Histochemistry and Cell Biology

, Volume 106, Issue 1, pp 59–78 | Cite as

Non-isotopic electron microscope in situ hybridization for studying the functional sub-compartmentalization of the cell nucleus

  • F. Puvion-Dutilleul
  • E. Puvion
Review

Abstract

Post-embedding electron microscope in situ hybridization using gold particles as label permits the clear identification of the cellular structures which contain the nucleic acid molecules under study. It has yielded information on the distribution of defined nucleic acid sequences of different origins—cellular or viral, DNA or RNA, single-or double-stranded molecules—which has revolutionized the study of the nucleus. Application of this powerful technique in combination with other refined techniques to studies on transcription and replication of cellular and viral genes has augmented our knowledge of the functional organization of the cell nucleus. One can now ask mechanistically meaningful questions concerning the successive steps of gene replication and expression not only under normal conditions of cell growth, but also when the cellular metabolism is altered by a drug treatment or a viral infection. This chapter aims (a) to present the established methods of post-embedding electron microscope in situ hybridization for localizing, precisely and specifically, a nucleic acid target in its normal environment and (b) to present some contributions of this technique to investigations of the functional compartmentalization of the cell nucleus and to elucidate the cell-virus relationships in infected cells.

Keywords

Gold Particle Herpes Simplex Virus Type Ribosomal Gene Granular Component Dense Fibrillar Component 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Azum-Gélade MC, Noaillac-Depeyre J, Caizergues-Ferrer M, Gas N (1994) Cell cycle redistribution of U3 snRNA and fibrillarin. Presence in the cytoplasmic nucleolus remnants and in the pre-nucleolar bodies at telophase J Cell Sci 107:463–475PubMedGoogle Scholar
  2. Bachmann L, Salpeter MM (1967) Absolute sensitivity of electron microscope radioautoradiography. J Cell Biol 33:299–305PubMedCrossRefGoogle Scholar
  3. Bassell GJ, Powers CM, Taneja KL, Singer RH (1994) Single mRNAs visualized by ultrastructural in situ hybridization are principally localized at actin filament intersections in fibroblasts. J Cell Biol 126:863–876PubMedCrossRefGoogle Scholar
  4. Besse S, Puvion-Dutilleul F (1994) Compartmentalization of cellular and viral DNAs in adenovirus type 5 ifnection as revealed by ultrastructural in situ hybridization. Chromosome Res 2:123–135PubMedCrossRefGoogle Scholar
  5. Besse S, Puvion-Dutilleul F (1995) Anchorage of adenoviral RNA to clusters of interchromatin granules. Gene Expression 5:79–92PubMedGoogle Scholar
  6. Besse S, Puvion-Dutilleul F (1996a) Intranuclear retention of ribosomal RNAs in response to herpes simples virus type 1 infection. J Cell Sci 109:119–129PubMedGoogle Scholar
  7. Besse S, Puvion-Dutilleul F (1996b) Distribution of ribosomal genes in nucleoli of herpes simplex virus type 1 infected cells. Eur J Cell Biol 70: in pressGoogle Scholar
  8. Besse S, Vigneron M, Pichard E, Puvion-Dutilleul F (1995) Synthesis and maturation of viral transcripts in herpes simplex virus type 1 infected HeLa cells. The role of interchromatin granules. Gene Expression 4:143–161PubMedGoogle Scholar
  9. Besse S, Diaz JJ, Pichard E, Kindbeiter K, Madjar JJ, Puvion-Dutilleul F (1996) In situ hybridization and immuno-electron microscope analysis of herpes simplex virus type 1 Us11 gene during transient expression. Chrosoma 104:434–444Google Scholar
  10. Bienz K, Egger D, Pfister T, Troxler M (1992) Structural and functional characterization of the poliovirus replication complex. J Virol 66:2740–2747PubMedGoogle Scholar
