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Environ Toxicol Pharmacol. 2016 Jul;45:356-61. doi: 10.1016/j.etap.2016.06.020. Epub 2016 Jun 20.

The application of the comet assay to assess the genotoxicity of environmental pollutants in the nematode Caenorhabditis elegans.

Author information

  • 1Analytical and Environmental Sciences Division, MRC-PHE Centre for Environment and Health, King's College London, London, United Kingdom.
  • 2Analytical and Environmental Sciences Division, MRC-PHE Centre for Environment and Health, King's College London, London, United Kingdom; NIHR Health Protection Research Unit in Health Impact of Environmental Hazards at King's College London in partnership with Public Health England, London, United Kingdom.
  • 3Analytical and Environmental Sciences Division, MRC-PHE Centre for Environment and Health, King's College London, London, United Kingdom; NIHR Health Protection Research Unit in Health Impact of Environmental Hazards at King's College London in partnership with Public Health England, London, United Kingdom. Electronic address: volker.arlt@kcl.ac.uk.

Abstract

This study aimed to establish a protocol for cell dissociation from the nematode Caenorhabditis elegans (C. elegans) to assess the genotoxicity of the environmental pollutant benzo[a]pyrene (BaP) using the alkaline version of the single cell electrophoresis assay (comet assay). BaP genotoxicity was assessed in C. elegans (wild-type [WT]; N2, Bristol) after 48h exposure (0-40μM). Induction of comets by BaP was concentration-dependent up to 20μM; comet% tail DNA was ∼30% at 20μM BaP and ∼10% in controls. Similarly, BaP-induced DNA damage was evaluated in C. elegans mutant strains deficient in DNA repair. In xpa-1 and apn-1 mutants BaP-induced comet formation was diminished to WT background levels suggesting that the damage formed by BaP that is detected in the comet assay is not recognised in cells deficient in nucleotide and base excision repair, respectively. In summary, our study provides a protocol to evaluate DNA damage of environmental pollutants in whole nematodes using the comet assay.

KEYWORDS:

Benzo[a]pyrene; C. elegans; Comet assay; DNA repair; Single cell gel electrophoresis assay

PMID:
27389785
PMCID:
PMC4962771
DOI:
10.1016/j.etap.2016.06.020
[PubMed - indexed for MEDLINE]
Free PMC Article
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