Abstract
Exposure to genotoxins may induce both structural and numerical chromosome aberrations. Whole chromosome fluorescence in situ hybridization (FISH) of metaphase cells provides a means for analyzing chromosome aberrations with single-cell level resolution. The laboratory mouse model is an essential tool for in vivo genotoxicity studies. However, few published resources are available that provide a comprehensive background and instructions on how to perform whole chromosome FISH on mouse metaphase cells. Here, we consolidate several techniques into a single, comprehensive protocol describing the preparation of mouse metaphase cells for whole chromosome FISH analysis. Also presented are basic recommendations on how to visualize the slides and how to organize the data for analysis and interpretation.
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References
Ceppi M, Gallo F, Bonassi S (2011) Study design and statistical analysis of data in human population studies with the micronucleus assay. Mutagenesis 26(1):247–252
Feneh M, Bonassi S (2011) The effect of age, gender, diet and lifestyle on DNA damage measured using micronucleus frequency in human peripheral blood lymphocytes. Mutagenesis 26(1):43–49
Mateuca R, Lombaert N, Aka PV et al (2006) Chromosomal changes: induction, detection methods and applicability in human biomonitoring. Biochimie 88(11):1515–1531
Ballarini F, Ottolenghi A (2004) Models of chromosome aberration induction: an example based on radiation track structure. Cytogenet Genome Res 104(1–4):149–156
Bailey SM, Bedford JS (2006) Studies on chromosome aberration induction: what can they tell us about DNA repair? DNA Repair (Amst) 5(9–10):1171–1181
Tucker JD, Marples B, Ramsey MJ et al (2004) Persistence of chromosome aberrations in mice acutely exposed to 56Fe+26 ions. Radiat Res 161(6):648–655
Breneman JW, Swiger RR, Ramsey MJ et al (1995) The development of painting probes for dual-color and multiple chromosome analysis in the mouse. Cytogenet Cell Genet 68(3–4):197–202
Tucker JD (2010) Chromosome translocations and assessing human exposure to adverse environmental agents. Environ Mol Mutagen 51(8–9):815–824
Tucker JD, Christensen ML (1987) Effects of anticoagulants upon sister-chromatid exchanges, cell-cycle kinetics, and mitotic index in human peripheral lymphocytes. Mutat Res 190(3):225–228
Delano MD (1995) Simple physical constraints in hemolysis. J Theor Biol 175(4): 517–524
Claussen U, Michel S, Mühlig P et al (2002) Demystifying chromosome preparation and the implications for the concept of chromosome condensation during mitosis. Cytogenet Genome Res 98(2–3):136–146
Deng W, Tsao SW, Lucas JN et al (2003) A new method for improving metaphase chromosome spreading. Cytometry A 51(1): 46–51
Henegariu O, Heerema NA, Lowe Wright L et al (2001) Improvements in cytogenetic slide preparation: controlled chromosome spreading, chemical aging and gradual denaturing. Cytometry 43(2):101–109
Petibone DM, Morris SM, Hotchkiss CE et al (2008) Technique for culturing Macaca mulatta peripheral blood lymphocytes for fluorescence in situ hybridization of whole chromosome paints. Mutat Res 653(1–2):76–81
Tucker JD, Morgan WF, Awa AA et al (1995) PAINT: a proposed nomenclature for structural aberrations detected by whole chromosome painting. Mutat Res 347(1):21–24
Tucker JD, Morgan WF, Awa AA et al (1995) A proposed system for scoring structural aberrations detected by chromosome painting. Cytogenet Cell Genet 68(3–4):211–221
Lucas JN, Tenjin T, Straume T et al (1989) Rapid human chromosome aberration analysis using fluorescence in situ hybridization. Int J Radiat Biol 56(1):35–44
Zhang L, Rothman N, Wang Y et al (1999) Benzene increases aneuploidy in the lymphocytes of exposed workers: a comparison of data obtained by fluorescence in situ hybridization in interphase and metaphase cells. Environ Mol Mutagen 34(4):260–268
Smith MT, Zhang L, Wang Y et al (1998) Increased translocations and aneusomy in chromosomes 8 and 21 among workers exposed to benzene. Cancer Res 58(10): 2176–2181
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The views presented in this article are those of the authors and do not necessarily reflect those of the U.S. Food and Drug Administration.
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Petibone, D.M., Tucker, J.D., Morris, S.M. (2014). Chromosome Painting of Mouse Peripheral Blood and Spleen Tissues. In: Sierra, L., Gaivão, I. (eds) Genotoxicity and DNA Repair. Methods in Pharmacology and Toxicology. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1068-7_8
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DOI: https://doi.org/10.1007/978-1-4939-1068-7_8
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1067-0
Online ISBN: 978-1-4939-1068-7
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