Skip to main content
SpringerLink
Log in

Three-dimensional analysis of the arrangement of compact chromatin in the nucleus of G0 rat lymphocytes

  • Original Paper
  • Published:
Histochemistry and Cell Biology Aims and scope Submit manuscript

Abstract

The arrangement of compact chromatin of G0 lymphocytes was studied in three-dimensional reconstructions of the ensemble of the chromatin and of individual compact chromatin bodies. Rat spleen was serially cut and sections were contrasted with procedures preferential for DNA. Electron microscopy images were digitized, processed, and displayed using a commercial soft-ware package, complemented by a system for three-dimensional reconstruction and analysis developed by us on an IBM-compatible microcomputer provided with an image acquisition board. The reconstructions showed a continuous layer of compact chromatin in contact with the nuclear envelope that prevents the automatic recognition of individual chromatin clumps. The ensemble of the arrangement of compact chromatin was found to be very similar in different lymphocytes. After morphological filtering procedures, the initial mass was divided into individual bodies of compact chromatin, which were tagged. Most of these bodies contact the nuclear envelope. The number of bodies as well as the number of contacts with the envelope are similar and correspond to a haploid number of chromosomes. The largest body is always the one containing nucleolus-associated chromatin. When the cell has two nucleoli, the nucleolus-associated chromatin bodies contact the envelope in diametrically opposed areas. This feature was also described in rat liver cells. It is concluded that: (a) the individualized compact chromatin bodies do not correspond to an entire chromosome or to a pair of chromosomes; (b) the arrangement of compact chromatin is not identical in each G0 lymphocyte, but there are patterns that are repeated with limited changes; and (c) there are common features that appear in different cell types of individuals of the same species.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Appels R, Driscoll C, Peacock WJ (1978) Heterochromatin and highly repeated DNA sequences in rye (Secale cereale). Chromosoma 70:67–89

    Google Scholar 

  • Artzy E, Frieder G, Herman G (1981) The theory, design, implementation, and evaluation of a three-dimensional surface detection algorithm. Comput Vision Graphics Image Processing 15:1–24

    Google Scholar 

  • Avivi L, Feldman M (1980) Arrangement of chromosomes in the interphase nucleus of plants. Hum Genet 55:281–295

    PubMed  Google Scholar 

  • Baker H, Lindsey J, Weisbroth S (1979) The laboratory rat. Academic Press, New York

    Google Scholar 

  • Bennett MD (1982) Nucleotypic basis of the spatial ordering of chromosomes in eukaryotes and implications of the order for genome evolution and phenotypic variation. In: Dover GA, Flavell RB (eds) Genome evolution, Academic Press London, pp 239–261

    Google Scholar 

  • Bennett MD (1984) Nuclear architecture and its manipulation. In: Gustafson JP (ed) Gene manipulation in plant improvement. Plenum, New York, pp 469–501

    Google Scholar 

  • Beucher S, Lantuejoul C (1979) Use of watersheds in contour detection. In: Proceedings of the International Workshop on Image Processing: real-time edge and motion detection/estimation. Sponsored by: Centre Commun des Études en Télévision et Télécommunications. Institut National des Sciences Appliquées et Institut de Recherche en Informatique, et Systèmes Aleatoires. Rennes, France, 1–12 February

  • Bochkov NP, Lopukhin YM, Kuleshov NP, Kovalchuk LV (1974) Cytogenetic study of patients with ataxia-teleangiectasia. Humangenetik 24:115–128

    PubMed  Google Scholar 

  • Bourgeois CA, Laquerriere F, Hemon D, Hubert J, Bouteille M (1985) New data on in situ position of the inactive X chromosonme in the interphase nucleus of human fibroblasts. Hum Genet 69:122–129

    PubMed  Google Scholar 

  • Brenner S, Pepper D, Berns MW, Tan E, Brinkley BR (1981) Kinetochore structure, duplication, and distribution in mammalian cells: analysis by human autoantibodies from scleroderma patients. J Cell Biol 91:95–102

