Summary
In embryonic cell-line derivative KCo of Drosophila melanogaster, the nucleolus, like most nucleoli, contains a small proportion of ribosomal DNA (1–2% of the total nucleolar DNA). The ribosomal DNA is virtually the only active gene set in the nucleolus and is found among long stretches of inactive supercoiled heterochromatic segments. We have demonstrated by use of a Feulgen-like ammine-osmium staining procedure that, depending on the state of growth, more or less fibres of decondensed DNA emanating from the intra-nucleolar chromatin (which is in continuity with the nucleolus-associated chromatin) ramify and unravel within the central nucleolar core to be transcribed. The nucleolus expands or contracts with the variation of activity and could belong to a supramolecular matricial structure such as is shown after extraction of the nuclei. After a long period of exposure to high doses of actinomycin D, the central nucleolar core became an homogeneous fibrous structure that could be interpreted as an aggregate of protein skeletal elements. The mechanism of repression and derepression of the nucleolar chromatin could thus be explained by a mechanism involving in part a sub-nucleolar structure. We propose a schematic organization of the nucleolar chromatin in KCo cells of Drosophila and discuss it in relation with other nucleolar organizations.
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References
Berezney R (1979) Dynamic properties of the nuclear matrix. In: Busch H (ed) The cell nucleus. Academic-Press, New York, vol 7, pp 413–456
Berezney R, Buchholtz LA (1981) Dynamic association of replicating DNA fragments with the nuclear matrix of regenerating liver. Exp Cell Res 132:1–13
Berezney R, Macaulay LK, Crane FL (1972) The purification and biochemical characterization of bovine liver nuclear membranes. J Biol Cell 247:5549–5561
Bernhard W (1969) A new staining procedure for electron microscopical cytology. J Ultrastruc Res 27:250–265
Bernhard W (1971) Drug induced changes in the interphase nucleus. In: Clementi F, Ceccarelli B (ed). Advances in cytopharmacology. Raven-Press, New York, vol 1, pp 49–67
Brasch K (1982) Fine structure and localization of the nuclear matrix in situ. Exp Cell Res 140:161–171
Burton K (1956) A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of DNA. Biochem J 62:315–321
Busch H, Smetana K (1970) The nucleolus Academic Press, New York
Cogliati R, Gautier A (1973) Mise en évidence de l'ADN et des polysaccharides à l'aide d'un nouveau réactif de type “Schiff”. Cr Hebd Seanc Acad Sci Paris 276:3041–3044
Derenzini M, Viron A, Puvion-Dutilleul F (1982a) The Feulgenlike osmium-ammine reaction as a tool to investigate chromatin structure in thin sections. J Ultrastruct Res 80:133–147
Derenzini M, Hernandez-Verdun D, Bouteille M (1982b) Visualization in situ of extended DNA filaments in nucleolar chromatin of rat hepatocytes. Exp Cell Res 141:463–468
Echallier G, Ohanessian A (1970) In vitro culture of Drosophila melanogaster embryonic cells. In Vitro 6:162–172
Fakan S, Puvion E (1980) The ultrastructural visualization of nucleolar and extranucleolar RNA synthesis and distribution. Int Rev Cytol 65:255–269
Franke WW, Kleinschmidt JA, Spring H, Krohne G, Grund C, Trendelenburg MF, Stoehr M, Scheer U (1981) A nucleolar skeleton of protein filaments demonstrated in amplified nucleoli of Xenopus laevis. J Cell Biol 90:285–299
Gillespie D, Spiegelman S (1965) A quantitative assay for RNA/ DNA hybrids with DNA immobilized on a membrane. J Mol Biol 12:829–842
Goessens G, Lepoint A (1979) The nucleolus-organizing regions (NORs): Recent data and hypothesis. Biol Cell 35:211–220
Gosh S, Paweletz N, Gosh I (1978) Cytological identification and characterization of the nuclear matrix. Exp Cell Res 111:363–371
Herlan G, Quevedo R, Wunderlich F (1978) Structural transformation of the nuclear matrix in situ. Exp Cell Res 115:103–110
