Abstract
The rodent micronucleus assay has been most widely and frequently used as a representative in vivo assay system to assess mutagenicity of chemicals, regardless of endpoint of mutagenicity. The micronucleus has been developed to assess induction of structural and numerical chromosomal aberrations of target chemical. In this chapter, we describe the standard protocols of the assay using mouse bone marrow and peripheral blood. These methods are basically applicable to other rodents. The methodology of the micronucleus assay is rapidly developing, especially automatic analysis by flow cytometry (see also Chapter 11). Also we have to pay attention to the animal welfare, for example integration into repeat dose toxicity assay, combination of the micronucleus assay and Comet assay, and also omission of concurrent positive control group. Therefore, modification of the standard protocol is necessary for the actual assay on a case-by-case basis.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Heddle JA (1993) A rapid in vivo test for chromosomal damage. Mutat Res 18:187–190
Schmid W (1975) The micronucleus test. Mutat Res 31:9–15
Hayashi M, Sofuni T, Ishidate M Jr (1984) Kinetics of micronucleus formation in relation to chromosomal aberrations in mouse bone marrow. Mutat Res 127:129–137
Sutou S, Hayashi M, Nishi Y et al (1986) Sex difference in the micronucleus test. The collaborative study group for the micronucleus test. Mutat Res 172:151–163
Sutou S, Hayashi M, Shimada H et al (1988) Strain difference in the micronucleus test. The Collaborative Study Group for the Micronucleus Test. Mutat Res 204:307–316
Hayashi M, Sutou S, Shimada H et al (1989) Difference between intraperitoneal and oral gavage application in the micronucleus test: the 3rd collaborative study by CSGMT/JEMS.MMS. Mutat Res 223:329–344
Collaborative Study Group for the Micronucleus Test, the Mammalian Mutagenesis Study group of the Environmental Mutagen Society, Japan (CSGMT/JEMS⋅MMS) (1990) Single versus multiple dosing in the micronucleus test: the summary of the fourth collaborative study by CSGMT/JEMS⋅MMS. Mutat Res 234:205–222
The Collaborative Study Group for the Micronucleus Test (1992) Micronucleus test with mouse peripheral blood erythrocytes by acridine orange supravital staining: the summary report of the 5th collaborative study by CSGMT/JEMS—MMS. Mutat Res 278:83–98
Morita T, Asano N, Awogi T et al (1997) Evaluation of the rodent micronucleus assay in the screening of IARC carcinogens (Groups 1, 2A, and 2B). The summary report of the 6th collaborative study by CSGMT/JEMS⋅MMS. Mutat Res 389:3–122
Organization for Economic Co-operation and Development (OECD) (1997) OECD Guideline for the Testing of Chemicals 474 Mammalian Erythrocyte Micronucleus Test
International Conference on Harmonisation; guidance on S2(R1) (2011) Genotoxicity testing and data interpretation for pharmaceuticals intended for human use. http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Safety/S2_R1/Step4/S2R1_Step4.pdf
Hayashi M, Tice RR, MacGregor JT et al (1994) In vivo rodent erythrocyte micronucleus assay. Mutat Res 312:293–304
Hayashi M, MacGregor JT, Gatehouse DG et al (2000) In vivo rodent erythrocyte micronucleus assay. II. Some aspects of protocol design including repeated treatments, integration with toxicity testing, and automated scoring. Environ Mol Mutagen 35:234–252
Hayashi M, MacGregor JT, Gatehouse DG et al (2007) In vivo erythrocyte micronucleus assay III. Validation and regulatory acceptance of automated scoring and the use of rat peripheral blood reticulocytes, with discussion of non-hematopoietic target cells and a single dose-level limit test. Mutat Res 627:10–30
MacGregor JT, Wehr CM, Gould DH (1980) Clastogen-induced micronuclei in peripheral blood erythrocytes: the basis of an improved micronucleus test. Environ Mutagen 2:509–514
Hayashi M, Morita T, Kodama Y et al (1990) The micronucleus assay with mouse peripheral blood reticulocytes using acridine orange-coated slides. Mutat Res 245:245–249
Schlegel R, MacGregor JT (1984) The persistence of micronucleated erythrocytes in the peripheral circulation of normal and splenectomized Fischer 344 rats: implications for cytogenetic screening. Mutat Res 127:169–174
Hamada S, Sutou S, Morita T et al (2001) Evaluation of the rodent micronucleus assay by a 28-day-treatment protocol: summary of the 13th collaborative study by CSGMT/JEMS·MMS. Environ Mol Mutagen 37:93–110
Pfuhler S, Kirkland D, Kasper P et al (2009) Reduction of use of animals in regulatory genotoxicity testing: identification and implementation opportunities—report from an ECVAM workshop. Mutat Res 680:31–42
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, New York
About this protocol
Cite this protocol
Kasamoto, S., Masumori, S., Hayashi, M. (2013). In Vivo Micronucleus Assay in Mouse Bone Marrow and Peripheral Blood. In: Dhawan, A., Bajpayee, M. (eds) Genotoxicity Assessment. Methods in Molecular Biology, vol 1044. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-529-3_9
Download citation
DOI: https://doi.org/10.1007/978-1-62703-529-3_9
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-528-6
Online ISBN: 978-1-62703-529-3
eBook Packages: Springer Protocols