Abstract
For ultrastructural localization of acid mucosubstances in rabbit granulocytes, bone marrow and buffy coat specimens were fixed with formalin, glutaraldehyde, or osmium tetroxide, sectioned at 40 µ, and stained with the Rinehart and Abul-Haj solution of dialyzed iron (DI). Heterophils revealed DI staining on the outer surface of the plasma membrane, in the Golgi complex involved in primary granulogenesis, and in primary granules. The intragranular distribution of DI-stained material varied at different stages in the maturation of primary granules. Immature granules of heterophils fixed by any of the three methods contained a peripheral concentric band of DI-positive material; however, fully mature primary granules possessed a core of DI-reactive material in heterophils fixed with osmium tetroxide, but they contained little or no staining in heterophils fixed with formalin or glutaraldehyde. Secondary granules of rabbit heterophils failed to stain with DI. Tertiary granules, observed only in late heterophils, contained distinct DI-positive particles. Basophil granules exhibited intensely DI-stained material distributed in an orderly pattern throughout the granule. In eosinophils, DI staining was localized in the Golgi complex and in the rims of a few immature cytoplasmic granules.
Full Text
The Full Text of this article is available as a PDF (2.1 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- ACKERMAN G. A. Cytochemical properties of the blood basophilic granulocyte. Ann N Y Acad Sci. 1963 Feb 26;103:376–393. doi: 10.1111/j.1749-6632.1963.tb53710.x. [DOI] [PubMed] [Google Scholar]
- ACKERMAN G. A. HISTOCHEMICAL DIFFERENTIATION DURING NEUTROPHIL DEVELOPMENT AND MATURATION. Ann N Y Acad Sci. 1964 Feb 28;113:537–565. doi: 10.1111/j.1749-6632.1964.tb40690.x. [DOI] [PubMed] [Google Scholar]
- BASSIOUNI M. The estimation of heparin and similar substances in human blood and tissues using a combined biological and colorimetric method with paper electrophoretic studies. J Clin Pathol. 1954 Nov;7(4):330–335. doi: 10.1136/jcp.7.4.330. [DOI] [PMC free article] [PubMed] [Google Scholar]
- BEHRENS M., TAUBERT M. Der Nachweis von Heparin in den basophilen Leukocyten. Klin Wochenschr. 1952 Jan 15;30(3-4):76–78. doi: 10.1007/BF01479711. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- 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]
- 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]
- CLAUSEN J., ANDERSEN V. ACID MUCOPOLYSACCHARIDES IN HUMAN LEUCOCYTES. Clin Chim Acta. 1963 Jul;8:505–512. doi: 10.1016/0009-8981(63)90096-2. [DOI] [PubMed] [Google Scholar]
- COHN Z. A., HIRSCH J. G. The isolation and properties of the specific cytoplasmic granules of rabbit polymorphonuclear leucocytes. J Exp Med. 1960 Dec 1;112:983–1004. doi: 10.1084/jem.112.6.983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cook G. M. Chemistry of membranes. Br Med Bull. 1968 May;24(2):118–123. doi: 10.1093/oxfordjournals.bmb.a070611. [DOI] [PubMed] [Google Scholar]
- DZIEWIATKOWSKI D. D. Rate of excretion of radioactive sulfur and its concentration in some tissues of the rat after intraperitoneal administration of labeled sodium sulfate. J Biol Chem. 1949 Mar;178(1):197–202. [PubMed] [Google Scholar]