Jackson DA, McCready SJ, Cook PR (1981) RNA synthesized at the nuclear cage. Nature 292:552–555
Jordan EG, McGovern JH (1982) The quantitative relationship of the fibrillar centres and other nucleolar components to changes in growth conditions, serum deprivation and low doses of actino mycin D in cultured diploid human fibroblasts. J Cell Sci 52:373–389
Kaufmann SH, Coffey DS, Shaper JH (1981) Considerations in the isolation of rat liver nuclear matrix, nucleolar envelope and pore lamina complex. Exp Cell Res 132:105–113
Knibiehler B, Mirre C, Rosset R (1982a) Nucleolar organizer structure and activity in a nucleolus without fibrillar centres: The nucleolus in an established Drosophila cell line. J Cell Sci 57:351–364
Knibiehler B, Bernadac A, Mirre C, Rosset R (1982b) Pattern of RNA synthesis and morphological changes induced at the nucleolar level by ecdysone treatment in a Drosophila KC cell line derivative. Biol Cell 46: in press
Locke M, Huie P (1980) The nucleolus during epidermal development in an insect. Tissue Cell 12:175–194
Mentré P (1972) Coloration sur coupes ultra-fines des acides nucléiques et de certains polysaccharides par le thallium en solution alcoolique. J Microsc 14:251–270
Merriam RW (1969) Movement of cytoplasmic proteins into nuclei induced to enlarge and initiate DNA or RNA synthesis. J Cell Sci 5:333–349
Mirre C, Knibiehler B (1981) Ultrastructural autoradiographic localization of the rRNA transcription sites in the quail nucleolar components using two RNA antimetabolites. Biol Cell 42:73–78
Mirre C, Knibiehler B (1982) A re-evaluation of the relationships between the fibrillar centres and the nucleolus-organizing-regions in reticulated nucleoli: ultrastructural organization, number and distribution of the fibrillar centres in the nucleolus of the mouse Sertoli cell. J Cell Sci 55:247–259
Mirre C, Stahl A (1978) Peripheral RNA synthesis of fibrillar centres in nucleoli of Japanese quail oocytes and somatic cells. J Ultrastruct Res 64:377–387
Mirre C, Stahl A (1981) Ultrastructural organization, sites of transcription and distribution of fibrillar centres in the nucleolus of the mouse oocyte. J Cell Sci 48:105–126
Recher L, Whitescarver J, Briggs LG (1969) Fine ultrastructure of a nucleolar constituent. J Ultrastruct Res 29:1–14
Ritossa MF, Spiegelman S (1965) Localization of DNA complementary to rRNA in the nucleolus organizer region of Drosophila melanogaster. Proc Natl Acad Sci USA 53:737–745
Rodman TC (1969) Morphology and replication of intranucleolar DNA in polytene nuclei. J Cell Biol 42:575–582
Todorov IT, Hadjiolov AA (1979) A comparison of nuclear and nucleolar matrix proteins from rat liver. Cell Biol Int Rep 3:753–757
Unuma T, Senda R, Muramatsu M (1972) Mechanism of nucleolar segregation. Differences in effects of Actinomycin D and Cycloheximide on nucleoli of rat liver cells. J Electron Microsc 22:205–216
Van Eekelen CAG, Salden MHL, Habets WJA, Van de Putte LBA, Van Vendrooij WJ (1982) On the existence of an internal nuclear protein structure in HeLa cells. Exp Cell Res 141:181–190
Wachtler F, Ellinger A, Schwarzacher HG (1980) Nucleolar changes in human phytohaemagglutinin stimulated lymphocytes. Cell Tissue Res 213:351–360
Wunderlich F, Berezney R, Kleinig H (1976) The nuclear envelope: an interdisciplinary analysis of its morphology, composition and functions. In: Chapman D, Wallach DFH (eds). Biological membranes. Academic-Press, New York, vol 3, pp 241–333
Zalta J, Zalta J-P (1973) Basic principles of a method of nucleoli isolation. In: Prescott DM (ed) Methods in cell biology. Academic-Press, New York, vol 6, pp 317–330
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Knibiehler, B., Mirre, C., Navarro, A. et al. Studies on chromatin organization in a nucleolus without fibrillar centres. Cell Tissue Res. 236, 279–288 (1984). https://doi.org/10.1007/BF00214228
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DOI: https://doi.org/10.1007/BF00214228