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Intranucleolar visualization of nucleic acids and acidic proteins in inhibited and reactivated pea cotyledonary buds

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Summary

Nuclease-colloidal gold complexes and silver staining were used to visualize intranucleolar nucleic acids and argyrophilic proteins of the nucleolar organizers in bud cotyledonary cells ofPisum sativum. In the G0–1 inhibited bud, a few RNA molecules were detected in the fibrillar component and in the unique fibrillar centre, close to the boundary with the fibrillar component of the nucleolus. DNA was present in the fibrillar component, in the fibrillar centre and in a few fibres crossing the perinucleolar halo. The acidic proteins were localized at the periphery of the fibrillar component but they were also present in the unique fibrillar centre. In the reactivated bud, RNA was particularly concentrated in the granular component and along fibres crossing the perinucleolar halo; a few RNA molecules were also detected at the boundary between the small fibrillar centres and the fibrillar component. DNA was localized in the same nucleolar component as in the inhibited bud, but it was distributed between several fibrillar centres. Acidic proteins coated these DNA loci. In the inhibited and reactivated bud connections between nucleolar DNA containing structures were displayed. The data are discussed in relation to the present knowledge of the functional architecture of the nucleolus.

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Abbreviations

DNA:

deoxyribonucleic acid

DNase:

deoxyribonuclease

G0–1 :

phase G1 phase of the cell cycle indefinitely prolonged

PEG:

polyethylene glycol

RNA:

ribonucleic acid

RNase:

ribonuclease

S and G2 :

phases synthetic and postsynthetic phases of the cell cycle

SPB:

saline phosphate buffer

References

  • Bendayan M (1981) Electron microscopical localization of nucleic acids by means of nuclease-gold complex. Histochem J 13: 699–710

    PubMed  Google Scholar 

  • — (1982) Ultrastructural localization of nucleic acids by the use of enzyme-gold complexes. Influence of fixation and embedding. Biol Cell 43: 153–156

    Google Scholar 

  • — (1984) Enzyme-gold electron microscopic cytochemistry: a new affinity approach for the ultrastructural localization of macromolecules. J Electron Microsc Tech 1: 349–372

    Google Scholar 

  • —, Stephens H (1984) Double labelling cytochemistry applying the protein A-gold technique. In: Polak JM, Varndell IM (eds) Immunolabelling for electron microscopy. Elsevier, Amsterdam, pp 143–154

    Google Scholar 

  • Cataldo C, Soucher C, Stahl A (1988) Three-dimensional ultrastructure and quantitative analysis of the human Sertoli cell nucleolus. Biol Cell 63: 277–285

    PubMed  Google Scholar 

  • Derenzini M, Hernandez-Verdun D, Bouteille M (1982 a) Visualization in situ of extended DNA filaments in nucleolar chromatin of rat hepatocytes. Exp Cell Res 141: 463–469

    PubMed  Google Scholar 

  • —, Viron A, Puvion-Dutilleul F (1982 b) The Feulgen-like osmiumamine reaction as a tool to investigate chromatin structure in thin sections. J Ultrastruct Res 80: 133–147

    PubMed  Google Scholar 

  • Devictor M, Hartung M, Stahl A (1987) Distribution of rDNA and of its transcription sites in the nucleolus of the human Sertoli cell. In: Stahl A (ed) Chromosomes today, vol 9. Allen & Unwin, London, pp 295–300

    Google Scholar 

  • Frens G (1973) Controlled nucleation for the regulation of the particle size in monodispersed gold suspensions. Nature Phys Sci 241: 20–22

    Google Scholar 

  • Goessens G (1973) Les centres fibrillaires des nucléoles des cellules tumorales d'Ehrlich. CR Acad Sci 277: 325–327

    Google Scholar 

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

    PubMed  Google Scholar 

  • Goodpasture C, Bloom SE (1975) Visualization of nucleolar organizer regions in mammalian chromosomes during silver staining. Chromosoma 53: 37–50

    PubMed  Google Scholar 

  • Howell WM, Denton TE, Diamond JR (1975) Differential staining of the satellite regions of human acrocentre chromosomes. Experientia 31: 260–262

    PubMed  Google Scholar 

  • Jennane A (1989) Réactivation du bourgeon cotylédonaire du pois: nucléole marqueur de reprise du cycle cellulaire. Thèse de Doctorat Université PM Curie, Paris, pp 166

  • —, Landré P, Nougarède A (1987) Nucleolus activation in pea cotyledonary buds during 24 hours after decapitation of the main stem: cytochemical studies. Protoplasma 136: 29–36

    Google Scholar 

  • Lischwe MA, Richards RL, Busch RK, Busch H (1981) Localization of phosphoprotein C23 to nucleolar structures and to the nucleolus organizer regions. Exp Cell Res 136: 101–109

