Skip to main content

The Comet Assay: A Reliable and Sensitive Method for the Documentation of UV-B Induced DNA Damage

  • Chapter
Ocular Toxicology

Abstract

Evidence exists that ultraviolet radiation, drugs, chemicals, and inherent mechanisms directly or via the generation of inflammatory mediators and oxygen free radicals can exert effects on DNA damage and repair1,2,3. Thus the evaluation of DNA damage and the investigation of factors influencing its quantification are of prime interest for pharmaceutical, clinical and basic scientific reasons4,5,6. Previous approaches towards the quantification of DNA damage in lens epithelial cells lacked a systematic analysis of individual factors such as cell origin, cell cycle phase, Go synchronization, and the interaction between these factors.

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

Access this chapter

eBook
USD 16.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 16.99
Price excludes VAT (USA)
  • Compact, lightweight 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

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Andley, U.P., Lewis, R.M., Reddan, J.R. Kochevar, I.R. Action spectrum for cytotoxicicty in the UV-A-and UV-B wavelength region in cultured lens epithelial cells, Invest. Ophthalmol. Vis. Sci. 35:367–373 (1994).

    PubMed  CAS  Google Scholar 

  2. M.V. Riley S. Susan, M.J. Peters, C.A. Schwartz, The effects of UV-B irradiation on the corneal endothelium, Current Eye Research 6:1021–1033 (1987).

    Article  PubMed  CAS  Google Scholar 

  3. M.V. Riley, S.A. Elgebaly, The release of a neutrophil chemotactic factor from UV-B irradiated rabbit corneas, Current Eye Research 9:677–682 (1990).

    Article  PubMed  CAS  Google Scholar 

  4. L.A. Applegate, R.D. Ley, DNA damage is involved in the induction of opacification and neovascularization of the cornea by ultraviolet radiation, Exp. Eye Res. 52:493–497 (1991).

    Article  PubMed  CAS  Google Scholar 

  5. N.J. Kleiman, A. Spector, DNA single strand breaks in human lens epithelial cells from patients with cataract, Current Eye Res. 12:423–431 (1990).

    Article  Google Scholar 

  6. A. Spector, G.-M. Wang, R.-R. Wang, The prevention of cataract caused by oxidative stress in cultured rat lenses. II. Early effects of photochemical stress and recovery, Exp. Eye Res. 57:659–667 (1993).

    Article  PubMed  CAS  Google Scholar 

  7. N.P. Singh, M.T. McCoy, R.R. Tice, E.L. Schneider, A simple technique for quantification of low levels of DNA damage in individual cells, Exp. Cell Res. 175:184–191 (1988).

    Article  PubMed  CAS  Google Scholar 

  8. P.L. Olive, D. Wlodek, R.E. Durand, J.P. Banáth, Factors influencing DNA migration from individual cells subjected to gel electrophoresis, Exp. Cell Res. 198:259–287 (1992).

    Article  PubMed  CAS  Google Scholar 

  9. V.J. McKelvey-Martin, M.H.L. Green, P. Schmezer, B.L. Pool-Zobel, M.P. De Meo, A. Collins, The single cell gel electrophoresis assay (comet assay): a European review, Mutation Res. 288:47–63 (1993).

    Article  PubMed  CAS  Google Scholar 

  10. W. Breipohl, M. Leyendecker, O. Leip, H. Akiyoshi, Investigations on cell proliferation of bovine lens epithelial cells, submitted (1995).

    Google Scholar 

  11. O. Leip, W. Breipohl, H. Akiyoshi, T. Naguro, M. Leyendecker, C. Penzkofer, C., Effects of UV-B radiation on cultured bovine lens epithelial cells, Abstract, XXVII Intern. Congr. Ophthalmol. Toronto (1994).

    Google Scholar 

  12. A.J. Fornace, Mammalian genes induced by radiation; activation of genes associated with growth control,Annu. Rev. Genet. 26:507–526 (1992).

    Article  PubMed  CAS  Google Scholar 

  13. M. Molls, C. Streffer, The influence of G2 progression on X-ray sensitivity of two cell mouse embryos,Int. J. Radiat. Biol. Relat. Stud. Phys. Chem Med. 46:355–365 (1984).

    Article  PubMed  CAS  Google Scholar 

  14. R. deJong, J.W. McAvoy, Distribution of acidic and basic fibroblast growth factors (FGF) in the foetal rat eye: implications for lens development, Growth Factors 6:159–177 (1992).

    Google Scholar 

  15. T. Kartasova, P.v.d. Putte, Isolation, characterization, and UV-stimulated expression of two families of genes encoding polypeptides of related structure in human epidermal keratinocytes, Mol. Cell. Biol. 8:2195–2203 (1988).

    PubMed  CAS  Google Scholar 

  16. Q. Zhan, K.A. Lord, I. Alamo jr., M. C. Hollander, F. Carrier, D. Ron, K. W. Kohn, B. Hoffman,D.A. Liebermann, A.J. Fornace jr., The gadd and Myd genes define a novel set of mammalian genes encoding acidic proteins that synergistically suppress cell growth, Mol Cell. Biol. 14:2361–2371 (1994).

    Article  PubMed  CAS  Google Scholar 

  17. Q. Zhan, F. Carrier, A.J. Fornace jr., Induction of cellular p53 activity by DNA-damaging agents and growth arrest, Mol. Cell. Biol. 13:4242–4250 (1993).

    PubMed  CAS  Google Scholar 

  18. A.J. Fornace jr., D.W. Nebert, M.C. Hollander, J.D. Luethy, M. Papathanasiou, J. Fargnoli, N.J.Holbrook, Mammalian genes coordinately regulated by growth arrest signals and DNA-damaging agents, Mol. Cell Biol. 9:4196–4203 (1989).

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1995 Springer Science+Business Media New York

About this chapter

Cite this chapter

Breipohl, W. et al. (1995). The Comet Assay: A Reliable and Sensitive Method for the Documentation of UV-B Induced DNA Damage. In: Weisse, I., Hockwin, O., Green, K., Tripathi, R.C. (eds) Ocular Toxicology. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-1887-7_40

Download citation

  • DOI: https://doi.org/10.1007/978-1-4615-1887-7_40

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-5769-8

  • Online ISBN: 978-1-4615-1887-7

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics