Flow cytometric analysis of micronuclei in rat peripheral blood: An interlaboratory reproducibility study

Kasamoto, Sawako; Mukai, Daisuke; Masumori, Shoji; Suzuki, Kazutoshi; Tanaka, Ryota; Torous, Dorothea K.; Yamate, Jyoji; Hayashi, Makoto

doi:10.1016/j.mrgentox.2014.02.001

Cited by 3 publications

(1 citation statement)

References 13 publications

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“…The mammalian erythrocyte micronucleus test is the primary regulatory assay for the assessment of chromosomal damage in vivo (OECD, 1997). In addition to scoring micronuclei on slides using a microscope, the flow cytometry-based automated analysis method for the enumeration of micronuclei in peripheral blood reticulocytes has been validated over decades of use and was adopted by OECD in 2014 Torous et al, 2005;Dertinger et al, 2006;Kasamoto et al, 2014;OECD, 2014).…”

Section: Introductionmentioning

confidence: 99%

Assessment of the Pig‐a, micronucleus, and comet assay endpoints in rats treated by acute or repeated dosing protocols with procarbazine hydrochloride and ethyl carbamate

Gaofeng

Wen

Zhihui

et al. 2018

Environ and Mol Mutagen

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The utility and sensitivity of the newly developed flow cytometric Pig‐a gene mutation assay have become a great concern recently. In this study, we have examined the feasibility of integrating the Pig‐a assay as well as micronucleus and Comet endpoints into acute and subchronic general toxicology studies. Male Sprague–Dawley rats were treated for 3 or 28 consecutive days by oral gavage with procarbazine hydrochloride (PCZ) or ethyl carbamate (EC) up to the maximum tolerated dose. The induction of CD59‐negative reticulocytes and erythrocytes, micronucleated reticulocytes in peripheral blood, micronucleated polychromatic erythrocytes in bone marrow, and Comet responses in peripheral blood, liver, kidney, and lung were evaluated at one, two, or more timepoints. Both PCZ and EC produced positive responses at most analyzed timepoints in all tissue types, both with the 3‐day and 28‐day treatment regimens. Furthermore, comparison of the magnitude of the genotoxicity responses indicated that the micronucleus and Comet endpoints generally produced greater responses with the higher dose, short‐term treatments in the 3‐day study, while the Pig‐a assay responded better to the cumulative effects of the lower dose, but repeated subchronic dosing in the 28‐day study. Collectively, these results indicate that integration of several in vivo genotoxicity endpoints into a single routine toxicology study is feasible and that the Pig‐a assay may be particularly suitable for integration into subchronic dose studies based on its ability to accumulate the mutations that result from repeated treatments. This characteristic may be especially important for assaying lower doses of relatively weak genotoxicants. Environ. Mol. Mutagen. 60:56–71, 2019. © 2018 Wiley Periodicals, Inc.

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Section: Introductionmentioning

confidence: 99%

Assessment of the Pig‐a, micronucleus, and comet assay endpoints in rats treated by acute or repeated dosing protocols with procarbazine hydrochloride and ethyl carbamate

Gaofeng

Wen

Zhihui

et al. 2018

Environ and Mol Mutagen

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Increased micronucleus frequencies in reticulocytes of children exposed to industrial pollution: oxidative stress and the OGG1 S326C polymorphism

Montero‐Montoya

López-Vargas

Méndez-Serrano³

et al. 2020

Mutation Research/Genetic Toxicology and Environmental Mutagene

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The OECD’s micronucleus test guideline for single exposure to an agent and the genotox-kinetic alternative

Morales-Ramı́rez¹,

Vallarino-Kelly²,

Cruz-Vallejo³

2017

Mutagenesis

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The 'Organization for Economic Co-operation and Development (OECD) guidelines for the Testing of Chemicals' aims to identify whether a chemical is a genotoxic hazard, and these guidelines 'are periodically reviewed in the light of scientific progress, changing regulatory needs and animal welfare considerations'. OECD published a mammalian erythrocyte micronucleus test guideline for testing chemicals (1) that proposes: 'Animals are treated with the test chemical once…Samples of peripheral blood are taken at least twice (from the same group of animals), starting not earlier than 36 h after treatment, with appropriate intervals following the first sample, but not extending beyond 72 h'. This guidelines are base on the report by the Collaborative Study Group for the Micronucleus Test (CSGMT), which was based on the sampling of mice peripheral blood every 24 h We investigated the kinetics of micronucleus induction by taking samples prior to administration and every 8 or 10 h after single treatment. The comparisons suggest that 24-h sampling could cause not only an underestimation of the responses to various agents but also a misestimation of the time of maximal induction. We proposed that samples of peripheral blood must be collected at two different times during an optimal 25-h sampling range (from 25 to 50 h). Besides, we hypothesize that the time of maximal EPC-MN induction depends on the time required for the mechanisms involved in micronucleus production; and we suggest that a kinetic analysis of MN-PCE induction by several agents with well-known mechanisms of micronuclei induction would allow derivation of a specific relationship between the kinetics of MN-PCE induction and some process of DNA breaks and/or micronuclei induction.

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Flow cytometric analysis of micronuclei in rat peripheral blood: An interlaboratory reproducibility study

Cited by 3 publications

References 13 publications

Assessment of the Pig‐a, micronucleus, and comet assay endpoints in rats treated by acute or repeated dosing protocols with procarbazine hydrochloride and ethyl carbamate

Assessment of the Pig‐a, micronucleus, and comet assay endpoints in rats treated by acute or repeated dosing protocols with procarbazine hydrochloride and ethyl carbamate

Increased micronucleus frequencies in reticulocytes of children exposed to industrial pollution: oxidative stress and the OGG1 S326C polymorphism

The OECD’s micronucleus test guideline for single exposure to an agent and the genotox-kinetic alternative

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