Abstract
The shape of mitotic prophase chromosomes has been studied in root tip nuclei by confocal microscopy and 3D-image analysis. Crepis capillaris chromosome no. 1 was used as a test object. Chromosome conformation was studied in early, mid- and in late prophase. In mid- and late prophase, individual chromosomes could be distinguished on the basis of their length. Early prophase chromosomes could not be distinguished as individuals. The central axes of prophase chromosomes were traced with an automated computer procedure and then represented as a string of 3D coordinates. This representation facilitated measurement along the chromosome axis of shape parameters such as curvature (amount of bending), torsion (helical winding) and torsion sign (helical handedness). Stretches of early prophase chromosomes showed full helical turns, which could be left- or right-handed. In the later prophase stages curvature and torsion were statistically analysed. Our data on 40 mid-prophase chromosomes no. 1 show that they are still highly curved, but full helical turns were no longer found. Instead, an overall meandering pattern was observed. In late prophase, one central loop persisted, flanked by two preferential regions of high curvature.
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References
Agard DA, Sedat JW (1983) Three-dimensional architecture of a polytene nucleus. Nature 302: 676–681.
Bednar J, Horowitz RA, Grigoryev SA et al. (1998) Nucleosomes, linker DNA, and linker histone form a unique structural motif that directs the higher-order folding and compaction of chromatin. Proc Natl Acad Sci USA 95: 14173–14178.
Belmont AS, Braunfeld MB, Sedat JW, Agard DA (1989) Largescale chromatin structural domains within mitotic and interphase chromosomes in vivo and in vitro. Chromosoma 98: 129–143.
Boy de la Tour E, Laemmli UK (1988) The metaphase scaffold is helically folded: sister chromatids have predominantly opposite helical handedness. Cell 55: 937–944.
Brakenhoff GJ, van der Voort HTM, Oud JL (1990) Three-dimensional image representation in confocal microscopy. In: Wilson T ed. Confocal Microscopy. London: Academic Press, pp 185–197.
DuPraw EJ (1966) Evidence for a folded fibre organization in human chromosomes. Nature 209: 577–581.
Earnshaw WC (1988) Mitotic chromosome structure. BioEssays 9: 147–150.
Hiraoka Y, Minden JS, Swedlow JR, Sedat JW, Agard DA (1989) Focal points for condensation and decondensation revealed by three dimensional in vivo time-lapse microscopy. Nature 342: 293–296.
Hochstrasser M, Sedat JW (1987) Three dimensional organization of Drosophila melanogaster interphase nuclei. I. Tissue-specific aspects of polytene nuclear architecture. J Cell Biol 104: 1455–1470.
Houtsmuller AB, Smeulders AWM, van der Voort HTM, Oud JL, Nanninga N (1993) The homing cursor; a tool for three dimensional chromosome analysis. Cytometry 14: 501–509.
Kolmogorov A (1933) Sulla determinazione empirica di una legge di distribuzione. G Ist Ital Attuari 4: 83–91.
Luger K, Mäder AW, Richmond RK, Sargent DF, Richmond TJ (1997) Crystal structure of the nucleosome core particle at 2.8 A resolution. Nature 389: 251–260.
Manuelidis L (1990) A view of interphase chromosomes. Science 250: 1533–1540.
Manuelidis L, Chen TL (1990) A unified model of eukaryotic chromosomes. Cytometry 11: 8–25.
Marshall WF, Dernburg AB, Harmon B, Agard DA, Sedat JW (1996) Specific interactions of chromatin with the nuclear envelope: positional determination within the nucleus in Drosophila melanogaster. Mol Biol Cell 7: 825–842.
Montijn MB, Houtsmuller AB, ten Hoopen R, Oud JL, Nanninga N (1999) The 5S rRNA gene clusters have a defined orientation toward the nucleolus in Petunia hybrida and Crepis capillaris. Chromosome Res 7: 387–399.
Oud JL, Mans A, Brakenhoff GJ, van der Voort HTM, van Spronsen EA, Nanninga N (1989) Three-dimensional chromosome arrangement of Crepis capillaris in mitotic prophase and anaphase as studied by confocal scanning laser microscopy. J Cell Sci 92: 329–339.
Sedat JW, Manuelidis L (1977) A direct approach to the structure of eukaryotic chromosomes. Cold Spring Harbor Symp Quant Biol 42: 331–350.
Sumner AT (1990) Scanning electron microscopy of mammalian chromosomes from prophase to telophase. Chromosoma 100: 410–418.
van der Voort HTM, Brakenhoff GJ, Baarslag MW (1989) Three-dimensional visualization methods for confocal microscopy. J Microsc 153: 123–132.
Woltring HJ (1986) A Fortran package for generalized cross-validatory spline smoothing and differentiation. Adv Eng Software 8: 104–113.
Worring M, Pfluger P, Smeulders AWM, Houtsmuller AB (1994) Measurement of 3-D line shaped objects. Pat Rec Lett 15: 497–506.
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Houtsmuller, A.B., Oud, J.L., Montijn, M.B. et al. Chromosome No. 1 of Crepis Capillaris Shows Defined 3D-Shapes in Mitotic Prophase. Chromosome Res 8, 243–252 (2000). https://doi.org/10.1023/A:1009213332000
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DOI: https://doi.org/10.1023/A:1009213332000