  11. Binder M (1992) In situ hybridization at the electron microscope level. In: Wilkinson DG (ed) In situ hybrydization: a practical approach. IRL Press, Oxford, pp 105–120Google Scholar
  12. Binder M, Tourmente S, Roth J, Renaud M, Gehring WJ (1986) In situ Lowicryl K4M-embedded tissue using biotinylated probes and protein A-gold complexes. J Cell Biol 102:1646–1653PubMedCrossRefGoogle Scholar
  13. Bond VC, Wold B, (1993) Nucleolar localization of myc transcripts. Mol Cell Biol 13:3221–3230PubMedGoogle Scholar
  14. Bowman LH, Rabin B, Schlessinger D (1981) Multiple ribosomal RNA cleavage pathways in mammalian cells. Nucleic Acids Res 9:4951–4966PubMedGoogle Scholar
  15. Brangeon J, Sossountzov L (1993) Electron microscope in situ hybridization to RNA or DNA in plant cells. In: Morel G (ed) Hybridization techniques for electron microscopy. CRC Press, Boca Raton, pp 301–348Google Scholar
  16. Cenacchi G, Musiani M, Gentilomi G, Righi S, Zerbini M, Chandler JG, Scala C, La Placa M, Martinelli GN (1993) In situ hybridization at the ultrastructural level: localization of cytomegalovirus DNA using digoxigenin labelled probes. J Submicrosc Cytol Pathol 25:341–345PubMedGoogle Scholar
  17. Chesselet MF (1990) In situ hybridization histochemistry. CRC Press, Boca Raton, pp 211Google Scholar
  18. Croissant O, Duguet C, Jeanteur P, Orth G (1972) Application de la technique d'hybridization moléculaire in situ à la mise en évidence au microscope électronique, de la réplication végétative de l'ADN viral dans les papillomes provoqués par le virus de Shope chez le lapin cottontail. C R Acad Sci Paris 274:614–617Google Scholar
  19. Dadoune JP, Siffroi JP, Alfonsi MF (1994) Ultrastructural localization or rDNA and rRNA by in situ hybridization in the nucleolus of human spermatids. Cell Tissue Res 278:611–616PubMedGoogle Scholar
  20. Dakashinamurti K, Mistry SP (1963) Tissue and intracellular distribution of biotin-C14OOH in rats and chicks. J Biol Chem 238:294–296Google Scholar
  21. De Furtos R, Kimura K, Peterson KR (1989) In situ hybridization ofDrosophila polytene chromosomes with digoxigenin-dUTP-labeled probes. Trens Genet 5:366–368CrossRefGoogle Scholar
  22. Dvorkin N, Clark MW, Hamkalo BA (1991) Ultrastructural localization of nucleic acid sequences inSaccharomyces cerevisiae nucleoli. Chromosoma 100:519–523PubMedCrossRefGoogle Scholar
  23. Epstein P, Reddy R, Busch H (1984) Multiple states of U3 RNA in Novikoff hepatoma nucleoli. Biochemistry 23:5421–5425PubMedCrossRefGoogle Scholar
  24. Escaig-Haye F, Grigoriev V, Fournier JG (1989) Détection ultrastructurale d'ARN ribosomal par hybridation in situ à l'aide d'une sonde biotinylée sur coupes ultrafines de cellule animale en culture. CR Acad Sci Paris 309:429–434Google Scholar
  25. Escaig-Haye F, Grigoriev V, Peranzi G, Lestienne P, Fournier JG (1991) Analysis of human mitochondrial transcripts using electron microscope in situ hybridization. J Cell Sci 100:851–862PubMedGoogle Scholar
  26. Escaig-Haye F, Grigoriev V, Sharova I, Rudneva V, Buckrinskaya A, Fournier JG (1992) Ultrastructural localization of HIV-1 RNA and core proteins. Simultaneous visualization using double immunogold labeling after in situ hybridization and immunocytochemistry. J Submicrosc Cytol Pathol 24:437–443PubMedGoogle Scholar
  27. Fakan S (1978) High resolution autoradiography studies on chromatin functions. In: Busch H (ed) The cell nucleus, vol 5. Academic Press, New York, pp 3–54Google Scholar
  28. Fakan S, Fakan J (1987) Autoradiography of spread molecular complexes. In: Sommerville J, Scheer U (eds) Electron microscopy in molecular biology: a practical approach. IRC Press, Oxford, pp 201–214Google Scholar
