Summary
-
1.
Nuclei of the different types of granulocytopoietic cells from normal human bone marrow smears were stained with Fastgreen FCF and their dye content was measured cytophotometrically.
-
2.
The very immature cell types (myeloblasts and promyelocytes) showed a high percentage of unstained nuclei or nuclei with a dye content too faint to be measured. More mature stages showed increasing Fastgreen values which were highest in mature polymorphonuclear leukocytes.
-
3.
The changing Fastgreen dye content of nuclei from different cell types—stained specifically for nuclear histone proteins—must be interpreted as varying stainability of histone proteins depending upon the functional state of chromatin. It does not indicate an altering histone content of the nuclei.
Zusammenfassung
-
1.
An Knochenmarksausstrichen eines gesunden Menschen wurde die Fastgreen-Färbbarkeit der Kerne der verschiedenen Granulozytopoesezellen zytophotometrisch untersucht.
-
2.
In den sehr unreifen Myeloblasten und Promyelozyten ist eine Fastgreen-Färbbarkeit der meisten Kerne nicht oder in nicht meßbarer Stärke vorhanden. Am stärksten ist die Fastgreen-Färbbarkeit der Kerne reifer Granulozyten.
-
3.
Die bei verschiedenen Zellarten verschieden starke Anfärbung der Kerne mit Fastgreen bei pH 8,1, die für Histonproteine spezifisch sein soll, wird nicht als Ausdruck einer Histonmengenänderung, sondern als funktionsabhängige Änderung der Fastgreenkapazität der Histonproteine interpretiert.
Similar content being viewed by others
Literatur
Alfert, M.: Changes in the staining capacity of nuclear components during cell degeneration. Biol. Bull. 109, 1–2 (1955).
—: Cytochemische Untersuchungen an basischen Kernproteinen während der Gametenbildung, Befrachtung und Entwicklung. Ges. physiol. chem. Colloq. 9, 73–84 (1958a).
—: Variations in cytochemical properties of cell nuclei. Exp. Cell Res., Suppl. 6, 227–235 (1958b).
—: Geschwind, I. I.: A selective staining for the basic proteins of cell nuclei. Proc. nat. Acad. Sci. (Wash.) 39, 991–999 (1953).
Allfrey, V. G.: Control mechanisms in RNA synthesis. Cancer Res. 26, 2026–2040 (1966).
—: Faulkner, R., Mirsky, A. E.: Acetylation and methylation of histones and their possible role in the regulation of RNA synthesis. Proc. nat. Acad. Sci. (Wash.) 51, 786–794 (1964).
—: Pogo, B. G. T., Littau, V. G., Gershey, E. L., Mirsky, A. E.: Histone acetylation in insect chromosomes. Science 159, 314–316 (1968).
Ansley, H.: A cytophotometric study of chromosome pairing. Chromosoma (Berl.) 8, 380–395 (1957).
Asao, T.: Regional histone changes in embryos of the newt, Trituras pyrrhogaster, during early development. Exp. Cell Res. 61, 255–265 (1970).
Auer, G., Zetterberg, A., Killander, D.: Changes in binding between DNA and arginine residues in histone induced by cell crowding. Exp. Cell Res. 62, 32–38 (1970).
Berlowitz, L., Palotta, D., Pawlowski, P.: Isolated histone fractions and the alkaline fastgreen reaction. J. Histochem. Cytochem. 18, 334–339 (1970).
Black, M. M., Ansley, H. R.: Antigen-induced changes in lymphoid cell histones I. Thymus. J. Cell Biol. 26, 201–208 (1965a).
—: Antigen-induced changes in lymphoid cell histones. II. Regional Lymph-Nodes. J. Cell Biol. 26, 797–803 (1965b).
—: Histone specificity revealed by ammoniacal silver staining. J. Histochem. Cytochem. 14, 177–181 (1966).
—: Antigen-induced changes in lymphoid cell histones. III. In vitro and in Extract. J. Cell Biol. 35, 619–625 (1967).
Bloch, D. P.: Genetic implications of histone bahavior. J. cell. comp. physiol. 62, 87–94 (1963).
Bloch, D. P. Cytochemistry of the histones. Protoplasmatologia V/3d, 1–56 (1966).
—: A microspectrophotometric study of the synthesis of DNA and nuclear histone. J. biophys. biochem. Cytol. 1, 17–28 (1955).
— Hew, H. Y. C.: Changes in nuclear histones during fertilization and early embryonic development in the pulmonate snail Helix aspersa. J. biophys. biochem. Cytol. 8, 69–81 (1960).
