Skip to main content
SpringerLink
Log in

The In Vivo Rodent Micronucleus Test

  • Protocol
  • First Online:
Genotoxicity and DNA Repair

Part of the book series: Methods in Pharmacology and Toxicology ((MIPT))

Abstract

The in vivo mammalian micronucleus assay is the primary test in a battery of genotoxicity tests recommended by the regulatory agencies worldwide. The purpose of the assay is to identify substances that cause clastogenicity (chromosome breakage) and aneugenicity (chromosome lagging due to spindle dysfunction), and bone marrow toxicity by estimating the ratio of polychromatic erythrocytes to normochromatic erythrocytes. This chapter describes the mechanism of micronucleus formation, presents practical guidelines for designing studies, and gives the step-by-step protocols of the in vivo micronucleus test in bone marrow and peripheral blood cells of rodents.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Yunis JJ (1983) The chromosomal basis of human neoplasia. Science 221:227–236

    Article  CAS  PubMed  Google Scholar 

  2. Hagmar L, Bonassi S, Stromberg U et al (1998) Chromosomal aberration in lymphocytes predict human cancer: a report from the European Study Group on Cytogenetic Biomarkers and Health (ESCH). Cancer Res 58: 4117–4121

    CAS  PubMed  Google Scholar 

  3. Krishna G, Hayashi M (2000) In vivo rodent micronucleus assay: protocol, conduct and data interpretation. Mutat Res 455:155–166

    Article  CAS  PubMed  Google Scholar 

  4. Boller K, Schmid W (1970) Chemische Mutagenese beim Säunger. Das Knochenmark des Chinesischen Hamsters als in vivo-Testsystem. Hämatologische Befunde nach Behandlung mit Trenimon. Humangenetik II:35–54

    Google Scholar 

  5. Matter B, Schmid W (1971) Trenimon-induced chromosomal damage in bone marrow cells of six mammalian species, evaluated by the micronucleus test. Mutat Res 12:417–425

    Article  CAS  PubMed  Google Scholar 

  6. Schmid W, Arakaki DT, Breslau NA et al (1971) Chemical mutagenesis. The Chinese hamster bone marrow as an in vivo test system. I. Cytogenetic results on basic aspects of the methodology, obtained with alkylating agents. Humangenetk II:103–118

    Google Scholar 

  7. Schmid W (1973) Chemical mutagen testing on in vivo somatic mammalian cells. Agents Actions 3:77–85

    Article  CAS  PubMed  Google Scholar 

  8. Schmid W (1975) The micronucleus test. Mutat Res 31:9–15

    Article  CAS  PubMed  Google Scholar 

  9. Miller RC (1973) The micronucleus test as an in vivo cytogenetic method. Environ Health Perspect 6:167–170

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  10. Gollapudi BB, McFadden LG (1995) Sample size for the estimation of polychromatic to normochromatic erythrocyte ratio in the bone marrow micronucleus test. Mutat Res 347: 97–99

    Article  CAS  PubMed  Google Scholar 

  11. OECD (Organization for Economic Co-operation and Development) (1997) Mammalian Erythrocyte Micronucleus Test. Guideline for the testing of chemicals 474. Paris, France

    Google Scholar 

  12. Schlegel R, MacGregor JT (1984) The persistence of micronucleated erythrocytes in the peripheral circulation of normal and spenectomized Fischer 344 rats: implication for cytogenetic screening. Mutat Res 127:169–174

    Article  CAS  PubMed  Google Scholar 

  13. Abramsson-Zetterberg L, Grawé J, Zetterberg G (1999) The micronucleus test in rat erythrocytes from bone marrow, spleen and peripheral blood: the response to low doses of ionizing radiation, cyclophosphsmide and vincristine determined by flow cytometry. Mutat Res 423: 113–124

    Article  CAS  PubMed  Google Scholar 

  14. Gollapudi B, Kamra OP (1979) Application of a simple Giemsa-staining method in the micronucleus test. Mutat Res 64:45–46

    Article  CAS  PubMed  Google Scholar 

  15. 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

    Article  CAS  PubMed  Google Scholar 

  16. CSGMT (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.MSM. Mutat Res 278:83–98

    Article  Google Scholar 

  17. CSGMT (Collaborative Study Group for the Micronucleus Test) (1995) Protocol recommended for the short-term mouse peripheral blood micronucleus test. Mutagenesis 10: 153–159

    Article  Google Scholar 

  18. Hayashi M, Sofuni T, Ishidate-Jr M (1983) An application of acridine orange fluorescent staining to the micronucleus test. Mutat Res 120:241–247

    Article  CAS  PubMed  Google Scholar 

  19. Macgregor JT, Wehr CM, Langlois RG (1983) A simple fluorescent staining procedure for micronuclei and RNA in erythrocytes using Hoechst 33258 and Pyronin Y. Mutat Res 120:269–275

    Article  CAS  PubMed  Google Scholar 

  20. Krishna G, Fiedler R, Theiss JC (1992) Simultaneous evaluation of clastogenicity, aneugenicity, and toxicity in the mouse micronucleus assay using immunofluorescence. Mutat Res 282:159–304

    Article  CAS  PubMed  Google Scholar 

  21. Miller BM, Zitzelsberger HF, Weier HU et al (1991) Classification of micronuclei in mouse erythrocytes: immunofluorescent staining using CREST antibodies compared to in situ hybridization with biotinylated gamma satellite DNA. Mutagenesis 6:297–302

    Article  CAS  PubMed  Google Scholar 

  22. Margolin BH, Risko KJ (1998) The statistical analysis of in vivo genotoxicity data. Case studies of the rat hepatocyte UDS and mouse bone marrow micronucleus assay. In: Ashby J, deSerres FJ, Shelby MD, Margolin BH, Ishidate-Jr M, Becking (eds) Evaluation of short-term tests for carcinogens. Report of the international programme on chemical safety’s collaborative study on in vivo assay. Oxford University Press, Oxford, pp 29–44

    Google Scholar 

  23. Richold M, Ashby J, Bootman J et al (1990) In vivo cytogenetic assays. In: Kirkland DJ (ed) Basic mutagenicity tests, UKEMS recommended procedures. UKEMS subcommittee on guidelines for mutagenicity testing. Report. Part I revised. Cambridge University Press, Cambridge, pp 115–141

    Google Scholar 

  24. Lovell DP, Anderson D, Albanese R et al (1989) Statistical analysis of in vivo cytogenetic assays. In: Kirkland DJ (ed) Statistical evaluation of mutagenicity test data. UKEMS subcommittee on guidelines for mutagenicity testing. Report. Part III. Cambridge University Press, Cambridge, pp 184–232

    Google Scholar 

Download references

Acknowledgements

I am grateful to Eduardo de Souza Marques for his help in preparing the micronucleus origin schema shown in Fig. 1.

Author information

Authors and Affiliations

  1. Departamento de Fonoaudiologia, Universidade Estadual Paulista—UNESP—Faculdade de Filosofia e Ciências, Marilia, SP, 17525-900, Brazil

    Edson Luis Maistro

Authors
  1. Edson Luis Maistro
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Edson Luis Maistro .

Editor information

Editors and Affiliations

  1. Universidad de Oviedo, Oviedo, Spain

    L. María Sierra

  2. Universidade de Trás-os-Montes e Alto Douro, Vila Real, Portugal

    Isabel Gaivão

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media New York

About this protocol

Cite this protocol

Maistro, E.L. (2014). The In Vivo Rodent Micronucleus Test. 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_6

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-1068-7_6

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1067-0

  • Online ISBN: 978-1-4939-1068-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation