Skip to main content

Cytokinesis Blocked Micronuclei Aberration Analysis

  • Protocol
  • First Online:
Chromosome Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2519))

Abstract

Cytokinesis blocked micronuclei (CBMN) assay is a rapid and sensitive analysis of chromosome aberrations and miss assortments during cell division. Genotoxic agent exposure produces DNA damage and chromosome fragments. Fragmented chromosomes without centromere failed to attach kinetochore which segregates a pair of homologous chromosomes to each daughter cells at cytokinesis, hence leading to form micronuclei. Chromosome or fragments of chromosome can also form micronuclei when they are not accurately sorted to daughter cells. Using cytochalasin B, an actin inhibitor, blocks cytokinesis of which completion leads serration appearance formed with two daughter cells while nuclei segregation is undergoing. As a result, one cell having two daughter nuclei, i.e., binucleated cell, is produced. By analyzing these binucleated cells, chromosome aberrations can be estimated as well as popular chromosome aberration analysis. Frequency of micronuclei formation predicts the testing agents’ genotoxicity. By combining use with centromere-specific probes or DNA damage signal probes, the nature of genotoxicity of tested agents can be estimated.

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

Access this chapter

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

Similar content being viewed by others

References

  1. Engen A, Maeda J, Wozniak DE, Brents CA, Bell JJ, Uesaka M, Aizawa Y, Kato TA (2015) Induction of cytotoxic and genotoxic responses by natural and novel quercetin glycosides. Mutat Res Genet Toxicol Environ Mutagen 784–785:15–22. https://doi.org/10.1016/j.mrgentox.2015.04.007

    Article  CAS  PubMed  Google Scholar 

  2. Evans HJ, Neary GJ, Williamson FS (1959) The relative biological efficiency of single doses of fast neutrons and gamma-rays on Vicia faba roots and the effect of oxygen. Part II. Chromosone damage: the production of micronuclei. Int J Radiat Biol Relat Stud Phys Chem Med 1:216–229. https://doi.org/10.1080/09553005914550311

    Article  CAS  PubMed  Google Scholar 

  3. Heddle JA (1973) A rapid in vivo test for chromosomal damage. Mutat Res 18(2):187–190. https://doi.org/10.1016/0027-5107(73)90035-3

    Article  CAS  PubMed  Google Scholar 

  4. Countryman PI, Heddle JA (1976) The production of micronuclei from chromosome aberrations in irradiated cultures of human lymphocytes. Mutat Res 41(2–3):321–332. https://doi.org/10.1016/0027-5107(76)90105-6

    Article  CAS  PubMed  Google Scholar 

  5. Hayashi M, Sofuni T, Ishidate M Jr (1983) An application of Acridine Orange fluorescent staining to the micronucleus test. Mutat Res 120(4):241–247. https://doi.org/10.1016/0165-7992(83)90096-9

    Article  CAS  PubMed  Google Scholar 

  6. Fenech M, Morley AA (1985) Measurement of micronuclei in lymphocytes. Mutat Res 147(1–2):29–36

    Article  CAS  Google Scholar 

  7. Fenech M, Morley AA (1986) Cytokinesis-block micronucleus method in human lymphocytes: effect of in vivo ageing and low dose X-irradiation. Mutat Res 161(2):193–198. https://doi.org/10.1016/0027-5107(86)90010-2

    Article  CAS  PubMed  Google Scholar 

  8. Carter SB (1967) Effects of cytochalasins on mammalian cells. Nature 213(5073):261–264. https://doi.org/10.1038/213261a0

    Article  CAS  PubMed  Google Scholar 

  9. Brenner S, Pepper D, Berns MW, Tan E, Brinkley BR (1981) Kinetochore structure, duplication, and distribution in mammalian cells: analysis by human autoantibodies from scleroderma patients. J Cell Biol 91(1):95–102. https://doi.org/10.1083/jcb.91.1.95

    Article  CAS  PubMed  Google Scholar 

  10. Becker P, Scherthan H, Zankl H (1990) Use of a centromere-specific DNA probe (p82H) in nonisotopic in situ hybridization for classification of micronuclei. Genes Chromosomes Cancer 2(1):59–62. https://doi.org/10.1002/gcc.2870020111

    Article  CAS  PubMed  Google Scholar 

  11. Cartwright IM, Genet MD, Kato TA (2012) A simple and rapid fluorescence in situ hybridization microwave protocol for reliable dicentric chromosome analysis. J Radiat Res. https://doi.org/10.1093/jrr/rrs090

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Takamitsu A. Kato .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Kato, T.A. (2023). Cytokinesis Blocked Micronuclei Aberration Analysis. In: Gotoh, E. (eds) Chromosome Analysis. Methods in Molecular Biology, vol 2519. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2433-3_9

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-2433-3_9

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2432-6

  • Online ISBN: 978-1-0716-2433-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics