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The morphological relationship in electron microscopy between NOR-silver proteins and intranucleolar chromatin

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Abstract

The relative distribution of NOR proteins and chromatin fibers in the nucleoli was visualized in human cell line. The chromatin was revealed by a Feulgen-like procedure using osmium-ammine as DNA tracer. This selective staining was combined with NOR-silver staining. We provide morphological evidence for constant overlapping of the silver deposit sites with dispersed intranucleolar chromatin fibers. Silver stained proteins were sometimes observed in contact with the chromatin fibers, suggesting that at least some of the Ag-NOR proteins might be closely connected with the dispersed nucleolar DNA.

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References

  • Bourgeois, C.A., Hernandez-Verdun, D., Hubert, J., Bouteille, M.: Silver staining of NORs in electron microscopy. Exp. Cell Res. 123, 449–452 (1979)

    Google Scholar 

  • Bouteille, M., Hernandez-Verdun, D., Dupuy-Coin, A.M., Bourgeois, C.A.: Nucleoli and Nucleolar-related structures in normal, infected and drug treated cells. In: The nucleolus (G. Jordan and C.A. Cullis, eds), 179–211. Cambridge: University Press 1982

    Google Scholar 

  • Buys, C.H.C.M., Osinga, J., Anders, G.J.P.A.: Age-dependent variability of ribosomal RNA-gene activity in man as determined from frequencies of silver staining nucleolus organizing regions on metaphase chromosomes of lymphocytes and fibroblastes. Mech. Ageing Dev. 11, 55–75 (1979)

    Google Scholar 

  • Buys, C.H.C.M., Osinga, J.: Abundance of protein-bound sulfhydryl and disulfide groups at chromosomal nucleolus organizing regions. Chromosoma (Berl.) 77, 1–11 (1980)

    Google Scholar 

  • Cheng, D.M., Denton, T.E., Liem, S.L., Elliot, C.L.: Variation in nucleolar organizer activity in lymphocytes of females with adenocarcinoma. Clinical Genet. 19, 145–148 (1981)

    Google Scholar 

  • Cogliati, R., Gautier, A.: Mise en évidence de l'ADN et des polysaccharides à l'aide d'un nouveau réactif “de type Schiff”. C.R. Acad. Sci. 276, 3041–3044 (1973)

    Google Scholar 

  • Daskal, Y,, Smetana, K., Busch, H.: Evidence from studies on segregated nucleoli that nucleolar silver staining proteins C23 and B23 are in the fibrillar component. Exp. Cell Res. 127, 285–291 (1980)

    Google Scholar 

  • Derenzini, M.: Fine structure of chromatin as visualized in thin section with the Gautier selective stain for DNA. J. Ultrastruct. Res. 69, 239–248 (1979)

    Google Scholar 

  • Derenzini, M., Hernandez-Verdun, D., Bouteille, M.: Relative distribution of DNA and NOR-proteins in nucleoli visualized by simultaneous Feulgen-like and Ag-NOR staining procedures. Biol. Cell 40, 147–150 (1981a)

    Google Scholar 

  • Derenzini, M., Hernandez-Verdun, D., Bouteille, M.: Subunit configuration of rat hepatocyte chromatin fixed in situ as visualized in thin sections selectively stained for DNA. Biol. cell. 41, 161–163 (1981b)

    Google Scholar 

  • Ellinger von, A., Wachtler, F.: Über eine Methode zur Darstellung des Nucleolus im Licht- und Elektronenmikroskop. Mikroskopie 36, 330–335 (1980)

    Google Scholar 

  • Fakan, S., Puvion, E.: The ultrastructural visualization of nucleolar and extra-nucleolar RNA synthesis and distribution. Int. Rev. Cytol. 65, 255–299 (1980)

    Google Scholar 

  • Gautier, A.: Ultrastructural localization of DNA in ultrathin tissue sections. Int. Rev. Cytol. 44, 113–191 (1976)

    Google Scholar 

  • Goessens, G.: High resolution autoradiographic studies of Ehrlich tumour cell nucleoli. Exp. Cell Res. 100, 88–94 (1976)

    Google Scholar 

  • Goessens, G.: Relations between fibrillar centres and nucleolus-associated chromatin in Ehrlich tumour cells. Cell Biol. Intern. Reports 3, 337–343 (1979)

    Google Scholar 

  • Goessens, G., Lepoint, A.: The nucleolus-organizing regions (NOR's): recent data and hypotheses. Biol. Cell. 35, 211–220 (1979)

    Google Scholar 

  • Goodpasture, C., Bloom, S.E.: Visualization of nucleolar organizer regions in mammalian chromosome using silver staining. Chromosoma (Berl.) 53, 37–50 (1975)

