Nature Genetics
14, 312 - 315 (1996)
doi:10.1038/ng1196-312
Multicolour spectral karyotyping of mouse chromosomesMarek Liyanage1, Allen Coleman2, Stan du Manoir1, Tim Veldman1, Stephen McCormack3, Robert B. Dickson3, Carrolee Barlow4, Anthony Wynshaw-Boris4, Siegfried Janz2, Johannes Wienberg5, Malcolm A. Ferguson-Smith5, Evelin Schröck1
& Thomas Ried1, 6
1Diagnostic Development Branch, National Center for Human Genome Research, National Institutes of Health, Bldg. 49, 49 Convent Drive, Rm. 4A28, Bethesda, Maryland 20892-4470, USA
2Laboratory of Genetics, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA
3Lombardi Cancer Center, Georgetown University, Washington, D.C., USA
4Laboratory of Genetic Disease Research, National Center for Human Genome Research, National Institutes of Health, Bldg. 49, 49 Convent Drive, Rm. 4A28, Bethesda, Maryland 20892-4470, USA
5Department of Pathology, Cambridge University, Cambridge, UK
6e-mail: tried@nchgr.nih.gov Murine models of human carcinogenesis are exceedingly valuable tools to understand genetic mechanisms of neoplastic growth. The identification of recurrent chromosomal rearrangements by cytogenetic techniques serves as an initial screening test for tumour specific aberrations. In murine models of human carcinogenesis, however, karyotype analysis is technically demanding because mouse chromosomes are acrocentric and of similar size. Fluorescence in situ hybridization (FISH) with mouse chromosome specific painting probes1 can complement conventional banding analysis. Although sensitive and specific, FISH analyses are restricted to the visualization of only a few mouse chromosomes at a time. Here we apply a novel imaging technique2 that we developed recently for the visualization of human chromosomes3 to the simultaneous discernment of all mouse chromosomes. The approach is based on spectral imaging to measure chromosome-specific spectra after FISH with differentially labelled mouse chromosome painting probes. Utilizing a combination of Fourier spectroscopy, CCD-imaging and conventional optical microscopy, spectral imaging allows simultaneous measurement of the fluorescence emission spectrum at all sample points. A spectrum-based classification algorithm has been adapted to karyotype mouse chromosomes. We have applied spectral karyotyping (SKY) to chemically induced plasmocytomas, mammary gland tumours from transgenic mice overexpressing the c-myc onco-gene and thymomas from mice deficient for the ataxia telangiectasia (Atm) gene. Results from these analyses demonstrate the potential of SKY to identify complex chromosomal aberrations in mouse models of human carcinogenesis.
REFERENCES
- Rabbitts, P. et al. Chromosome specific paints from a high resolution flow karyotype of the mouse. Nature Genet. 9, 369−375 (1995). | Article | PubMed | ISI | ChemPort |
- Malik, Z. et al. Fourier transform muitipixel spectroscopy for quantitative cytology. J. Microscopy 182, 133−140 (1996). | Article | ISI | ChemPort |
- Schröck, E. et al. Multicolor spectral karyotyping of human chromosomes. Science 273, 494−497 (1996). | PubMed | ISI |
- Speicher, M.R., Ballard, S.G. & Ward, D.C. Karyotyping human chromosome by combinatorial multi-fluor FISH. Nature Genet. 12, 368−375 (1996). | Article | PubMed | ISI | ChemPort |
- Kastan, M. Experimental models of human carcinogenesis. Nature Genet. 5, 207−208 (1993). | PubMed | ISI | ChemPort |
- Potter, M. & Wiener, F. Piasmocytomagenesis in mice: model of neoplastic development dependent upon chromosomal translocations. Carcinogenesis 13, 1681−1697 (1992). | PubMed | ISI | ChemPort |
- Fearon, E.R. & Vogelstein, B. A genetic model for colorectal carcinogenesis. Cell 61, 759−767 (1990). | Article | PubMed | ISI | ChemPort |
- Bieche, I. & Lidereau, R. Genetic alterations in breast cancer. Genes Chrom. Cancer 4, 227−251 (1995).
- Cardiff, R.D. & Muller, W.J. Transgenic models of mammary tumorigenesis. Cancer Surveys 16, 97−113 (1993). | PubMed | ISI | ChemPort |
- Leder, A., Pattengale, P.K., Kuo, A., Stewart, T. & Leder, P. Consequences of widespread deregulation of the c-myc gene in transgenic mice: multiple neoplasms and normal development. Cell 45, 485−495 (1986). | Article | PubMed | ISI | ChemPort |
- Amundadottir, L.T. et al. TGF
 and c-myc cooperation in mouse mammary tumorigenesis: coordinate stimulation of growth and suppression of apoptosis. Oncogene 13, 757−765 (1996). | PubMed | ISI | ChemPort |
- Capecchi, M.R. Altering the genome by homologous recombination. Science 244, 1288−1292 (1989). | PubMed | ISI | ChemPort |
- Barlow, C. et al. Atm-deficient mice:a paradigm of ataxia telangiectasia. Cell 86, 159−171 (1996). | PubMed | ISI | ChemPort |
- Taylor, A.M., Metcalfe, J.A., Thick, J. & Mak, Y.F. Leukemia and lymphoma in ataxia telangiectasia. Blood 87, 423−438 (1996). | PubMed | ISI | ChemPort |
- Telenius, H. et al. Cytogenetic analysis by chromosome painting using DOP-PCR amplified flow sorted chromosomes. Genes Chrom. Cancer 4, 267−263 (1992). | PubMed |
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