytes

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Selected References

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  1. Anderson P. J. Purification and quantitation of glutaraldehyde and its effect on several enzyme activities in skeletal muscle. J Histochem Cytochem. 1967 Aug;15(11):652–661. doi: 10.1177/15.11.652. [DOI] [PubMed] [Google Scholar]
  2. Baggiolini M., De Duve C., Masson P. L., Heremans J. F. Association of lactoferrin with specific granules in rabbit heterophil leukocytes. J Exp Med. 1970 Mar 1;131(3):559–570. doi: 10.1084/jem.131.3.559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baggiolini M., Hirsch J. G., De Duve C. Further biochemical and morphological studies of granule fractions from rabbit heterophil leukocytes. J Cell Biol. 1970 Jun;45(3):586–597. doi: 10.1083/jcb.45.3.586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baggiolini M., Hirsch J. G., De Duve C. Resolution of granules from rabbit heterophil leukocytes into distinct populations by zonal sedimentation. J Cell Biol. 1969 Feb;40(2):529–541. doi: 10.1083/jcb.40.2.529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bainton D. F., Farquhar M. G. Differences in enzyme content of azurophil and specific granules of polymorphonuclear leukocytes. I. Histochemical staining of bone marrow smears. J Cell Biol. 1968 Nov;39(2):286–298. doi: 10.1083/jcb.39.2.286. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Bainton D. F., Farquhar M. G. Origin of granules in polymorphonuclear leukocytes. Two types derived from opposite faces of the Golgi complex in developing granulocytes. J Cell Biol. 1966 Feb;28(2):277–301. doi: 10.1083/jcb.28.2.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bainton D. F., Ullyot J. L., Farquhar M. G. The development of neutrophilic polymorphonuclear leukocytes in human bone marrow. J Exp Med. 1971 Oct 1;134(4):907–934. doi: 10.1084/jem.134.4.907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cohn Z. A., Fedorko M. E., Hirsch J. G. The in vitro differentiation of mononuclear phagocytes. V. The formation of macrophage lysosomes. J Exp Med. 1966 Apr 1;123(4):757–766. doi: 10.1084/jem.123.4.757. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cohn Z. A. The structure and function of monocytes and macrophages. Adv Immunol. 1968;9:163–214. doi: 10.1016/s0065-2776(08)60443-5. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Hardin J. H., Spicer S. S. Ultrastructural localization of dialyzed iron-reactive mucosubstance in rabbit heterophils, basophils, and eosinophils. J Cell Biol. 1971 Feb;48(2):368–386. doi: 10.1083/jcb.48.2.368. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Nichols B. A., Bainton D. F., Farquhar M. G. Differentiation of monocytes. Origin, nature, and fate of their azurophil granules. J Cell Biol. 1971 Aug;50(2):498–515. doi: 10.1083/jcb.50.2.498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Seeman P. M., Palade G. E. Acid phosphatase localization in rabbit eosinophils. J Cell Biol. 1967 Sep;34(3):745–756. doi: 10.1083/jcb.34.3.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Wanson J. C., Tielemans L. Morphological and biochemical characteristics of glycogen particles isolated from rabbit polymorphonuclear leukocytes. J Cell Biol. 1971 Jun;49(3):816–829. doi: 10.1083/jcb.49.3.816. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Zeya H. I., Spitznagel J. K. Characterization of cationic protein-bearing granules of polymorphonuclear leukocytes. Lab Invest. 1971 Mar;24(3):229–236. [PubMed] [Google Scholar]
  17. Zeya H. I., Spitznagel J. K. Isolation of polymorphonuclear leukocyte granules from rabbit bone marrow. Lab Invest. 1971 Mar;24(3):237–245. [PubMed] [Google Scholar]

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