Bondy, S. C., Roberts, S., Morelos, B. S.: Histone-acetylating enzyme of brain. Biochem. J. 119, 665–672 (1970).
Burton, D. W.: Initial changes in the deoxyribonucleoprotein complexes of kangaroo lymphocytes stimulated with phytohaemagglutinin. Exp. Cell Res. 49, 330–304 (1968).
Crampton, C. F., Stein, W. H., Moore, S.: Comparative studies on chromatographically purified histones. J. biol. Chem. 225, 363–386 (1957).
Davison, P. F., Butler, J. V.: The chemical composition of calf thymus nucleoprotein. Biochim. biophys. Acta (Amst.) 21, 568–573 (1956).
Deitch, A. D.: Microspectrophotometric studies of the binding of the anionic dye, Naphthol yellow S by tissue sections and by purified proteins. Lab. Invest. 4, 324–351 (1955).
—: A cytophotometric method for the estimation of histone and non-histone protein. J. Histochem. Cytochem. 13, 17–18 (1965).
Frenster, J. H.: Control of DNA strand separation during repression and derepression of DNA synthesis. J. Cell Biol. 27, 30a (1965).
—: A model of specific de-repression within interphase chromatin. Nature (Lond.) 206, 1269–1270 (1965).
Garcia, A. M.: Studies on DNA in leukocytes and related cells of mammals. VII. The Fast Green histone content of rabbit leukocytes after hypotonic treatment. J. Histochem. Cytochem. 17, 475–481 (1969).
Harbers, E.: Zur Rolle der Histone und der sauren Proteine in den Desoxyriboproteinen (Nucleohistonen) des Zellkerns. Dtsch. med. Wschr. 90, 2074–2078 (1965).
Holtzman, E.: A cytochemical study of the solubilities of the histones of fixed Necturus liver. J. Histochem. Cytochem. 13, 318–327 (1965).
Horn, E. C.: Extranuclear histone in the amphibian oocyte. Proc. nat. Acad. Sci. (Wash.) 48, 257–265 (1962).
Kischer, C. W., Gurley, L. R., Shepherd, G. R.: Nuclear histones and early embryogenesis of the chick. Nature (Lond.) 212, 304–306 (1966).
Kleinsmith, L. J., Allfrey, V. G., Mirsky, A. E.: Phosphoprotein metabolism in isolated lymphocyte nuclei. Proc. nat. Acad. Sci (Wash.) 55, 1182–1189 (1966).
—: Phosphorylation of nuclear protein early in the course of gene activation in lymphocytes. Science 154, 780–781 (1966).
Knobloch, A., Matsudaira, H., Vendrely, R.: Étude biochimique comparée des nucléohistones des différentes vertébrées. C. R. Acad. Sci. (Paris) 224, 2980–2983 (1957).
Kruh, J., Tichonicky, L., Dastugue, B.: Histones, protéines acides de la chromatine et biosynthése de l'hémoglobine. Bull. Soc. Chim. biol. (Paris) 52, 1287–1288 (1970).
Langan, T. A.: A phosphoprotein preparation from liver nuclei and its effect on the inhibition of RNA synthesis by histones. In: Regulation of nucleic acid and protein biosynthesis. Ed. by V. V. Koningsberger and L. Bosch (B. B. A. Library, B. 10), S. 233–242. Amsterdam-London-New York: Elsevier Publ. Comp. 1967.
—: Phosphorylation of liver histone following the administration of glucagon and insulin. Proc. nat. Acad. Sci. (Wash.) 64, 1276–1283 (1969).
Lechenault, H.: Étude cytophotométrique des acides nucléiques et des histones des cellules activées au cours de la régénération céphalique de l'Oligochète Eisenia foetida (Sav.) Histochemie 23, 258–366 (1970).
Lillie, R. D.: Histopathologic technic and practical histochemistry, 3rd ed. p. 38. New York-Toronto-Sidney-London: McGraw-Hill Book Co. 1965.
Littau, V. C., Burdick, C. J., Allfrey, V. G., Mirsky, A. E.: The role of histones in the maintenance of chromatin structure. Proc. nat. Acad. Sci. (Wash.) 54, 1204–1212 (1965).
MacRae, E. K., Meetz, G. D.: Electron microscopy of the ammoniacal silver reaction for histones in the erythropoietic cells of the chick. J. Cell Biol. 45, 235–245 (1970).
Moore, B. C.: Histones and differentiation. Proc. nat. Acad. Sci. (Wash.) 50, 1018–1026 (1963).
Noeske, K.: Diskrepanzen von Feulgen-Wert und DNS-Gehalt. Histochemie 20, 322–327 (1969).
—, Ebner, H.: Zum Problem der Hyperchromasie. Verh. Dtsch. Ges. Path. 49, Tagg, 327–330 (1965).
Paik, W. K., Kiu, S.: Enzymatic methylation of histones. Arch. Biochem. 134, 632–637 (1969).
Pogo, B. G. T., Allfrey, V. G., Mirsky, A. E.: RNA synthesis and histone acetylation during the course of gene activation in lymphocytes. Proc. nat. Acad. Sci. (Wash.) 55, 805–812 (1966).
— Pogo, A. O., Allfrey, V. G., Mirsky, A. E.: Changing patterns of histone acetylation and RNA synthesis in regeneration of the liver. Proc. nat. Acad. Sci. (Wash.) 59, 1337–1344 (1968).
Queisser, W., Noeske, K., Sandritter, W., Lennert, K.: Cytophotometrische Untersuchungen des DNS-, Histon-und Gesamtproteingehalts von Epitheloidzellen, Gewebsmastzellen und von Zellen der Plasmocytopoiese bei käsiger Lymphknotentuberkulose. Klin. Wschr. 45, 1135–1142 (1967).
Ruch, F., Rosselet, A.: A cytochemical study of euchromatin and heterochromatin in roots of Rhoeo discolor. Exp. Cell Res. 62, 219–227 (1970).
Rüppell, V., Wolter, J., Sandritter, W.: Cytophotometrische Untersuchungen an Epitheloidzellen verschiedener Lymphknotengranulome. Beitr. path. Anat. 140, 379–406 (1970).
Sandritter, W.: Methods and Results in quantitative cytochemistry In: G. L. Wied: Introduction to quantitative cytochemistry. p. 159–182. New York and London: Academic Press 1966.
—, Mondorf, W., Schiemer, H. G., Müller, D.: Beschreibung eines Zytophotometers für sichtbares Licht. Mikroskopie 14, 25–35 (1959).
Sauter, J. J., Marquardt, H.: Nucleohistone und Ribonucleinsäure-Synthese während der Pollenentwicklung. Naturwissenschaften 54, 546 (1967).
Schwenke, K. D.: Fraktionierung, Struktur und mögliche biologische Funktion der Histone. Z. Chem. 7, 91–101 (1967).
Seitz, J. F.: The biochemistry of the cells of blood and bone marrow, p. 84. Springfield, Ill.: Ch. C. Thomas, Publ. 1969.
Sekeris, C. E., Sekeri, K. E., Gallwitz, D.: The methylation of the histones of rat liver nuclei in vitro. Hoppe-Seylers Z. physiol. Chem. 348, 1660–1666 (1967).
Stocken, L. A.: Zit. in: Biological macromolecules: Interactions in the nucleus. Nature (Lond.) 223, 348–349 (1969).
Takaku, F.. Nakao, K., Ono, T., Terayama, H.: Changes in histone acetylation and RNA synthesis in the spleen of polycythemic mouse after erythropoietin injection. Biochim. biophys. Acta (Amst.) 195, 396–400 (1969).
Tidwell, T., Allfrey, V. G., Mirsky, A. E.: The methylation of histones during regeneration of the liver. J. biol. Chem. 243, 707–715 (1968).
Turner, G., Hancock, R. L.: Histone methylase activity of adult, embryonic and neoplastic liver tissues. Life Sci. 9, II, 917–922 (1970).
Vaughn, J. C.: Changing nuclear histone patterns during development. I. Fertilization and early cleavage in the crab, Emerita analoga. J. Histochem. Cytochem. 16, 473–479 (1968).
Vendrely, R., Alfert, M., Matsudaira, H., Knobloch, A.: The composition of nucleohistone from pycnotic nuclei. Exp. Cell Res. 14, 295–300 (1958).
Wilhelm, J. A., McCarty, K. S.: Partial characterization of the histones and histone acetylation in cell cultures. Cancer Res. 30, 409–417 (1970).
—: The uptake and turnover of acetate in Hela cell histone fractions. Cancer Res. 30, 418–425 (1970).
Zetterberg, A., Auer, G.: Early changes in the binding between DNA and histone in human leukocytes exposed to phythaemagglutinin. Exp. Cell Res. 56, 122–126 (1968).
Author information
Authors and Affiliations
Additional information
Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.
Rights and permissions
About this article
Cite this article
Noeske, K. Stöchiometrische probleme der quantitativen fastgreen-zytophotometrie bei der histonbestimmung. Histochemie 27, 243–252 (1971). https://doi.org/10.1007/BF00264396
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00264396