Single cell gel electrophoresis (SCGE) and Pig-a mutation assay in vivo-tools for genotoxicity testing from a regulatory perspective: A study of benzo[a]pyrene in Ogg1−/− mice

Graupner, Anne; Instanes, Christine; Dertinger, Stephen D.; Andersen, Jill Mari; Lindeman, Birgitte; Rongved, Tonje Danielsen; Brunborg, Gunnar; Olsen, Ann‐Karin

doi:10.1016/j.mrgentox.2014.07.010

Cited by 17 publications

(21 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…) (i) a time‐trend study measuring sperm DNA damage at different times after AA exposure and (ii) an obesity study analysing the combined effects of high‐fat diet (HFD) and GA exposure. For the time‐trend study, male C57BL/6 mice with BigBlue ™ background were used (bred at the Norwegian Institute of Public Health, as previously described; (Olsen et al ., ; Graupner et al ., ). For the obesity study, male C57BL/6NTac mice (Taconic, DK) were used.…”

Section: Methodsmentioning

confidence: 97%

Enhanced susceptibility of obese mice to glycidamide‐induced sperm chromatin damage without increased oxidative stress

et al. 2016

View full text Add to dashboard Cite

Diet-induced obesity is known to impair male reproduction and may aggravate the male reproductive toxicity of the food contaminant acrylamide. Exposure of male mice to acrylamide induces paternally mediated pre- and post-implantation losses because of spermatozoal toxicity and these effects are potentiated in mice fed a high-fat diet. Glycidamide - an acrylamide metabolite - is the primary mediator of reproductive effects in males. The mechanisms causing the interaction between diet and acrylamide are not clear. However, diet-induced obesity is associated with oxidative stress in male reproductive tissues which might contribute to increased germ cell susceptibility. In this study, we investigated whether a moderate diet-induced obesity regimen could interfere with glycidamide-induced spermatozoal toxicity and increase oxidative stress. For this purpose, sperm chromatin integrity, oxidised DNA and protein levels, transcript levels of oxidative stress responsive genes and glycidamide-induced DNA and haemoglobin adducts were analysed in samples from male mice exposed to a high-fat diet for 6 weeks in combination with a single glycidamide exposure 7 days prior to sacrifice. We found that glycidamide-induced sperm DNA fragmentation was markedly higher in obese than in lean mice. However, the levels of oxidised DNA and/or protein in blood, liver and testicular tissue was lower in obese than in lean mice. Accompanying the reduced level of oxidised macromolecules, the transcript levels of several oxidative stress-related genes were altered in the liver and testis from obese mice suggesting induction of an antioxidant response in these animals. The haemoglobin-glycidamide adduct levels were higher in obese than in lean animals, whereas obesity did not seem to increase the level of glycidamide-induced DNA adducts. These findings show that a moderate diet-induced obesity regimen may potentiate glycidamide-induced sperm cells toxicity and suggest that the increase in glycidamide-induced sperm toxicity observed in obese mice does not depend on overt oxidative stress.

show abstract

Section: Methodsmentioning

confidence: 97%

Enhanced susceptibility of obese mice to glycidamide‐induced sperm chromatin damage without increased oxidative stress

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Mutagenic effect of BaP was evident after different exposure scenarios in several mouse strains. This includes 28‐day dosing by oral gavage in the Muta™ mouse model , a single oral dose in Gpt delta mice , intraperitoneal injection for 3 days in C57BL/6 wild‐type and DNA repair‐deficient ( Ogg1 −/− ) mice and recently also after 3 days of exposure by oral gavage in CD‐1 mice . This demonstrates that for the prototypical mutagenic PAHs DMBA and BaP, the mouse Pig‐a gene mutation assay is able to reveal their genotoxic potential.…”

Section: Pig‐a In Micementioning

confidence: 94%

“…Several polycyclic aromatic hydrocarbons (PAHs) have been tested using the Pig‐a assay in mice . Both DMBA and the benzo[a]pyrene (BaP) showed clear induction of Pig‐a mutants. Mutagenic effect of BaP was evident after different exposure scenarios in several mouse strains.…”