  29. Fournier JG, Escaig-Haye F (1993) Electron microscopy of rRNA in situ hybridization on Lowicryl sections. In: Morel G (ed) Hybridization techniques for electron microscopy. CRC Press, Boca Raton, pp 243–268Google Scholar
  30. Fournier MJ, Maxwell ES (1993) The nucleolar snRNAs: catching up with the spliceosomal snRNAs. Trends Biol Sci 18:131–135CrossRefGoogle Scholar
  31. Frappart L, Le Guellec D (1993) Ulstrastructural in situ hybridization of mRNA on Lowicryl sections using quick-freezing, freeze-substitution and low temperature embedding. In: Morel (ed) Hybridization techniques for electron microscopy. CRC Press, Boca Raton, pp 221–242Google Scholar
  32. Freund JN, Jost B, Duluc I, Morel G (1995) Ultrastructural study of intestinal lactase gene expression. Biol Cell 83: 211–218PubMedCrossRefGoogle Scholar
  33. Geuskens M, May E (1974) Ultrastructural localization of SV40 viral DNA in cells, during lytic infection, by in situ molecular hybridization. Exp Cell Res 87: 175–185PubMedCrossRefGoogle Scholar
  34. Hagari Y, Shibata M, Mihara M, Shimao S, Kurimura T (1993) Detection of human papillomavirus type 2a DNA in verrucae vulgares by electron microscopic in situ hybridization. Arch Dermatol Res 285: 255–260PubMedCrossRefGoogle Scholar
  35. Hamkalo NA, Narayanswami S, Lundgren K (1989) Localization of nucleic acid sequences by EM in situ hybridization using gold label. Am J Anat 185: 197–204PubMedCrossRefGoogle Scholar
  36. Hozak P, Schöfer C, Sylvester J, Wachtler F (1993) A study of nucleolar DNA: isolation of DNA from fibrillar components and ultrastructural localization of different DNA probes. J Cell Sci 104: 1199–1205PubMedGoogle Scholar
  37. Hutchison N, Langer-Safer P, Ward D, Hamkalo B (1982) In situ hybridization at the electron microscope level: hybrid detection by autoradiography and colloidal gold. J Cell Biol 95: 609–618PubMedCrossRefGoogle Scholar
  38. Jacob J, Tood K, Birnstiel ML, Bird A (1971) Molecular hybridization of3H labelled ribosomal RNA with DNA in ultrathin sections prepared for electron microscopy. Biochim Biophys Acta 228: 761–776PubMedGoogle Scholar
  39. Jiao R, Yu W, Ding M, Zhai Z (1992) Localization of adenovirus DNA by in situ hybridization electron microscopy. Microsc Res Tech 21: 23–31PubMedCrossRefGoogle Scholar
  40. Jimenez-Garcia LF, Segura-Valdez ML, Ochs RL, Echeverria OM, Vasquez-Nin GH, Busch H (1993) Electron microscopic localization of ribosomal DNA in rat liver nucleoli by non-isotopic in situ hybridization. Exp Cell Res 207: 220–225PubMedCrossRefGoogle Scholar
  41. John HA, Birnstiel ML, Jones KW (1969) RNA-DNA hybrids at a cytological level. Nature 223: 582–587PubMedCrossRefGoogle Scholar
  42. Kalland KH, Langhoff E, Bos HJ, Gottlinger H, Haselting WA (1991) REX-dependent nucleolar accumulation of HTLV-1 mRNA. New Biol 3: 389–397PubMedGoogle Scholar
  43. Kadowaki T, Schneiter R, Hitomi M, Tartakoff AM (1995) Mutations in nucleolar proteins lead to nucleolar accumulation of polyA+ RNA inSaccharomyces cerevisiae. Mol Biol Cell 6: 1103–1110PubMedGoogle Scholar
  44. Langer PR, Waldrop AA, Ward DC (1981) Enzymatic synthesis of biotin labeled polynucleotides: novel nucleic acid affinity probes. Proc Natl Acad Sci USA 78: 6633–6637PubMedCrossRefGoogle Scholar
  45. Langer-Safer PR, Levine M, Ward DC (1982) Immunological methods for mapping genes onDrosophila polytene chromosomes. Proc Natl Acad Sci USA 79: 4381–4385PubMedCrossRefGoogle Scholar
  46. Le Guellec D, Frappart L, Desprez PY (1990) Ultrastructural localization of mRNA encoding for the EGF receptor in human breast cell line BT20 by in situ hybridization. J Histochem Cytochem 39: 1–6Google Scholar