    PubMed  Google Scholar 

  • 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 Acad Sci D 276:3041–3044

    Google Scholar 

  • Cohen MM, Shaw MW (1964) Effects of mitomycin C on human chromosomes. J Cell Biol 23:386–395

    PubMed  Google Scholar 

  • Comings DE (1968) The rationale for an order arrangement of chromatin in interphase nucleus. Am J Genet 20:440–460

    Google Scholar 

  • Comings DE (1980) Arrangement of chromatin in the nucleus. Hum Genet 53:331–343

    Google Scholar 

  • Cremer T, Cremer C, Baumann H, Luedtke EK, Sperling K, Teuber V, Zorn C (1982) Rabl's model of the interphase chromosome arrangement tested in Chinese hamster cells by premature chromosome condensation and laser-UV microbeam experiments. Hum Genet 60:46–56

    PubMed  Google Scholar 

  • Cremer T, Tesin D, Hopman AH, Manuelidis L (1988) Rapid interphase and metaphase assessment of specific chromosomal changes in neuroectodermal tumor cells by in situ hybridization with chemically modified DNA probes. Exp Cell Res 176:199–220

    PubMed  Google Scholar 

  • DeBoni U, Mintz AH (1986) Curvilinear, three-dimensional motion of chromatin domains and nucleoli in neuronal interphase nuclei. Science 234:863–866

    PubMed  Google Scholar 

  • Esquivel C, Rovira P, Echeverría O, Vázquez Nin GH (1987) A simple staining method for chromatin in electron microscopy compatible with serial sectioning. Ultramicroscopy 21:103–110

    PubMed  Google Scholar 

  • Esquivel C, Vázquez Nin GH, Echeverría O (1989) Evidence of repetitive patterns of chromatin distribution in cell nuclei of rat liver. Acta Anat 136:94–98

    PubMed  Google Scholar 

  • Evans JK, Filion WG (1982) The distribution of chromatin in the interphase nucleus ofZebrina pendula. Can J Genet Cytol 24: 583–591

    Google Scholar 

  • Ferguson M, Ward DC (1992) Cell cycle dependent movement in pre-mitotic human T-lymphocyte nuclei. Chromosoma 101: 557–565

    PubMed  Google Scholar 

  • Fussell CP (1975) The position of interphase chromosomes and late replicating DNA in centromere and telomere regions ofAllium cepa L. Chromosoma 50:201–210

    Google Scholar 

  • Fussell CP (1977) Telomere associations in interphase nuclei ofAllium cepa demonstrated by C-banding. Exp Cell Res 110: 111–117

    PubMed  Google Scholar 

  • Ghosh S, Roy SCh (1977) Orientation of interphase chromosomes as detected by Giemsa C-bands. Chromosoma 61:49–55

    PubMed  Google Scholar 

  • Goldman IL, Zolotarev VM, Vasileva SF (1977) The space location of chromosomes in the somatic cell nuclei. IV. Homologous chromosomes in guinea pig and domestic pig cells. Tsitologiia 19:855–863

    PubMed  Google Scholar 

  • Gordon D, Udupa J (1989) Fast surface tracking in three-dimensional binary images. Comput Vision Graphics Image Processing 45:196–214

    Google Scholar 

  • Hadlaczky GY, Went M, Ringertz NR (1986) Direct evidence for non-random localization of mammalian chromosomes in the interphase nucleus. Exp Cell Res 167:1–15

    PubMed  Google Scholar 

  • Hager HD, Schoeder-Kurth TM, Vogel F (1982) Position of chromosomes in the human interphase nucleus. An analysis of nonhomologous chromatid translocations in lymphocyte cultures after Treminon treatment and from patients with Franconi's anemia and Bloom's syndrome. Hum Genet 61:342–356

