Abstract
This report describes a fluorescence in situ hybridization approach to chromosome staining that facilitates detection of structural aberrations and allows discrimination between dicentric chromosomes and symmetrically translocated chromosomes. In this approach, selected whole chromosomes are stained in one color by hybridization with composite probes whose elements have DNA sequence homology along the length of the target chromosomes. In addition, all chromosomes are counterstained with a DNA specific dye so that structural aberrations between target and non-target chromosomes are clearly visible. Discrimination between dicentric chromosomes and symmetrical translocations is accomplished by hybridization with a second probe that is homologous to DNA sequences found in the centromeric region of all chromosomes. The centromeric marker is visualized in a different color, so that the number of centromeres per aberrant chromosome can be rapidly determined in the microscope by changing excitation and fluorescence filters.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Awa AA (1983) Chromosome damage in atomic bomb survivors and their offspring — Hiroshima and Nagasaki. In: Ishihara T, Sasaki MS (eds) Radiation induced chromosome damage in man. Liss, New York, pp 433–453
Bauchinger M (1984) Cytogenetic effects in human lymphocytes as a dosimetry system. In: Eisert WG, Mendelsohn ML (eds) Biological dosimetry. Springer, Berlin Heidelberg New York, pp 15–24
Buckton KE, Hamilton GE, Paton L, Langlands AO (1978) Chromosome aberrations in irradiated ankylosing spondylitis patients. In: Evans HJ, Lloyd DC (eds) Mutagen-induced chromosome damage in man. Edinburgh University Press, Edinburgh, pp 142–150
Cremer T, Lichter P, Borden J, Ward DC, Manuelidis L (1988a) Detection of chromosome aberrations in metaphase and interphase tumor cells by in situ hybridization using chromosome-specific library probes. Hum Genet 80:235–246
Cremer T, Tesin D, Hopman AHN, Manuelidis L (1988b) Rapid interphase and metaphase assessment of specific chromosomal changes in neuroectodermal tumor cell by in situ hybridization with chemically modified DNA probes. Exp Cell Res 176:199–220
Earnshaw WC, Ratrie H, Stetten G (1989) Visualization of centromere proteins CENP-B and CENP-C on a stable dicentric chromosome in cytological spreads. Chromosoma 98:1–12
Gray JW, Lucas JN, Yu LC, Langlois R (1984) Flow cytometric detection of aberrant chromosomes. In: Eisert WG, Mendelsohn ML (eds) Biological dosimetry. Springer, Berlin Heidelberg New York, pp 25–36
Green DK, Fantes JA (1987) Counting dicentric chromosomes by flow cytometry. In: Burger G, Ploem JS, Goerttler K (eds) Clinical cytometry and histometry. Academic Press, London, pp 263–268
ISCN (1985) An international system for human cytogenetic nomenclature. Harnden DG, Klinger HP, Jensen JT, Kaelbling M (eds) Karger, Basel, p 7
Jabs EW, Persico MG (1987) Characterization of human centromeric regions of specific chromosomes by means of alphoid DNA sequences. Am J Hum Genet 41: 374–390
Jabs EW, Persico MG (1989) Macromolecular organization of human centromeric regions reveals high-frequency, polymorphic macro DNA repeats. Proc Natl Acad Sci USA 86: 202–206
Johnson GD, deC Nogueira Araujo GM (1981) A simple method of reducing the fading of immunofluorescence during microscopy. J Immunol Methods 43:349–350
Koch JE, Kølvraa S, Petersen KP, Gregersen N, Bolund L (1989) Oligonucleotide-priming methods for the chromosome-specific labeling of alpha satellite DNA in situ. Chromosoma 98: 259–265
Landegent J, Jansen in de Wal N, Baan R, Hoeijmakers J, van derPloeg M (1984) 2-acetylaminofluorene-modified probes for the indirect hybridocytochemical detection of specific nucleic acid sequences. Exp Cell Res 153:61–72