  47. Leitch AR, Mosgöller W, Schwarzacher T, Bennett MD, Heslop-Harrison JS (1990) Genomic in situ hybridization to sectioned nuclei shows chromosome domains in grass hybrids. J Cell Sci 95: 335–341PubMedGoogle Scholar
  48. Lin NS, Chen CC, Hsu YH (1993) Post-embedding in situ hybridization for localization of viral nucleic acid in ultra-thin sections. J Histochem Cytochem 41: 1513–1519PubMedGoogle Scholar
  49. Mandry P, Murray AB, Rieke L, Becke H, Höfler H (1993) Post-embedding ultrastructural in situ hybridization on ultrathin cryosections and LR White resin sections. Ultrastruct Pathol 17: 185–194PubMedGoogle Scholar
  50. McFadden GI (1989) In situ hybridization in plants: from microscopic to ultrastructural resolution. Cell Biol Int Rep 13: 3–21CrossRefGoogle Scholar
  51. McFadden GI, Bonig I, Cornish EC, Clarke AE (1988) A simple fixation and embedding method for use in hybridization histochemistry on plant tissues. Histochem J 20: 575–586PubMedCrossRefGoogle Scholar
  52. McFadden G, Bönig I, Clarke A (1990) Double label in situ hybridization for electron microscopy. Trans R Microsc Soc 1: 683–688Google Scholar
  53. Medina FJ, Cerdido A, Fernandez-Gomez ME (1995) Components of the nucleolar processing complex (pre-rRNA, fibrillarin, and nucleolin) colocalize during mitosis and are incorporated to daughter cell nucleoli. Exp Cell Res 221: 111–125PubMedCrossRefGoogle Scholar
  54. Morel G (1991) Hybridation in situ sur coupes ultrafines de tissus congelés. Sonde radioactive, sonde non radioactive, double marquage. In: Morel G (ed) Microscopie électronique: cryométhodes, immunocytologie, autoradiographie, hybridation in situ. Editions INSERM, Paris, pp 519–534Google Scholar
  55. Morel G (1993) In situ hybridization on ultrathin frozen sections. In: Morel G (ed) Hybridization techniques for electron microscopy. CRC Press, Boca Raton, pp 163–220Google Scholar
  56. Morel G, Dubois PM, Gossard F (1986) Détection ultrastructurale des ARN messagers codant pour l'hormone de croissance dans l'antehypophyse du rat par hybridation in situ. C R Acad Sci Paris 302: 479–484PubMedGoogle Scholar
  57. Morey AL, (1995) Non-isotopic in situ hybridization at the ultrastructural level. J Pathol 176: 113–121PubMedCrossRefGoogle Scholar
  58. Morey AL, Ferguson DJ, Leslie KO, Taatjes DL, Fleming KA (1993) Intracellular localization of parvovirus B19 nucleic acid at the ultrastructural level by in situ hybridization with digoxigenin-labelled probes. Histochem J 25: 421–429PubMedCrossRefGoogle Scholar
  59. Morey AL, Ferguson DJ, Fleming KA (1995) Combined immunohistochemistry and non-isotopic in situ hybridization for ultrastructural analysis of human parvovirus B19 infection. Histochem J 27: 46–53PubMedCrossRefGoogle Scholar
  60. Mosgöller W, Schöfer C, Derenzini M, Steiner M, Maier U, Wachtler F (1993) Distribution of DNA in human Sertoli cell nucleoli. J Histochem Cytochem 41: 1487–1493PubMedGoogle Scholar
  61. Multhaupt HAB, Rafferty PA, Warhol MJ (1992) Ultrastructural localization of human papilloma virus by nonradioactive in situ hybridization on tissue of human cervical intraepithelial neoplasia. Lab Invest 67: 512–518PubMedGoogle Scholar
  62. Narayanswami S, Hamkalo BA (1987) Hybridization to chromatin and whole chromosome mounts. In: Sommerville J, Scheer U (eds) Electron microscopy in molecular biology: a practical approach. IRL Press, Oxford, pp 215–232Google Scholar
  63. Niedobitek G, Finn T, Herbst H, Stein H (1989) Detection of viral genomes in the liver by in situ hybridization using35S-bromodeoxyuridine and biotin-labeled probes. Am J Pathol 134: 633–639PubMedGoogle Scholar
  64. Olmedilla A, Testillano PS, Vicente O, Delseny M, Risueno MC (1993) Ultrastructural rRNA localization in plant nucleoli. RNA/RNA in situ hybridization, autoradiograph and cytochemistry. J Cell Sci 106: 1333–1346PubMedGoogle Scholar