    PubMed  Google Scholar 

  • Hilliker AJ, Appels R (1989) The arrangement of interphase chromosomes: structural and functional aspects. Exp Cell Res 185: 297–318

    Google Scholar 

  • Holowacz T, DeBoni U (1991) Arrangement of kinetochore proteins and satellite DNA in neuronal interphase nuclei changes induced by gamma-aminobutyric acid (GABA). Exp Cell Res 197:36–42

    PubMed  Google Scholar 

  • Hubert J, Bourgeois CA (1986) The nuclear skeleton and the spatial arrangement of chromosomes in the interphase nucleus of vertebrate somatic cells. Hum Genet 74:1–15

    PubMed  Google Scholar 

  • Kuroiwa T, Tanaka N (1971a) Fine structure of interphase nucleus. I. The morphological classification of nucleus in interphase ofCrepis capillaris. Cytologia 36:143–160

    Google Scholar 

  • Kuroiwa T, Tanaka N (1971b) Fine structure of interphase nucleus. IV. The behavior of late replicating chromatin during a late portion of S period as revealed by electron microscope radio-autography. J Cell Biol 49:939–942

    PubMed  Google Scholar 

  • Lafontaine JG, Lord A (1974) An ultrastructural and radioautographic study of the evolution of the interphase nucleus in plant meristemic cells (Allium porrum). J Cell Sci 14:263–287

    PubMed  Google Scholar 

  • Leger I, Guillaud M, Krief B, Brugal G (1994) Interactive computer-assisted analysis of chromosome I colocalization with nucleoli. Cytometry 16:313–323

    PubMed  Google Scholar 

  • Leitch AR, Mosgöller W, Scwarzacher T, Bennett MD (1990) Genomic in situ hybridization to sectioned nuclei shows chromosome domains in grass hybrids. J Cell Sci 95:335–341

    PubMed  Google Scholar 

  • Lyon MF (1972) X-chromosome inactivation and developmental patterns in mammals. Biol Rev 47:1–35

    PubMed  Google Scholar 

  • Manuelidis L (1984) Different CNS cell types display distinct and nonrandom arrangements of satellite DNA sequences. Proc Natl Acad Sci USA 81:3123–3127

    PubMed  Google Scholar 

  • Manuelidis L (1985a) Indications of centromere movement during interphase and differentiation. Ann NY Acad Sci 450:205–221

    PubMed  Google Scholar 

  • Manuelidis L (1985b) Individual interphase chromosome domains revealed by in situ hybridization. Hum Genet 71:288–293

    PubMed  Google Scholar 

  • Manuelidis L, Langer-Safer, PR, Ward DC (1982) High-resolution mapping of satellite DNA using biotin-labeled DNA probes. J Cell Biol 95:619–625

    PubMed  Google Scholar 

  • Mares V, Giordano PA, Pelliciari C, Scherini E, Lisá V, Bottone MG, Bottiroli G (1991) Changes in cell cycle and chromatin distribution in C6 glioma cells treated by dibutyryl cyclic AMP. Cell Prolif 24:569–577

    PubMed  Google Scholar 

  • Metz CW (1916) Chromosome studies on the Diptera. II. The paired association of chromosomes in the Diptera and its significance. J Exp Zool 21:213–279

    Google Scholar 

  • Moroi Y, Hartman AL, Nakane PK, Tan EM (1981) Distribution of kinetochore (centromere) antigen in mammalian cell nuclei. J Cell Biol 90:254–259

    PubMed  Google Scholar 

  • Murray AB, Davies HG (1979) Three-dimensional reconstruction of the chromatin bodies in the nuclei of mature erythrocytes from the newtTriturus cristatus: the number of nuclear envelope-attachment sites. J Cell Sci 35:59–66

    PubMed  Google Scholar 

  • Park PC, DeBoni U (1992) Spatial rearrangement and enhanced clustering of kinetochores in interphase nuclei of dorsal root ganglion neurons in vitro: association with nucleolar fusion. Exp Cell Res 203:222–229

