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Cytochemical and immunocytochemical study of coiled bodies in different cultured cell lines

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Abstract

We analyzed by different cytochemical and immunocytochemical approaches the biochemical compositon of coiled bodies in three different cultured cell lines. Coiled bodies are stained by the AgNOR staining method and by the EDTA regressive staining method preferential for ribonucleoprotein (RNP). Using the in situ polyadenylate nucleotidyl transferase-immunogold technique or anti-RNA antibodies, we decisively demonstrated the presence of appreciable amounts of RNA in coiled bodies. Neither the in situ terminal deoxynucleotidyl transferase-immunogold technique nor anti-DNA antibodies revealed any DNA in coiled bodies. Coiled bodies thus appear as distinct regions of cell nuclei involved in some steps of RNA metabolism but not directly in RNA synthesis. Their relationships with the dense fibrillar component of the nucleolus and with interchromatin granule clusters are discussed.

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References

  • Andrade L, Chan EK, Raska I, Peebles C, Roos G, Tan EM (1991) Humanautoantibody to a novel protein of the nuclear coiled body: Immunological characterization and cDNA cloning of p80-coilin. J Exp Med 173: 1407–1419

    Google Scholar 

  • Andrade L, Tan EM, Chan EKL (1993) Immunocytochemical analysis of the coiled body in the cell cycle and during cell proliferation. Proc Natl Acad Sci USA 90: 1947–1951

    Google Scholar 

  • Baserga S, Yang X, Steitz JA (1991) An intact Box C sequence in the U3 snRNA is required for binding of fibrillarin, the protein common to the major family of nucleolar snRNPs. EMBO J 10: 2645–2651

    Google Scholar 

  • Bendayan M, Stephens H (1984) Double labelling cytochemistry applying the protein A-gold technique. In: Polak J, Varndell I (eds) Immunolabelling of electron microscopy. Elsevier, Amsterdam North-Holland New York, USA, pp 143–154

    Google Scholar 

  • Bernhard W (1969) A new staining procedure for electron microscopical cytology. J Ultrastruct Res 27: 250–265

    Google Scholar 

  • Brasch K, Ochs R (1992) Nuclear bodies: A newly rediscovered organelle. Exp Cell Res 202: 211–223

    Google Scholar 

  • Carmo-Fonseca M, Pepperkok R, Carvalmo MT, Lamond AI (1992) Transcription-dependent colocalization of the U1, U2, U4/U6, and U5 snRNPs in coiled bodies. J Cell Biol 117: 1–14

    Google Scholar 

  • Carmo-Fonseca M, Ferreira J, Lamond AI (1993) Assembly of snRNP-containing coiled bodies is regulated in interphase and mitosis — Evidence that the coiled body is a kinetic nuclear structure. J Cell Biol 120: 841–852

    Google Scholar 

  • Craigh SW, Pollard TD (1982) Actin-binding proteins. Trends Biochem Sci 7: 88–92

    Google Scholar 

  • Eilat D, Fischel R (1991) Recurrent utilization of genetic elements in v regions of antinucleic acid antibodies from autoimmune mice. J Immunol 147: 361–368

    Google Scholar 

  • Eilat D, Webster DM, Rees AR (1988) V region sequences of anti-DNA and anti-RNA autoantibodies from NZB/NZW F1 mice. J Immunol 141: 1745–1753

    Google Scholar 

  • Fakan S, Leser G, Martin T (1984) Ultrastructural distribution of nuclear ribonucleoproteins as visualized by immunocytochemistry on thin sections. J Cell Biol 98: 358–363

    Google Scholar 

  • Fu XD, Maniatis T (1990) Factor required for mammalian spliceosome assembly is localized to discrete regions in the nucleus. Nature 343: 437–441

    Google Scholar 

  • Hardin J, Spicer S, Greene WB (1969) The paranucleolar structure, accessory body of Cajal, sex chromatin, and related structures in nuclei of rat trigeminal neurons: A cytochemical and ultrastructural study. Anat Rec 164: 403–432