  65. O'Reilly MM, French SL, Sikes ML, Miller OL Jr (1994) Ultrastructural in situ hybridization to nascent transcripts of highly transcribed rRNA genes in chromatin spreads. Chromosoma 103: 122–128PubMedGoogle Scholar
  66. Pardue ML, Gall JG (1969) Molecular hybridization of radioactive DNA to the DNA of cytological preparations. Proc Natl Acad Sci USA 64: 600–604PubMedCrossRefGoogle Scholar
  67. Pautrat G, Morel G, Dihl F, Allasia C, Rey F, Spire J, Kourilsky F Chermann JG (1988) Distribution polaire des particules virales à la surface des cellules d'une lignéee lymphocytaire infectées in vitro par le virus HIV: détection intracellulaire du génome viral. Retrovirus 47: 511–517Google Scholar
  68. Pierron G, Puvion-Dutilleul F (1993) Mitotic segregation of the nucleolar ribosomal RNA inPhysarum polycephalum. Exp Cell Res 208: 509–517PubMedCrossRefGoogle Scholar
  69. Pollock JA, Ellisman MH, Benzer S (1990) Subcellular localization of transcripts inDrosophila photoreceptor neurons: chaoptic mutants have an aberrant distribution. Genes Dev 4: 806–821PubMedGoogle Scholar
  70. Puvion E, Moyne G (1981) In situ RNA structures. In: Busch H (ed) The cell nucleus, vol 8. Academic Press, New York, pp 59–115Google Scholar
  71. Puvion-Dutilleul F (1993a) Procedures of in situ nucleic acid hybridization to detect viral DNA and RNA in cells by electron microscopy. In: Morel G (ed) Hybridization techniques for electron microscopy. CRC Press, Boca Raton, pp 270–299Google Scholar
  72. Puvion-Dutilleul F (1993b) Protocol of electron microscope in situ nucleic acid hybridization for the exclusive detection of double-stranded DNA sequences in cells containing large amounts of homologous single-stranded DNA and RNA sequences. Application to adenovirus type 5 infected HeLa cells. Microsc Res Tech 25: 2–11PubMedCrossRefGoogle Scholar
  73. Puvion-Dutilleul F (1995) Visualization and identification of viral nucleic acids. In: Morel G (ed) Visualization of nucleic acids. CRC Press, Boca Raton, pp 229–321Google Scholar
  74. Puvion-Dutilleul F, Besse S (1994) Induction of complete segregation of cellular DNA and non-encapsidated viral genomes in herpes simplex virus type 1 infected HeLa cells as revealed by in situ hybridization. Chromosoma 103: 104–110PubMedGoogle Scholar
  75. Puvion-Dutilleul F, Pichard E (1992) Segregation of viral double-stranded and single-stranded DNA molecules in nuclei of adenovirus infected cells as revealed by electron microscope in situ hybridization. Biol Cell 76: 139–150PubMedCrossRefGoogle Scholar
  76. Puvion-Dutilleul F, Pierron G (1992) Localization by high resolution in situ hybridization of the ribosomal minichromosomes during the nucleolar cycle ofPhysarum polycephalum Exp Cell Res 203: 354–364PubMedCrossRefGoogle Scholar
  77. Puvion-Dutilleul F, Puvion E (1989) Ultrastructural localization of viral DNA in thin sections of herpes simplex virus type 1 infected cells by in situ hybridization. Eur J Cell Biol 49: 99–109PubMedGoogle Scholar
  78. Puvion-Dutilleul E, Puvion E (1990a) Replicating single-stranded adenovirus type 5 DNA molecules accumulate within well-delimited intranuclear areas of lytically infected HeLa cells. Eur J Cell Biol 52: 379–388PubMedGoogle Scholar
  79. Puvion-Dutilleul F, Puvion E (1990b) Analysis by in situ hybridization and autoradiography of sites of replication and storage of single- and double-stranded adenovirus type 5 DNA in lytically infected HeLa cells. J Struct Biol 103: 280–289PubMedCrossRefGoogle Scholar