Section: Pig‐a In Micementioning

confidence: 99%

“…Mice exhibiting different deficiencies in DNA repair proteins involved in protection of the genome from oxidized DNA have been studied [, Rolseth et al ., unpublished]. The DNA repair proteins investigated are involved in base excision repair eliminating oxidized DNA damage and comprise 8‐oxoguanine DNA glycosylase (Ogg1), the MutY DNA glycosylase (MutY) and three endonuclease VIII‐like DNA glycosylases (Neil1, Neil2 and Neil3) as well as the tumour protein p53 (p53).…”

Section: Usefulness In Basic Sciencementioning

confidence: 99%

See 1 more Smart Citation

The Pig‐a Gene Mutation Assay in Mice and Human Cells: A Review

Olsen

Dertinger

Krüger

et al. 2017

Basic Clin Pharma Tox

Self Cite

View full text Add to dashboard Cite

This MiniReview describes the principle of mutation assays based on the endogenous Pig-a gene and summarizes results for two species of toxicological interest, mice and human beings. The work summarized here largely avoids rat-based studies, as are summarized elsewhere. The Pig-a gene mutation assay has emerged as a valuable tool for quantifying in vivo and in vitro mutational events. The Pig-a locus is located at the X-chromosome, giving the advantage that one inactivated allele can give rise to a mutated phenotype, detectable by multicolour flow cytometry. For in vivo studies, only minute blood volumes are required, making it easily incorporated into ongoing studies or experiments with limited biological materials. Low blood volumes also allow individuals to serve as their own controls, providing temporal information of the mutagenic process, and/or outcome of intervention. These characteristics make it a promising exposure marker. To date, the Pig-a gene mutation assay has been most commonly performed in rats, while reports regarding its usefulness in other species are accumulating. Besides its applicability to in vivo studies, it holds promise for genotoxicity testing using cultured cells, as shown in recent studies. In addition to safety assessment roles, it is becoming a valuable tool in basic research to identify mutagenic effects of different interventions or to understand implications of various gene defects by investigating modified mouse models or cell systems. Human blood-based assays are also being developed that may be able to identify genotoxic environmental exposures, treatment- and lifestyle-related factors or endogenous host factors that contribute to mutagenesis.

show abstract

“…In vivo mutations in multiple organs can currently only be measured in transgenic rodents. The pigA assay has gained more attention during the last years as a potential alternative and the current developments with the PigA assay were presented .…”

mentioning

confidence: 99%

The European Environmental Mutagenesis and Genomics Society Annual Meeting, 14–18 August 2016, Copenhagen, Denmark

Holme

Frötschl

Knudsen

2017

Basic Clin Pharma Tox

View full text Add to dashboard Cite

The meeting was organized by EEMGS and ECOPA (The European Consensus Platform of Alternatives). It consisted of 10 sessions and a poster exhibition of 53 posters, 10 of which were by young scientists receiving grants from EEMGS, ECOPA, the 3R centre in Denmark and GUM. Together, there were four key note presentations, 50 oral presentations and 16 oral flash poster presentations. The number of participants was 180, from 18 countries (

show abstract

Single cell gel electrophoresis (SCGE) and Pig-a mutation assay in vivo-tools for genotoxicity testing from a regulatory perspective: A study of benzo[a]pyrene in Ogg1−/− mice

Cited by 17 publications

References 37 publications

Enhanced susceptibility of obese mice to glycidamide‐induced sperm chromatin damage without increased oxidative stress

Enhanced susceptibility of obese mice to glycidamide‐induced sperm chromatin damage without increased oxidative stress

The Pig‐a Gene Mutation Assay in Mice and Human Cells: A Review

The European Environmental Mutagenesis and Genomics Society Annual Meeting, 14–18 August 2016, Copenhagen, Denmark

Contact Info

Product

Resources

About