    PubMed  Google Scholar 

  • Pomerat CM, Hendelman WJ, Raiborn CW, Massey JF (1967) Dynamic activities of nervous tissue in vitro. In: Hyden H (ed) The neuron Elsevier New York, pp 119–173

    Google Scholar 

  • Rabl C (1885) Über Zellteilung. Morphol Jahrb 10:214–330

    Google Scholar 

  • Schardin M, Cremer T, Hager HD, Lang M (1985) Specific staining of human chromosomes in Chinese hamster x man hybrid cell lines demonstrates interphase chromosome territories. Hum Genet 71:281–287

    PubMed  Google Scholar 

  • Stack S, Roy SCh (1977) Orientation of interphase chromosomes as detected by Giemsa C-bands. Chromosoma 61:49–55

    PubMed  Google Scholar 

  • Udupa J (1981) Determination of 3-D shape parameters from boundary information. Comput Vision Graphics Image Processing 17:52–59

    Google Scholar 

  • Udupa J (1990) Boundary and object labelling in three-dimensional images. Comput Vision Graphics Image Processing 5:355–369

    Google Scholar 

  • Vázquez Nin GH, Chávez B, Tomás-Martin C (1973) A preferential staining method for chromatin in electron microscopy. J Microsc 16:243–246

    Google Scholar 

  • Vázquez Nin GH, Echeverría OM, Molina E, Fragoso J (1978) Effects of ovariectomy and estradiol injection on nuclear structures of endometrial epithelial cells. Acta Anat 102:308–318

    PubMed  Google Scholar 

  • Vogel F, Schoeder TM (1974) Internal order of the interphase nucleus. Humangenetik 25:265–297

    PubMed  Google Scholar 

  • Testillano PS, Sánchezpina MA, Olmedilla A, Tandler CJ, Risueño MC (1991) A specific ultrastructural method to reveal DNA. The NAMA-Ur. J Histochem Cytochem 39:1427–1438

    PubMed  Google Scholar 

  • Wachtler F, Hopman AHN, Wiegant J, Schwarzacher HG (1986) On the position of nucleolus organizer regions (NORs) in interphase nuclei. Studies with a new, non-autoradiographic in situ hybridization method. Exp Cell Res 167:227–240

    PubMed  Google Scholar 

  • Weimer R, Haaf T, Krüger J, Poot M, Schmid M (1992) Characterization of centromere arrangements and test for random distribution in G0, G1, S, G2, G1, and early S′ phase in human lymphocytes. Hum Genet 88:673–682

    PubMed  Google Scholar 

  • Westergaard M (1964) Studies on the mechanism of crossing over. I. Theoretical considerations. C R Trav Lab Calrsberg 34:359–405

    Google Scholar 

  • Wettstein R, Graver A (1973) Film-supporting frames for mounting section grids. J Ultrastruct Res 43:436–447

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratory of Electron Microscopy, Department of Biology, Faculty of Sciences, UNAM, Apartado Postal 70-438, México, DF, Mexico

    Gabriel López-Velázquez, Ernestina Ubaldo, Olga Echeverría & Gerardo H. Vázquez Nin

  2. Laboratory of Image Processing, Center of Instruments, UNAM, Mexico

    Jorge Márquez & Gabriel Corkidi

Authors
  1. Gabriel López-Velázquez
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jorge Márquez
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ernestina Ubaldo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gabriel Corkidi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Olga Echeverría
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Gerardo H. Vázquez Nin
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

López-Velázquez, G., Márquez, J., Ubaldo, E. et al. Three-dimensional analysis of the arrangement of compact chromatin in the nucleus of G0 rat lymphocytes. Histochem Cell Biol 105, 153–161 (1996). https://doi.org/10.1007/BF01696155

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01696155

Keywords

Search

Navigation