  80. Puvion-Dutilleul F, Puvion E (1991a) Ultrastructural localization of defined sequences of viral RNA and DNA by in situ hybridization of biotinylated DNA probes on sections of herpes simplex virus type 1 infected cells. J Electron Microsc Tech 18: 336–353PubMedCrossRefGoogle Scholar
  81. Puvion-Dutilleul F, Puvion E (1991b) Sites of transcription of adenovirus type 5 genomes in relation to early viral DNA replication in infected HeLa cells. A high resolution in situ hybridization and autoradiographical study. Biol Cell 71: 135–147PubMedCrossRefGoogle Scholar
  82. Puvion-Dutilleul F, Puvion E (1995) Immunocytochemistry, autoradiography, in situ hybridization, selective stains: complementary tools for ultrastructural study of structure-function relationships in the nucleus. Applications to adenovirus-infected cells. Microsc Res Tech 31: 22–43PubMedCrossRefGoogle Scholar
  83. Puvion-Dutilleul F, Pichard E, Laithier M, Puvion E (1989) Cytochemical study of the localization and organization of parental herpes simplex virus type 1 DNA during initial infection of the cell. Eur J Cell Biol 50: 187–200PubMedGoogle Scholar
  84. Puvion-Dutilleul F, Bachellerie JP, Puvion E (1991a) Nucleolar organization of HeLa cells as studied by in situ hybridization. Chromosoma 100: 395–409PubMedCrossRefGoogle Scholar
  85. Puvion-Dutilleul F, Mazan S, Nicoloso M, Christensen ME, Bachellerie JP (1991b) Localization of U3 RNA molecules in nucleoli of HeLa and mouse 3T3 cells by high resolution in situ hybridization. Eur J Cell Biol 56: 178–186PubMedGoogle Scholar
  86. Puvion-Dutilleul F, Mazan S, Nicoloso N, Pichard E, Bachellerie JP, Puvion E (1992a) Alteration of nucleolar ultrastructure and ribosome biogenesis by actinomycin D. Implications for U3 snRNP function. Eur J Cell Biol 58:149–162PubMedGoogle Scholar
  87. Puvion-Dutilleul F, Roussev R, Puvion E, (1992b) Distribution of viral RNA molecules during the adenovirus type 5 infectious cycle in HeLa cells. J Struct Biol 108:209–220PubMedCrossRefGoogle Scholar
  88. Puvion-Dutilleul F, Bachellerie JP, Visa N, Puvion E (1994) Rearrangements of intranuclear structures involved in RNA processing in response to adenovirus infection. J Cell Sci 107:1457–1468PubMedGoogle Scholar
  89. Puvion-Dutilleul F, Besse S, Chan EK, Tan EM, Puvion E (1995) p-80 Coilin: a component of coiled body and interchromatin in granule-associated zones. J Cell Sci 108:1143–1153PubMedGoogle Scholar
  90. Raska I, Dundr M, Koberna K, Melcak I, Risueno CM, Torok I (1995) Does the synthesis of ribosomal RNA take place within nucleolar fibrillar centers or dense fibrillar components? A critical appraisal. J Struct Biol 114:1–22PubMedCrossRefGoogle Scholar
  91. Salpeter MM, Bachmann L, Salpeter EE (1969) Resolution in electron microscope autoradiography. J Cell Biol 41:1–20PubMedCrossRefGoogle Scholar
  92. Stahl A, Wachtler F, Hartung M, Devictor M, Schöfer C, Mösgoller W, Lanversin A de, Fouet C, Schwarzacher HG (1991) Nucleoli, nucleolar chromosomes and ribosomal genes in the human spermatocyte. Chromosoma 101:231–244PubMedCrossRefGoogle Scholar
  93. Steinert G, Thomas C, Brachet J (1976) Localization by in situ hybridization of amplified ribosomal DNA duringXenopus laevis maturation (a light and electron microscopic study). Proc Natl Acad Sci USA 73:833–836PubMedCrossRefGoogle Scholar
  94. Tani T, Berby RJ, Hiraoka Y, Spector DL (1995) Nucleolar accumulation of poly(A)+ RNA in heat-shocked yeast cells: implication of nucleolar involvement in mRNA transport. Mol Biol Cell 6:1515–1534PubMedGoogle Scholar
  95. Testillano PS, Gonzalez-Melendi P, Ahmadian P, Fadon B, Risueno MC (1995) The immunolocalization of nuclear antigens during the pollen developmental program and the induction of pollen embryogenesis. Exp Cell Res 221:41–54PubMedCrossRefGoogle Scholar
