Abstract
The fine structure of the nuclear components was studied following mild lysis of mouse or Drosophila tissue culture cells and spreading of nuclear material. Particular attention was paid to nuclear ribonucleoprotein (RNP) constituents, which were analysed by high resolution autoradiography after [3H]uridine pulse labelling of cells. Comparison with the labelling kinetics of various in situ nuclear RNP constituents described previously suggests strong similarities between in situ constituents and structures observed within spread nuclear components. The present observations suggest that the nucleolar dense fibrillar component, shown previously in ultrathin sections of [3H]uridine-labelled intact cells as carrying rapidly labelled pre-rRNA, in fact consists of highly compacted transcribing ribosomal genes. The growing RNP fibrils appearing in transcription complexes of extranucleolar active genes and the in situ observed perichromatin fibrils also show the same labelling properties. This confirms that the two structures indeed represent the same nucleoplasmic constituents. As for the nuclear structures involved in post-transcriptional events, our observations demonstrate the occurrence of a rapidly labelled RNP fibro-granular network. Its granular elements correspond, in size and perichromatin location, to the perichromatin granules seen in in situ preparations and suggest similarities between the two constituents. The results are discussed in the light of other data providing information on the role of various nuclear structural constituents.
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References
Bouvier D, Hubert J, Bouteille M (1980) The nuclear shell in HeLa cell nuclei: whole-mount electron microscopy of the dissociated and isoalted nuclear periphery. J Ultrastruct Res 73:288–298
Bouvier D, Hubert J, Seve AP, Bouteille M (1982) RNA is responsible for the three-dimensional organization of nuclear matrix proteins in HeLa cells. Biol Cell 43:143–146
Dimova RN, Gajdardjieva KC, Dabeva MD, Hadjiolov AA (1979) Early effects of d-galactosamine on rat liver nucleolar structures. Biol Cell 35:1–10
Fakan S (1978) High resolution autoradiography studies on chromatin functions. In: Busch H (ed) The cell nucleus, vol 5. Academic Press, New York, pp 3–53
Fakan S (1986) Structural support for RNA synthesis in the cell nucleus. In: Jasmin G, Simard R (eds) Methods and achievements in experimental pathology, vol 12. Karger S, Basel, pp 105–140
Fakan S, Bernhard W (1971) Localization of rapidly and slowly labeled nuclear RNA as visualized by high resolution autoradiography. Exp Cell Res 67:129–141
Fakan S, Fakan J (1987) Autoradiography of spread molecular complexes. In: Sommerville J, Scheer U (eds) Electron microscopy in molecular biology — A practical approach. IRL Press, Oxford, pp 201–214
Fakan S, Puvion E (1980) The ultrastructural visualization of nucleolar and extranucleolar RNA synthesis and distribution. Int Rev Cytol 65:255–299
Fakan S, Puvion E, Spohr G (1976) Localization and characterization of newly synthesized nuclear RNA in isolated rat hepatocytes. Exp Cell Res 99:155–164
Fakan S, Leser G, Martin TE (1984) Ultrastructural distribution of nuclear ribonucleoproteins as visualized by immunocytochemistry on thin sections. J Cell Biol 98:358–363
Fakan S, Leser G, Martin TE (1986) Immunoelectron microscope visualization of nuclear ribonucleoprotein antigens within spread transcription complexes. J Cell Biol 103:1153–1157
Ghosh S, Paweletz N (1987) Active ribosomal cistrons and their primary transcripts located in the nucleolus. Cell Biol Int Rep 11:205–210
Goessens G (1984) Nucleolar structure. Int Rev Cytol 87:107–158
Granboulan N, Granboulan P (1965) Cytochimie ultrastructurale du nucléole. II Études des sites de synthèse du RNA dans le nucléole et le noyau. Exp Cell Res 38:604–619
Gruca S, Krzyzowska-Gruca S, Vorbrodt A, Krawczyk Z (1978) Intranucleolar localization of the RNA polymerase A activity in isolated nuclei of regenerating liver. Exp Cell Res 114:462–467
Haase G, Jung G (1964) Herstellung von Einkornschichten aus photografischen Emulsionen. Naturwissenschaften 51:404–405
Herman R, Weymouth L, Penman S (1978) Heterogeneous nuclear RNA-protein fibers in chromatin-depleted nuclei. J Cell Biol 78:663–674
Hubert J, Bouvier D, Bouteille M (1979) Nuclear cortex: dissociation of nuclei isolated from cultured human cells. Biol Cell 36:87–90
Hughes ME, Bürki K, Fakan S (1979) Visualization of transcription in early mouse embryos. Chromosoma 73:179–190
Kaufmann SH, Coffey DS, Shaper JH (1981) Considerations in the isolation of rat liver nuclear matrix, nuclear envelope and pore complex lamina. Exp Cell Res 132:105–123
Miller OL, Bakken AH (1972) Morphological studies on transcription. Acta Endocrinol [Suppl] 168:155–173
Miller OL, Beatty BR (1969) Visualization of nucleolar genes. Science 164:955–957
Monneron A, Bernhard W (1969) Fine structural organization of the interphase nucleus in some mammalian cells. J Ultrastruct Res 27:266–288
Müller M, Spiess E, Werner D (1983) Fragmentation of “nuclear matrix” on a mica target. Eur J Cell Biol 31:158–166
Nash RE, Puvion E, Bernhard W (1975) Perichromatin fibrils as components of rapidly labeled extranucleolar RNA. J Ultrastruct Res 53:395–405
Puvion E, Moyne G (1978) Intranuclear migration of newly synthesized extranucleolar ribonucleoprotiens. Exp Cell Res 115:79–88
Puvion E, Moyne G (1981) In situ localization of RNA structures. In: Busch H (ed) The cell nucleus, vol 8. Academic Press, New York, pp 59–115
Puvion-Dutilleul F, Puvion E (1980) New aspects of intranuclear structures following partial decondensation of chromatin: a cytochemical and high resolution autoradiographical study. J Cell Sci 42:305–321
Scheer U, Raska I (1987) Immunocytochemical localization of RNA polymerase I in the fibrillar centers of nucleoli. Chromosomes Today 9:284–294
Scheer U, 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
Stahl A (1982) The nucleolus and nucleolar chromosomes. In: Jordan EG, Cullis CA (eds) The nucleolus. Cambridge University Press, Cambridge, pp 1–24
Thiry M, Lepoint A, Goessens G (1985) Re-evaluation of the site of transcription in Ehrlich tumour cell nucleoli. Biol Cell 54:57–64
Villard D, Fakan S (1978) Visualisation des complexes de transcription dans la chromatine étalée de cellules de mammifères: étude en autoradiographie à haute résolution. C R Acad Sci Ser D 286:777–780
Wisse E, Tates AD (1968) A gold latensification-Elon ascorbic acid developer for Ilford L4 emulsion. Fourth European Regional Conference on Electron Microscopy, Rome, pp 465–466
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Fakan, S., Hughes, M.E. Fine structural ribonucleoprotein components of the cell nucleus visualized after spreading and high resolution autoradiography. Chromosoma 98, 242–249 (1989). https://doi.org/10.1007/BF00327309
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DOI: https://doi.org/10.1007/BF00327309