    Google Scholar 

  • Huang S, Spector DL (1992) U1 and U2 snRNAs are present in nuclear speckles. Proc Natl Acad Sci USA 89: 305–308

    Google Scholar 

  • Lafarga M, Hervas JP (1983) Light and electron microscopic characterization of the “Accessory Body” of Cajal in the neuronal nucleus. In: Grisolia S, Guerri C, Samson F, Norton S, Reinoso-Suorez F (eds) Ramon y Cajal's Contribution of the Neurosciences. Elsevier, Amsterdam North-Holland, pp 91–100

    Google Scholar 

  • Lafarga M, Hervas J, Santa-Cruz M, Villegas J, Crespo D (1983) The “accessory body” of Cajal in the neuronal nucleus. A light and electron microscopic approach. Anat Embryol 166: 19–30

    Google Scholar 

  • Lafarga M, Andres M, Berciano M, Maquiera E (1991) Organization of nucleoli and nuclear bodies in osmotically stimulated supraoptic neurons of the rat. J Comp Neurol 308: 329–339

    Google Scholar 

  • Lamond A, Carmo-Fonseca M (1993) The coiled body. Trends Cell Biol 3: 198–204

    Google Scholar 

  • Lepoint A, Bassleer R (1973) Etude cytologique et cytochimique de cellules tumorales d'Ehrlich de la souris cultivées in vitro. CR Acad Sci Paris 276: 441–443

    Google Scholar 

  • Matera AG, Ward DC (1993) Nucleoplasmic organization of small nuclear ribonucleoproteins in cultures human cells. J Cell Biol 121: 715–727

    Google Scholar 

  • Monneron A, Bernhard W (1969) Fine structural organization of the interphase nucleus in some mammalian cells. J Ultrastruct Res 27: 266–288

    Google Scholar 

  • Moreno Diaz de la Espina S, Risueno M, Medina F (1982) Ultrastructural, cytochemical and autoradiographic characterization of coiled bodies in the plant cell nucleus. Biol Cell 44: 229–238

    Google Scholar 

  • Ploton D, 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

    Google Scholar 

  • Ramon y Cajal SR (1903) Um sencillo metodo de coloracion seletiva del reticulo protoplasmico y sus efectos en los diversos organos nerviosos de vertebrados y invertebrados. Trab Lab Invest Biol Madrid 2: 129–221

    Google Scholar 

  • Raska I, Ochs R, Andrade L, Chan EK, Burlingame R, Peebles C, Gruol D, Tan EM (1990) Association between the nucleolus and the coiled body. J Struct Biol 104: 120–127

    Google Scholar 

  • Raska I, Andrade L, Ochs R, Chan EK, Chang CM, Roos G, Tan EM (1991) Immunological and ultrastructural studies of the nuclear coiled body with autoimmune antibodies. Exp Cell Res 195: 27–37

    Google Scholar 

  • Recher L, Parry N, Whitescarver J, Briggs L (1972) Lead-positive nuclear structures and their behavior under the effect of various drugs. J Ultrastruct Res 38: 398–410

    Google Scholar 

  • Roth J, Bendayan M, Carlemalm E, Villiger W, Garavito M (1981) Enhancement of structural preservation and immunocytochemical staining in low temperature embedded pancreatic tissue. J Histochem Cytochem 29: 663–671

    Google Scholar 

  • Scheer U, Messner K, Hazan R, Raska I, Hansmann P, Falk H, Spiess E, Franke W (1987) High sensitivity immunolocalization of double- and single-stranded DNA by a monoclonal antibody. Eur J Cell Biol 43: 358–371

    Google Scholar 

  • Seite R, Pebusque MJ, Vio-Cigna M (1982) Argyrophilic proteins on coiled bodies in sympathetic neurons identified by Ag-NOR procedure. Biol Cell 46: 97–100

    Google Scholar 

  • Spector D, Fu XD, Maniatis T (1991) Associations between distinct pre-mRNA splicing components and the cell nucleus. EMBO J 10: 3467–3481

    Google Scholar 

  • Thiry M (1992a) Ultrastructural detection of DNA within the nucleolus by sensitive molecular immunocytochemistry. Exp Cell Res 200: 135–144

    Google Scholar 

  • Thiry M (1992b) Highly sensitive immunodetection of DNA on sections with exogenous terminal deoxynucleotidyl transferase and non-isotopic nucleotide analogues. J Histochem Cytochem 40: 411–419

    Google Scholar 

  • Thiry M (1993a) Ultrastructural distribution of DNA and RNA within the nucleolus of human Sertoli cells as seen by molecular immunocytochemistry. J Cell Sci 105: 33–39

    Google Scholar 

  • Thiry M (1993b) Immunodetection of RNA on ultrathin sections incubated with polyadenylate nucleotidyl transferase. J Histochem Cytochem 41: 657–665

    Google Scholar 

  • Thiry M. Scheer U, Goessens G (1988) Localization of DNA within Ehrlich tumour cell nucleoli by immunoelectron microscopy. Biol Cell 63: 27–34

    Google Scholar 

  • Thiry M, Ploton D, Ménager M, Goessens G (1993) Ultrastructural distribution of DNA within the nucleolus of various animal cell lines or tissues revealed by terminal deoxynucleotidyl transferase. Cell Tissue Res 271: 33–45

    Google Scholar 

  • Tyc K, Steitz JA (1989) U3, U8 and U13 comprise a new class of mammalian snRNPs localized in the cell nucleolus. EMBO J 8: 3113–3119

    Google Scholar 

  • Visa N, Puvion-Dutilleul F, Bachellerie JP, Puvion E (1993a) Intranuclear distribution of U1 and U2 snRNAs visualized by high resolution in situ hybridization: Revelation of a novel compartment containing U1 but not U2 snRNA in Hela cells. Eur J Cell Biol 60: 308–321

    Google Scholar 

  • Visa N, Puvion-Dutilleul F, Harper F, Bachellerie JP, Puvion E (1993b) Intranuclear distribution of Poly(A) RNA determined by electron microscopic in situ hybridization. Exp Cell Res 208: 19–34

    Google Scholar 

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

    Google Scholar 

  • Weibel E (1969) Stereological principles for morphometry in electron microscopic cytology. Int Rev Cytol 26: 235–302

    Google Scholar 

  • Williams L, Charest P, Fitzgerald G, Lafontaine JG (1985) A comparison of nuclease-gold over the nucleolus and nuclear bodies of Pisum sativum root tip cells. Biol Cell 54: 65–72

    Google Scholar 

  • Williams M (1977) Quantitative methods in biology. In: Glauert A (ed) Practical methods in electron microscopy 6. Elsevier, North-Holland American, pp 1–216

    Google Scholar 

  • Zamore P, Green MR (1991) Biochemical characterization of U2-snRNP auxiliary factor: An essential pre-mRNA splicing factor with a novel intranuclear distribution. EMBO J 10: 207–214

    Google Scholar 

  • Zareba-Kowalska A (1989) Cytochemical observations of the coiled bodies in neurons of rat sympathetic ganglia Histochemistry 91: 251–256

    Google Scholar 

Download references

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  1. Laboratory of Cellular and Tissular Biology, University of Liège, rue de Pitteurs, 20, B-4020, Liège, Belgium

    Marc Thiry

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Thiry, M. Cytochemical and immunocytochemical study of coiled bodies in different cultured cell lines. Chromosoma 103, 268–276 (1994). https://doi.org/10.1007/BF00352251

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  • DOI: https://doi.org/10.1007/BF00352251

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