    Google Scholar 

  • Hansmann, I., Gebauer, J., Bihl, L., Grimm, T.: Onset of nucleolus organizer activity in early mouse embryogenesis and evidence for its regulation. Exp. Cell Res. 114, 263–268 (1978)

    Google Scholar 

  • Hartung, M., Mirre, C., Stahl, A.: Nucleolar organizers in human oocytes at meiotic prophase I, studied by the silver-NOR method and electron microscopy. Hum. Genet. 52, 295–308 (1979)

    Google Scholar 

  • Heneen, W.K.: Silver staining of nucleolar organizers and nucleoli during mitosis and interphase of mammalian cells. Genetica 50, 31–39 (1979)

    Google Scholar 

  • Hens, L., Kirsch-Volders, M., Arrighi, F.E., Susanne, C.: Relationship between measured chromosome distribution parameters and Ag-staining of the nucleolus organizer regions. Hum. Genet. 53, 363–370 (1980)

    Google Scholar 

  • Hernandez-Verdun, D., Bouteille, M.: Nucleologenesis in chick erythrocyte nuclei reactivated by cell fusion. J. Ultrastr. Res. 69, 164–179 (1979)

    Google Scholar 

  • Hernandez-Verdun, D., Hubert, J., Bourgeois, C.A., Bouteille, M.: Identification ultrastructurale de l'organisateur nucléolaire par la technique à l'argent. C.R. Acad. Sc. Paris 287, 1421–1423 (1978)

    Google Scholar 

  • Hernandez-Verdun, D., Hubert, J., Bourgeois, C.A., Bouteille, M.: Ultrastructural localization of Ag-NOR stained proteins in the nucleolus during the cell cycle and in other nucleolar structures. Chromosoma (Berl.) 79, 349–362 (1980)

    Google Scholar 

  • Hofgärtner, F.J., Schmid, M., Krone, W., Zenzes, M.T., Engel, W.: Pattern of activity of nucleolus organizers during spermatogenesis in mammals as analyzed by silverstaining. Chromosoma (Berl.) 71, 197–216 (1979a)

    Google Scholar 

  • Hofgärtner, F.J., Krone, W., Jain, K.: Correlated inhibition of ribosomal RNA synthesis and silver staining by actinomycin D. Hum. Genet. 47, 329–333 (1979b)

    Google Scholar 

  • Hsu, T.C., Spirito, S.E., Pardue, M.L.: Distribution of 18 + 28S ribosomal genes in mammalian genomes. Chromosoma (Berl.) 53, 25–36 (1975)

    Google Scholar 

  • Hubbell, H.R., Hsu, T.C.: Identification of nucleolus organizer regions (NORs) in normal and neoplastic human cells by the silver-staining technique. Cytogenet. Cell Genet. 19, 185–196 (1977)

    Google Scholar 

  • Hubbell, H.R., Rothblum, L.I., Hsu, T.C.: Identification of a silver binding protein associated with the cytological silver staining of actively transcribing nucleolar regions. Cell Biol. Internat. Reports 3, 615–622 (1979)

    Google Scholar 

  • Hubbell, H.R., Yun-Fai, Lau., Brown, R.L., Hsu, T.C.: Cell cycle analysis and drug inhibition studies of silver staining in synchronous Hela cells. Exp. Cell Res. 129, 139–147 (1980)

    Google Scholar 

  • Lischwe, M.A., Smetana, K., Olson, M.O.J., Busch, H.: Proteins C23 and B23 are the major nucleolar silver staining proteins. Life Sciences 25, 701–708 (1979)

    Google Scholar 

  • Miller, O.J., Miller, D.A., Dev, V.G., Tantravahi, R., Croce, C.M.: Expression of human and suppression of mouse nucleolus organizer activity in mouse — human somatic cell hybrids. Proc. Natl. Acad. Sci. 73, 4531–4535 (1976a)

    Google Scholar 

  • Miller, D.A., Dev, V.G., Tantravahi, R., Miller, O.J.: Suppression of human nucleolus organizer activity in mouse — human somatic hybrid cells. Exp. Cell Res. 101, 235–243 (1976b)

    Google Scholar 

  • Mirre, C., Stahl, A.: Peripheral RNA synthesis of fibrillar center in nucleoli of japanese quail oocytes and somatic cells. J. Ultrastruct. Res. 64, 377–387 (1978)

    Google Scholar 

  • Mirre, C., Stahl, A.: Ultrastructural organization, sites of transcription and distribution of fibrillar centers in the nucleolus of the mouse oocyte. J. Cell Sci. 48, 105–126 (1981)

    Google Scholar 

  • Moyne, G.: Methods in ultrastructural cytochemistry of the cell nucleus. Prog. Histochem. Cytochem. 13, 1–72 (1980)

    Google Scholar 

  • Olert, J., Sawatzki, G., Kling, H., Gebauer, J.: Cytological and histochemical studies on the mechanism of the selective silver staining of nucleolus organizer regions (NORs). Histochemistry 60, 91–99 (1979)

    Google Scholar 

  • Pebusque, M.J., Vio, M., Seite, R.: Ultrastructural location of Ag-NOR stained proteins in nucleoli of rat sympathetic neurons during the dark period. Biol. Cell 40, 151–154 (1981)

    Google Scholar 

  • Pelliccia, F., Capoa, A. de, Belloni, G., Rocchi, A., Ferraro, M.: Localization of silver staining in interphase, prophase and metaphase lymphocytes. Exp. Cell Res. 115, 439–441 (1978)

    Google Scholar 

  • Ray, M., Pearson, J.: Nucleolar organizing regions of human chromosomes. Hum. Genet. 48, 201–210 (1979)

    Google Scholar 

  • Raynal, F., Amalric, F., Bachellerie, J.P., Puvion-Dutilleul, F., Zalta, J.P.: Integrity of ribosomal transcription complexes after isolation of nucleoli from chinese hamster ovary cells. Biol. Cell 41, 29–34 (1981)

    Google Scholar 

  • Scheer, U.: Changes of nucleosome frequency in nucleolar and non-nucleolar chromatin as a function of transcription: an electron microscopic study. Cell 13, 535–549 (1978)

    Google Scholar 

  • Scheer, U.: Structural organization of spacer chromatin between transcribed ribosomal RNA genes in amphibian oocytes. Europ. J. Cell Biol. 23, 189–196 (1980)

    Google Scholar 

  • Scheer, U., Trendelenburg, M.F., Franke, W.W.: Regulation of transcription of ribosomal RNA-genes during amphibian oogenesis. Progress Different. Res. (Müller-Berat et al. eds, North-Holland Publishing Company, Amsterdam, 105–118 (1976)

    Google Scholar 

  • Schmiady, H., Münke, M., Sperling, K.: Ag-staining of nucleolus organizer regions on human prematurely condensed chromosomes from cells with different ribosomal RNA gene activity. Exp. Cell Res. 121, 425–428 (1979)

    Google Scholar 

  • Schwarzacher, H.G., Mikelsaar, A-V., Schnedl, W.: The nature of the Ag-staining of nucleolus organizer regions. Electron and light-microscopic studies on human cells in interphase, mitosis and meiosis. Cytogen. Cell Genet. 20, 24–39 (1978)

    Google Scholar 

  • Tantravahi, R., Miller, D.A., Dev, V.G., Miller, O.J.: Detection of nucleolus organizer regions in chromosomes of Human, Chimpanzee, Gorilla, Orangutan and Gibbon. Chromosoma (Berl.) 56, 15–27 (1976)

    Google Scholar 

  • Wachtler, F., Ellinger, A., Schwarzacher, H.G.: Nucleolar changes in human phytohaemagglutinin-stimulated lymphocytes. Cell Tissue Res. 213, 351–360 (1980)

    Google Scholar 

  • Williams, M.A., Kleinschmidt, J.A., Krohne, G., Franke, W.W.: Argyrophilic nuclear and nucleolar proteins of Xenopus laevis oocytes identified by gel electrophoresis. Exp. Cell Res. 137, 341–351 (1982)

    Google Scholar 

  • Zankl, H., Mayer, C., Zang, K.D.: Association frequency and silver staining of nucleolus organizing regions in hyperthyroid patients. Hum. Genet. 54, 111–114 (1980)

    Google Scholar 

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Authors and Affiliations

  1. Laboratoire de Pathologie cellulaire, 15 rue de l'Ecole de Médecine, 75270, Paris Cedex 06, France

    D. Hernandez-Verdun, M. Derenzini & M. Bouteille

  2. Istituto di Patologia generale dell'Università di Bologna, via S. Giacomo 14, 40126, Bologna, Italy

    M. Derenzini

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  1. D. Hernandez-Verdun
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  2. M. Derenzini
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  3. M. Bouteille
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Hernandez-Verdun, D., Derenzini, M. & Bouteille, M. The morphological relationship in electron microscopy between NOR-silver proteins and intranucleolar chromatin. Chromosoma 85, 461–473 (1982). https://doi.org/10.1007/BF00327343

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  • DOI: https://doi.org/10.1007/BF00327343

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