- Dunn W. B., Spicer S. S. Histochemical demonstration of sulfated mucosubstances and cationic proteins in human granulocytes and platelets. J Histochem Cytochem. 1969 Oct;17(10):668–674. doi: 10.1177/17.10.668. [DOI] [PubMed] [Google Scholar]
- FEDORKO M. E., MORSE S. I. ISOLATION, CHARACTERIZATION, AND DISTRIBUTION OF ACID MUCOPOLYSACCHARIDES IN RABBIT LEUCOCYTES. J Exp Med. 1965 Jan 1;121:39–48. doi: 10.1084/jem.121.1.39. [DOI] [PMC free article] [PubMed] [Google Scholar]
- FISHER E. R., LILLIE R. D. The effect of methylation on basophilia. J Histochem Cytochem. 1954 Mar;2(2):81–87. doi: 10.1177/2.2.81. [DOI] [PubMed] [Google Scholar]
- Fedorko M. E. Formation of cytoplasmic granules in human eosinophilic myelocytes: an electron microscope autoradiographic study. Blood. 1968 Feb;31(2):188–194. [PubMed] [Google Scholar]
- Fedorko M. E., Hirsch J. G. Cytoplasmic granule formation in myelocytes. An electron microscope radioautographic study on the mechanism of formation of cytoplasmic granules in rabbit heterophilic myelocytes. J Cell Biol. 1966 May;29(2):307–316. doi: 10.1083/jcb.29.2.307. [DOI] [PMC free article] [PubMed] [Google Scholar]
- GASIC G., BERWICK L. HALE STAIN FOR SIALIC ACID-CONTAINING MUCINS. ADAPTATION TO ELECTRON MICROSCOPY. J Cell Biol. 1963 Oct;19:223–228. doi: 10.1083/jcb.19.1.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
- GREULICH R. C. Utilization of sulfate ion by eosinophile myelocytes in the rat. Exp Cell Res. 1956 Aug;11(1):225–228. doi: 10.1016/0014-4827(56)90211-7. [DOI] [PubMed] [Google Scholar]
- HORN R. G., SPICER S. S. SULFATED MUCOPOLYSACCHARIDE AND BASIC PROTEIN IN CERTAIN GRANULES OF RABBIT LEUKOCYTES. Lab Invest. 1964 Jan;13:1–15. [PubMed] [Google Scholar]
- HORN R. G., SPICER S. S., WETZEL B. K. PHAGOCYTOSIS OF BACTERIA BY HETEROPHIL LEUKOCYTES: ACID AND ALKALINE PHOSPHATASE CYTOCHEMISTRY. Am J Pathol. 1964 Aug;45:327–335. [PMC free article] [PubMed] [Google Scholar]
- Hardin J. H., Spicer S. S. An ultrastructural study of human eosinophil granules: maturational stages and pyroantimonate reactive cation. Am J Anat. 1970 Jul;128(3):283–310. doi: 10.1002/aja.1001280303. [DOI] [PubMed] [Google Scholar]
- Hardin J. H., Spicer S. S., Greene W. B. Ultrastructural localization of antimonate deposits in rabbit heterophil and human neutrophil leukocytes. Lab Invest. 1969 Sep;21(3):214–224. [PubMed] [Google Scholar]
- Hudson G. Eosinophil granules and phosphotungstic acid: an electron microscope study of guinea-pig bone marrow. Exp Cell Res. 1966 Feb;41(2):265–273. doi: 10.1016/s0014-4827(66)80134-9. [DOI] [PubMed] [Google Scholar]
- KERBY G. P. The occurrence of acid mucopolysaccharides in human leukocytes and urine. J Clin Invest. 1955 Dec;34(12):1738–1743. doi: 10.1172/JCI103228. [DOI] [PMC free article] [PubMed] [Google Scholar]
- KORN E. D. Inactivation of lipoprotein lipase by heparinase. J Biol Chem. 1957 Jun;226(2):827–832. [PubMed] [Google Scholar]
- LAJTHA L. G., ELLIS F., OLIVER R. Isotope uptake of individual cells: uptake of S35 sulphate by human bone-marrow cells in vitro. Br J Cancer. 1953 Sep;7(3):401–406. doi: 10.1038/bjc.1953.38. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miller F., Herzog V. Die Lokalisation von Peroxydase und saurer Phosphatase in eosinophilen Leukocyten während der Reifung. Elek. Elektronenmikroskopisch-cytochemische Untersuchungen am Knochenmark von Ratte und Kaninchen. Z Zellforsch Mikrosk Anat. 1969;97(1):84–110. [PubMed] [Google Scholar]
