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Sister Chromatid Exchange as a Genotoxic Stress Marker

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Radiation Cytogenetics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1984))

Abstract

Sister chromatid exchange (SCE) is the phenomenon of partial DNA exchange during DNA replication. SCE detection has been developed through eliciting DNA’s semiconservative replicative nature. Thymidine analogues such as 5′-bromodeoxyuridine (BrdU) and ethynyldeoxyuridine (EdU) are incorporated into the newly synthesized DNA for two cell cycles. The addition of Colcemid to the culture blocks and synchronizes cells at mitosis, and conventional cytogenetic preparations are made. Differential staining methods with Hoechst dye and Giemsa (Fluorescence Plus Giemsa staining), antibody detection against BrdU, or highly specific Click reaction to EdU, allow the newly synthesized DNA within a chromatid to be recognized. SCEs represent a point of DNA template exchange during DNA synthesis, visualized by differential chromatid staining or harlequin chromosomes. We will introduce three basic protocols in this chapter including non-fluorescence and fluorescence methods for SCE microscopic analysis. SCE is a very sensitive marker of genotoxic stress during replication.

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References

  1. Taylor JH, Woods PS, Hughes WL (1957) The organization and duplication of chromosomes as revealed by autoradiographic studies using tritium-labeled thymidinee. Proc Natl Acad Sci U S A 43:122–128

    Article  CAS  Google Scholar 

  2. Gebhart E (1981) Sister chromatid exchange (SCE) and structural chromosome aberration in mutagenicity testing. Hum Genet 58:235–254

    Article  CAS  Google Scholar 

  3. Nagasawa H, Little JB (1992) Induction of sister chromatid exchanges by extremely low doses of alpha-particles. Cancer Res 52:6394–6396

    CAS  PubMed  Google Scholar 

  4. Ellis NA, Proytcheva M, Sanz MM, Ye TZ, German J (1999) Transfection of BLM into cultured bloom syndrome cells reduces the sister-chromatid exchange rate toward normal. Am J Hum Genet 65:1368–1374

    Article  CAS  Google Scholar 

  5. Wilson DM III, Thompson LH (2007) Molecular mechanisms of sister-chromatid exchange. MutatRes 616:11–23

    CAS  Google Scholar 

  6. Conrad S, Kunzel J, Lobrich M (2011) Sister chromatid exchanges occur in G2-irradiated cells. Cell Cycle 10:222–228

    Article  CAS  Google Scholar 

  7. Gutierrez C, Gonzalez-Gil G, Hernandez P (1983) Analysis of baseline and BrdU-dependent SCEs at different BrdU concentrations. Exp Cell Res 149:461–469

    Article  CAS  Google Scholar 

  8. Heartlein MW, O’Neill JP, Preston RJ (1983) SCE induction is proportional to substitution in DNA for thymidine by CldU and BrdU. Mutat Res 107:103–109

    Article  CAS  Google Scholar 

  9. Suzuki H, Yosida TH (1983) Frequency of sister-chromatid exchanges depending on the amount of 5-bromodeoxyuridine incorporated into parental DNA. Mutat Res 111:277–282

    Article  CAS  Google Scholar 

  10. Hazen MJ, Villanueva A, Juarranz A, Canete M, Stockert JC (1985) Photosensitizing dyes and fluorochromes as substitutes for 33258 Hoechst in the fluorescence-plus-Giemsa (FPG) chromosome technique. Histochemistry 83:241–244

    Article  CAS  Google Scholar 

  11. Nagasawa H, Little JB (1979) Effect of tumor promoters, protease inhibitors, and repair processes on X-ray-induced sister chromatid exchanges in mouse cells. Proc Natl Acad Sci U S A 76:1943–1947

    Article  CAS  Google Scholar 

  12. Su C, Allum AJ, Aizawa Y, Kato TA (2016) Novel glyceryl glucoside is a low toxic alternative for cryopreservation agent. Biochem Biophys Res Commun 476:359–364

    Article  CAS  Google Scholar 

  13. Cartwright IM, Kato TA (2015) Role of various DNA repair pathways in chromosomal inversion formation in CHO mutants. Int J Radiat Biol 91:925–933

    Article  CAS  Google Scholar 

  14. Nagasawa H, Fornace D, Little JB (1983) Induction of sister-chromatid exchanges by DNA-damaging agents and 12-O-tetradecanoyl-phorbol-13-acetate (TPA) in synchronous Chinese hamster ovary (CHO) cells. Mutat Res 107:315–327

    Article  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. Department of Molecular Genetics, Medical Research Institute, Tokyo Medical and Dental University, Tokyo, Japan

    Shigeaki Sunada

  2. Department of Environmental and Radiological Health Sciences, Colorado State University, Fort Collins, CO, USA

    Jeremy S. Haskins & Takamitsu A. Kato

Authors
  1. Shigeaki Sunada
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  2. Jeremy S. Haskins
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  3. Takamitsu A. Kato
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Corresponding author

Correspondence to Shigeaki Sunada .

Editor information

Editors and Affiliations

  1. Department of Environmental and Radiological Health Sciences, Colorado State University, Fort Collins, CO, USA

    Takamitsu A. Kato

  2. Department of Radiation Oncology, University of California Davis, Sacramento, CA, USA

    Paul F. Wilson

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Sunada, S., Haskins, J.S., Kato, T.A. (2019). Sister Chromatid Exchange as a Genotoxic Stress Marker. In: Kato, T., Wilson, P. (eds) Radiation Cytogenetics. Methods in Molecular Biology, vol 1984. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9432-8_7

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  • DOI: https://doi.org/10.1007/978-1-4939-9432-8_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9430-4

  • Online ISBN: 978-1-4939-9432-8

  • eBook Packages: Springer Protocols

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