    PubMed  Google Scholar 

  • Mamrack MD, Olson MOJ, Busch H (1979) Amino acid sequence and sites of phosphorylation in a highly acidic region of nucleolar non histone protein C23. Biochemistry 18: 3381–3386

    PubMed  Google Scholar 

  • Medina FJ, Risueño MC, Sanchez-Pina MA, Fernández-Gómez ME (1983) A study on nucleolar silver staining in plant cells. The role of argyrophilic proteins in nucleolar physiology. Chromosoma 88: 149–155

    Google Scholar 

  • —, Solanilla EL, Sanchez-Pina A, Fernández-Gómez ME, Risueño MC (1986) Cytological approach to the nucleolar functions detected by silver staining. Chromosoma 94: 259–266

    Google Scholar 

  • Mirre C, Stahl A (1978) Ultrastructure and activity of the nuclear organizer in the mouse oocyte during meiotic prophase. J Cell Sci 31: 79–100

    PubMed  Google Scholar 

  • Nougarède A (1977) Infrastructure des axillaires cotylédonnaires duPisum sativum L. (var. nain hâtif d'Annonay) durant le blocage en phase G1 (état inhibé) et après la reprise d'activité. CR Acad Sci [D] 284: 25–28

    Google Scholar 

  • —, Rembur J, Rondet P (1981) Réactivation du bourgeon cotylédonaire du pois en réponse à la kinétine. Can J Bot 59: 590–603

    Google Scholar 

  • —, Rondet P, Rembur J (1987) Effet d'un traitement par l'acide abscissique sur la division cellulaire, les teneurs en ADN et l'élongation du bourgeon cotylédonaire des plants de pois décapités. Can J Bot 65: 907–915

    Google Scholar 

  • Ploton D, Bobichon H, Adnet JJ (1982) Ultrastructural localization of NOR in nucleoli of human breast cancer tissues using a one-step Ag-NOR staining method. Biol Cell 43: 229–232

    Google Scholar 

  • —, Ménager M, Adnet JJ (1984) Simultaneous high resolution localization of Ag-NOR proteins and nucleoproteins in interphasic and mitotic nuclei. Histochem J 16: 897–906

    PubMed  Google Scholar 

  • Reynolds S (1963) The use of lead citrate at high pH as an electron opaque stain in electron microscopy. J Cell Biol 17: 208–213

    PubMed  Google Scholar 

  • Risueño MC, Medina FJ, Moreno-Diaz De La Espina S (1982) Nucleolar fibrillar centres in plant meristematic cells: ultrastructure, cytochemistry and autoradiography. J Cell Sci 58: 313–329

    PubMed  Google Scholar 

  • Scheer U, Raska I (1987) Immunocytochemical localization of RNA polymerase I in the fibrillar centers of nucleoli. In: Stahl A (ed) Chromosomes today, vol 9. Allen & Unwin, London, pp 284–294

    Google Scholar 

  • —, Rose KM (1984) Localization of RNA polymerase I in interphase cells and mitotic chromosomes by light and electron microscopic immunocytochemistry. Proc Natl Acad Sci USA 81: 1431–1435

    PubMed  Google Scholar 

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

    PubMed  Google Scholar 

  • Thiry M (1988 a) Study of RNA distribution in the nucleolar components of Ehrlich cell using RNase-gold method. Histochemistry 89: 231–236

    PubMed  Google Scholar 

  • — (1988 b) Immunoelectron microscope localization of bromodeoxyuridine incorporated into DNA of Ehrlich tumor cell nucleoli. Exp Cell Res 179: 204–213

    PubMed  Google Scholar 

  • —, Thiry-Blaise L (1989) In situ hybridization at the electron microscope level: an improved method for precise localization of ribosomal DNA and RNA. Eur J Cell Biol 50: 235–243

    PubMed  Google Scholar 

  • Wassef M, Burglen J, Bernhard W (1979) A new method for visualization preribosomal granules in the nucleolus after acetylation. Biol Cell 34: 153–158

    Google Scholar 

  • Wolff G, Quednau HD (1988) The arrangement ofPisum chromosomes in interphase. Genome 30: 717–722

    Google Scholar 

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Authors and Affiliations

  1. Laboratoire de Cytologie expérimentale et Morphogenèse végétale, Université Pierre et Marie Curie, Bât. N 2, Paris

    Arlette Nougarède, P. Landré & Abdelilah Jennane

Authors
  1. Arlette Nougarède
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  2. P. Landré
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  3. Abdelilah Jennane
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Nougarède, A., Landré, P. & Jennane, A. Intranucleolar visualization of nucleic acids and acidic proteins in inhibited and reactivated pea cotyledonary buds. Protoplasma 156, 183–191 (1990). https://doi.org/10.1007/BF01560656

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  • DOI: https://doi.org/10.1007/BF01560656

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