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Detection of Loss of Heterozygosity in Tk-Deficient Mutants from L5178Y Tk+/−-3.7.2C Mouse Lymphoma Cells

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Molecular Toxicology Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2102))

Abstract

The mouse lymphoma assay (MLA), a forward mutation assay using the Tk+/−-3.7.2C clone of the L5178Y mouse lymphoma cell line and the Thymidine kinase (Tk) gene, has been widely used as an in vitro genetic toxicity assay for more than four decades. The MLA can evaluate the ability of mutagens to induce a wide range of genetic events including both gene mutations and chromosomal mutations and has been recommended as one component of several genotoxicity test batteries. Tk-deficient mutants often exhibit chromosomal abnormalities involving the distal end of chromosome 11 where the Tk gene is located, in mice, and the type of chromosome alteration can be analyzed using a loss of heterozygosity (LOH) approach. LOH has been considered an important event in human tumorigenesis and can result from any of the following several mechanisms: large deletions, mitotic recombination, and chromosome loss. In this chapter, the authors describe the procedures for the detection of LOH in the Tk mutants from the MLA, and apply LOH analysis for understanding the types of genetic damage that is induced by individual chemicals.

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Acknowledgments

We thank Drs. Tao Chen and Kelly Harris for their critical review of this manuscript. The information in this article is not a formal dissemination of information by the U.S. Food and Drug Administration (FDA) and does not represent Agency position or policy.

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Authors and Affiliations

  1. Division of Genetic and Molecular Toxicology, National Center for Toxicological Research, Jefferson, AR, USA

    Xiaoqing Guo & Ying Chen

  2. Ramboll, US Corporation, Little Rock, AR, USA

    Martha M. Moore

  3. Division of Genetic and Molecular Toxicology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR, USA

    Nan Mei

Authors
  1. Xiaoqing Guo
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  2. Ying Chen
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  3. Martha M. Moore
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  4. Nan Mei
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Correspondence to Nan Mei .

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Editors and Affiliations

  1. Department of Environmental and Occupational Health, Graduate School of Public Health, University of Pittsburgh, Pittsburgh, PA, USA

    Phouthone Keohavong

  2. Department of Environmental Toxicology, The Institute of Environmental and Human Health (TIEHH), Texas Tech University, Lubbock, TX, USA

    Kamaleshwar P. Singh

  3. Department of Occupational and Environmental Health Sciences, School of Public Health West Virginia University, Morgantown, WV, USA

    Weimin Gao

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Guo, X., Chen, Y., Moore, M.M., Mei, N. (2020). Detection of Loss of Heterozygosity in Tk-Deficient Mutants from L5178Y Tk+/−-3.7.2C Mouse Lymphoma Cells. In: Keohavong, P., Singh, K., Gao, W. (eds) Molecular Toxicology Protocols. Methods in Molecular Biology, vol 2102. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0223-2_14

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  • DOI: https://doi.org/10.1007/978-1-0716-0223-2_14

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