Detection of DNA-crosslinking agents with the alkaline comet assay

Pfuhler, Stefan; Wolf, Hans

doi:10.1002/(sici)1098-2280(1996)27:3<196::aid-em4>3.0.co;2-d

Cited by 175 publications

(68 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…In the single-cell gel assay, various compounds such as formaldehyde, cisplatinum or arsenite lead to decreasing tail moments with increasing concentrations, although these compounds are known to induce DNA lesions Pfuhler and Wolf 1996). This leads to false-negative results and can be overcome by the use of proteinase K in the lysis buer .…”

Section: Resultsmentioning

confidence: 99%

Genotoxicity of N -nitrosodicyclohexylamine in V79 cells in the sister chromatid exchange test and the single cell gel assay

et al. 2001

Archives of Toxicology

View full text Add to dashboard Cite

Dicyclohexylaminexnitrite is used in chemical formulations as an anti-corrosion agent. N-Nitrosodicyclohexylamine (N-NO-DCHA) can be formed by nitrosation from dicyclohexylamine during the application of these formulations. As most of the nitrosamines are genotoxic carcinogens, the genotoxic potential of N-NO-DCHA was investigated in V79 Chinese hamster cells in the single cell gel assay and the sister chromatid exchange (SCE) test. In addition, N-NO-DCHA cytotoxicity was determined in the neutral red assay. Neutral red uptake was suppressed up to 50% after 24 h incubation at a concentration of approximately 135 microM. In the single cell gel assay, a significantly elevated and dose-dependent induction of DNA lesions was detected in a concentration range from 5 microM to 100 microM (P<0.001). The use of proteinase K (1 mg/ml) in the lysing solution did not influence these results. In the SCE analysis, a significant induction of SCE was found at a minimum concentration of 5 microM N-NO-DCHA as well. A dose-dependent SCE induction could be detected up to the maximum concentration tested in the assay (100 microM). In conclusion, N-NO-DCHA is genotoxic in V79 cells in the single cell gel assay and the SCE test. With respect to human health hazard prevention, a substitution of dicyclohexylaminexnitrite in chemical formulations used to prevent corrosion is recommended.

show abstract

Section: Resultsmentioning

confidence: 99%

Genotoxicity of N -nitrosodicyclohexylamine in V79 cells in the sister chromatid exchange test and the single cell gel assay

et al. 2001

Archives of Toxicology

View full text Add to dashboard Cite

show abstract

“…A high innate activity of enzymes involved in rapid excision repair, however, can create high levels of incision-related breaks in the comet assay and therefore lead to a larger comet tail (Collins et al 1995). Cross-linked DNA, in contrast to other DNA damage, inhibits DNA migration in the electrophoretic field by stabilising the DNA (Merk and Speit 1999;Pfuhler and Wolf 1996).…”

Section: Assessing the Dna Damagementioning

confidence: 99%

The comet assay in male reproductive toxicology

2007

View full text Add to dashboard Cite

Due to our lifestyle and the environment we live in, we are constantly confronted with genotoxic or potentially genotoxic compounds. These toxins can cause DNA damage to our cells, leading to an increase in mutations. Sometimes such mutations could give rise to cancer in somatic cells. However, when germ cells are affected, then the damage could also have an effect on the next and successive generations. A rapid, sensitive and reliable method to detect DNA damage and assess the integrity of the genome within single cells is that of the comet or single-cell gel electrophoresis assay. The present communication gives an overview of the use of the comet assay utilising sperm or testicular cells in reproductive toxicology. This includes consideration of damage assessed by protocol modification, cryopreservation vs the use of fresh sperm, viability and statistics. It further focuses on in vivo and in vitro comet assay studies with sperm and a comparison of this assay with other assays measuring germ cell genotoxicity. As most of the de novo structural aberrations occur in sperm and spermatogenesis is functional from puberty to old age, whereas female germ cells are more complicated to obtain, the examination of male germ cells seems to be an easier and logical choice for research and testing in reproductive toxicology. In addition, the importance of such an assay for the paternal impact of genetic damage in offspring is undisputed. As there is a growing interest in the evaluation of genotoxins in male germ cells, the comet assay allows in vitro and in vivo assessments of various environmental and lifestyle genotoxins to be reliably determined.

show abstract

“…Under certain conditions, the assay can also detect DNA -DNA and DNA -protein crosslinking, which (in the absence of other kinds of DNA lesions) appears as a relative decrease in DNA migration compared to concurrent controls. In contrast to other DNA alterations, crosslinks may stabilize chromosomal DNA and inhibit DNA migration [70,71] . Thus, reduced DNA migration in comparison to the negative control (which should show some degree of DNA migration) may indicate the induction of crosslinks, which are relevant lesions with regard to mutagenesis and should be further investigated.…”

Section: Comet Assay For the Detection Of Dna Damagementioning

confidence: 99%