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

TY - JOUR

T1 - The application of the comet assay to assess the genotoxicity of environmental pollutants in the nematode Caenorhabditis elegans

AU - Imanikia, Soudabeh

AU - Galea, Francesca

AU - Nagy, Eszter

AU - Phillips, David H.

AU - Stürzenbaum, Stephen R.

AU - Arlt, Volker M.

PY - 2016/7

Y1 - 2016/7

N2 - 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 48 h exposure (0 to 40 μM). Induction of comets by BaP was concentration-dependent up to 20 μM; comet % tail DNA was ∼30% at 20 μM 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.

AB - 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 48 h exposure (0 to 40 μM). Induction of comets by BaP was concentration-dependent up to 20 μM; comet % tail DNA was ∼30% at 20 μM 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.

KW - single cell gel electrophoresis assay

KW - comet assay

KW - C. elegans

KW - benzo[a]pyrene

KW - DNA repair

UR - http://www.scopus.com/inward/record.url?scp=84977177573&partnerID=8YFLogxK

U2 - 10.1016/j.etap.2016.06.020

DO - 10.1016/j.etap.2016.06.020

M3 - Article

SN - 1382-6689

VL - 45

SP - 356

EP - 361

JO - ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY

JF - ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY

ER -