  96. Thiry M (1992) New data concerning the functional organization of the mammalian cell nucleolus: detection of RNA and rRNA by in situ molecular immunocytochemistry. Nucleic Acids Res 20:6195–6200PubMedGoogle Scholar
  97. Thiry M, 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 50:235–243PubMedGoogle Scholar
  98. Thiry M, Thiry-Blaise L (1991) Locating transcribed and nontranscribed rDNA spacer sequences within the nucleolus by in situ hybridization and immunoelectron microscopy. Nucleic Acids Res 19:11–15PubMedGoogle Scholar
  99. Trembleau A, Fevre-Montange M, Calas A (1988) Localisation ultrastructurale d'ARNm codant pour l'ocytocine par hybridation in situ. Etude par radioautographie à haute résolution à l'aide d'une sonde oligonucléotidique tritiée. C R Acad Sci Paris 307:869–874PubMedGoogle Scholar
  100. Trembleau A, Calas A, Fevre-Montange M (1990) Ultrastructural localization of oxytocin mRNA in the rat hypothalamus by in situ hybridization using synthetic oligonucleotide. Mol Brain Res 8:37–45PubMedCrossRefGoogle Scholar
  101. Troxler M, Pasamontes L, Egger D, Bienz K (1990) In situ hybridization for light and electron microscopy: a comparison of methods for the localization of viral RNA using biotinylated DNA and RNA probes. J Virol Methods 30:1–14PubMedCrossRefGoogle Scholar
  102. Troxler M, Egger D, Pfister T, Bienz K (1992) Intracellular localization of poliovirus RNA by in situ hybridization at the ultrastructural level using single-stranded riboprobes. Virology 191:687–697PubMedCrossRefGoogle Scholar
  103. Visa N, Puvion-Dutilleul F, Bachellerie JP, Puvion E (1993a) Intranuclear distribution of U1 and U2 snRNAs as 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: 308–321PubMedGoogle Scholar
  104. Visa N, Puvion-Dutilleul F, Harper F, Bachellerie JP, Puvion E (1993b) Intranuclear distribution of poly(A) RNA determined by electron microscope in situ hybridization. Exp Cell Res 208: 19–34PubMedCrossRefGoogle Scholar
  105. Wachtler F, Schöfer C, Mosgöller W, Weipoltshammer K, Schwarzacher HG, Guichaoua M, Hartung M, Stahl A, Bergé-Lefranc JL, Gonzalez I, Sylvester J (1992) Human ribosomal RNA gene repeats are localized in the dense fibrillar component of nucleoli—light and electron microscopic in situ hybridization in human Sertoli cells. Exp Cell Res 198: 135–143PubMedCrossRefGoogle Scholar
  106. Wenderoth MP, Eisenberg BR (1991) Ultrastructural distribution of myosin heavy chain mRNA in cardiac tissue: a comparison of frozen and LR White embedment. J Histochem Cytochem 39: 1025PubMedGoogle Scholar
  107. Wolber RA, Beals TF, Lloyd RV, Maassab HF (1988) Ultrastructural localization of viral nucleic acid by in situ hybridization. Lab Invest 59: 144–151PubMedGoogle Scholar
  108. Wolber RA, Beals T, Maassab HF (1989) Ultrastructural localization of herpes simplex virus RNA by in situ hybridization. J Histochem Cytochem 37: 97–104PubMedGoogle Scholar
  109. Yao CH, Kitazawa S, Fujimori T, Maeda S (1993) In situ hybridization at the electron microscope level using a bromodeoxyuridine labeled DNA probe. Biotech Histochem 68: 169–174PubMedGoogle Scholar
  110. Yi J, Michel O, Sassy-Prigent C, Chevalier J (1995) Electron microscopic location of mRNA in the rat kidney: improved post-embedding in situ hybridization. J Histochem Cytochem 43:801–809PubMedGoogle Scholar
  111. Yun K, Sherwood MJ (1992) In situ hybridization at light and electron microscopic levels: identification of human papillo-mavirus nucleic acids. Pathology 24: 91–98PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • F. Puvion-Dutilleul
    • 1
  • E. Puvion
    • 1
  1. 1.Laboratoire Organisation fonctionelle du Novau de l'UPR 9044 CNRSVillejuif CedexFrance

Personalised recommendations