- Neutra M., Leblond C. P. Synthesis of the carbohydrate of mucus in the golgi complex as shown by electron microscope radioautography of goblet cells from rats injected with glucose-H3. J Cell Biol. 1966 Jul;30(1):119–136. doi: 10.1083/jcb.30.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
- ODELL T. T., Jr, TAUSCHE F. G., GUDE W. D. Uptake of radioactive sulfate by elements of the blood and the bone marrow of rats. Am J Physiol. 1955 Mar;180(3):491–494. doi: 10.1152/ajplegacy.1955.180.3.491. [DOI] [PubMed] [Google Scholar]
- Olsson I., Gardell S. Isolation and characterization of glycosaminoglycans from human leukocytes and platelets. Biochim Biophys Acta. 1967 Jul 25;141(2):348–357. doi: 10.1016/0304-4165(67)90109-2. [DOI] [PubMed] [Google Scholar]
- RINEHART J. F., ABUL-HAJ S. K. An improved method for histologic demonstration of acid mucopolysaccharides in tissues. AMA Arch Pathol. 1951 Aug;52(2):189–194. [PubMed] [Google Scholar]
- Rambourg A., Hernandez W., Leblond C. P. Detection of complex carbohydrates in the Golgi apparatus of rat cells. J Cell Biol. 1969 Feb;40(2):395–414. doi: 10.1083/jcb.40.2.395. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- SPICER S. S. DIAMINE METHODS FOR DIFFERENTIALING MUCOSUBSTANCES HISTOCHEMICALLY. J Histochem Cytochem. 1965 Mar;13:211–234. doi: 10.1177/13.3.211. [DOI] [PubMed] [Google Scholar]
- SPICER S. S. Histochemical properties of mucopolysaccharixe and basic protein in mast cells. Ann N Y Acad Sci. 1963 Feb 26;103:322–333. doi: 10.1111/j.1749-6632.1963.tb53707.x. [DOI] [PubMed] [Google Scholar]
- Sampson D., Archer G. T. Release of histamine from human basophils. Blood. 1967 May;29(5):722–736. [PubMed] [Google Scholar]
- Singher H. O., Marinelli L. DISTRIBUTION OF RADIOACTIVE SULFUR IN THE RAT. Science. 1945 Apr 20;101(2625):414–415. doi: 10.1126/science.101.2625.414. [DOI] [PubMed] [Google Scholar]
- Spicer S. S., Staley M. W., Wetzel M. G., Wetzel B. K. Acid mucosubstance and basic protein in mouse Paneth cells. J Histochem Cytochem. 1967 Apr;15(4):225–242. doi: 10.1177/15.4.225. [DOI] [PubMed] [Google Scholar]
- WEISBERGER A. S., SUHRLAND L. G. Comparative incorporation of S35 l-cystine and S35 sodium sulfate by normal and leukemic leukocytes. Blood. 1955 May;10(5):458–466. [PubMed] [Google Scholar]
- Wetzel B. K., Horn R. G., Spicer S. S. Fine structural studies on the development of heterophil, eosinophil, and basophil granulocytes in rabbits. Lab Invest. 1967 Mar;16(3):349–382. [PubMed] [Google Scholar]
- Wetzel B. K., Spicer S. S., Horn R. G. Fine structural localization of acid and alkaline phosphatases in cells of rabbit blood and bone marrow. J Histochem Cytochem. 1967 Jun;15(6):311–334. doi: 10.1177/15.6.311. [DOI] [PubMed] [Google Scholar]
- Wetzel M. G., Wetzel B. K., Spicer S. S. Ultrastructural localization of acid mucosubstances in the mouse colon with iron-containing stains. J Cell Biol. 1966 Aug;30(2):299–315. doi: 10.1083/jcb.30.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Zucker-Franklin D. Electron microscopic study of human basophils. Blood. 1967 Jun;29(6):878–890. [PubMed] [Google Scholar]