Lloyd DC (1984) An overview of radiation dosimetry by conventional cytogenetic methods. In: Eisert WG, Mendelsohn ML (eds) Biological dosimetry. Springer, Berlin Heidelberg New York, pp 3–14
Lucas JN, Tenjin T, Straume T, Pinkel D, Moore D, Litt M, Gray JW (1989) Rapid human chromosome aberration analysis using fluorescence in situ hybridization. Int J Radiat Biol 56: 35–44
Lucas JN, Mullikin JC, Gray JW (1991) Dicentric chromosome frequency analysis using slit-scan flow cytometry. Cytometry 12:316–322
Maniatis T, Fritsch EF, Sambrook J (1986) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Manuelidis L (1978) Chromosomal localization of complex and simple repeated human DNAs. Chromosoma 66:23–32
Meyne J, Littlefield LG, Moyzis RK (1989) Labeling of human centromeres using an alphoid DNA consensus sequence: application to the scoring of chromosome aberrations. Mutat Res 226:75–79
Moroi Y, Peebles C, Fritzler MJ, Steigerwald J, Tan EM (1980) Autoantibody to centromere (kinetochore) in scleroderma sera. Proc Natl Acad Sci USA 77:1627–1631
Murray V, Martin RF (1987) Nucleotide sequences of human alpha-DNA repeats. Gene 57:255–259
Pinkel D, Gray JW, Trask B, van denEngh G, Fuscoe J, vanDekken H (1986) Cytogenetic analysis by in situ hybridization with fluorescently labeled nucleic acid probes. Cold Spring Harbor Symp Quant Biol LI: 151–157
Pinkel D, Landegent J, Collins C, Fuscoe J, Segraves R, Lucas J, Gray JW (1988) Fluorescence in situ hybridization with human chromosome-specific libraries: detection of trisomy 21 and translocations of chromosome 4. Proc Natl Acad Sci USA 85:9138–9142
Popp S, Cremer T, Lichter P, Cremer C (1989) A new approach to the rapid detection of radiation induced chromosome aberrations in human peripheral lymphocytes by in situ hybridization. Anal Cell Pathol 1:331
Saiki RK, Scharf S, Fallona F, Mullis KB, Horn GT, Erlich HA, Arnheim N (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
vanDekken H, Bauman JG (1988) A new application of in situ hybridization: detection of numerical and structural chromosome aberrations with a combination centromeric-telomeric probe. Cytogenet Cell Genet 48:188–189
Waye JS, Willard HF (1987) Nucleotide sequence heterogeneity of alpha satellite repetitive DNA: a survey of alphoid sequences from different human chromosomes. Nucleic Acids Res 15:7549–7569
Weier H-Ul, Eisert WG (1987) Quantification of dicentric chromosomes by dual parameter slit-scan cytometry. In: Burger G, Ploem JS, Goerttler K (eds) Clinical cytometry and histometry. Academic Press, London, pp 269–272
Weier H-Ul, Segraves R, Pinkel D, Gray JW (1990) Synthesis of Y chromosome-specific, labeled DNA probes. J Histochem Cytochem 38:421–426
Weier H-Ul, Kleine H-D, Gray JW (1991) Labeling of the centromeric region of human chromosomes 8 by in situ hybridization. Hum Genet (in press)
Willard HF Waye JS (1987) Hierarchical order in chromosome-specific human alpha satellite DNA. Trends Genet 3:192–198
Wu JC, Manuelidis L (1980) Sequence definition and organization of human repeated DNA. J Mol Biol 142: 363–386
Zinkowski RP, Vig BK, Broccoli D (1986) Characterization of kinetochores in multicentric chromosomes. Chromosoma 94:243–248
Author information
Authors and Affiliations
Additional information
by H.F. Willard
Rights and permissions
About this article
Cite this article
Weier, HU.G., Lucas, J.N., Poggensee, M. et al. Two-color hybridization with high complexity chromosome-specific probes and a degenerate alpha satellite probe DNA allows unambiguous discrimination between symmetrical and asymmetrical translocations. Chromosoma 100, 371–376 (1991). https://doi.org/10.1007/BF00337515
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00337515