The mouse peripheral blood micronucleus test with 2-acetylaminofluorene using the acridine orange supravital staining method |
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| Affiliation: | 1. Toxicological Research Division, Nitto Denko Corporation, 1-1-2 Shimohozumi, Ibaraki, Osaka 567 Japan;2. Laboratory Animal Science and Toxicology Laboratories, Sankyo Co., Ltd., 717 Horikoshi, Fukuroi, Shizuoka 437 Japan;1. Department of Food and Nutrition, Providence University, Taichung, Taiwan, Republic of China;2. Department of Medical Laboratory Science and Biotechnology, Central Taiwan University of Sciences and Technology Taichung, Taiwan, Republic of China;3. Graduate Institute of Veterinary Pathology, National Chung Hsing University, Taichung, Taiwan, Republic of China;1. Institute of Cancer Research, Department of Medicine I, Medical University of Vienna, Vienna, Austria;2. Center for Public Health, Department of Environmental Health, Medical University of Vienna, Vienna, Austria;3. School of Pharmacy and Medical Sciences, University of South Australia, Adelaide, Australia;4. Universiti Kebangsaan Malaysia, Selangor, Malaysia;5. Genome Health Foundation, North Brighton, SA, Australia;6. Institute of Pharmacology and Toxicology, Wuerzburg University, Wuerzburg, Germany;7. Laboratory of Genetic Toxicology, Lutheran University of Brazil (ULBRA) & LaSalle University (UniLaSalle), Canoas, RS, Brazil;8. Environmental Carcinogenesis Unit, Ospedale Policlinico San Martino, Genoa, Italy |
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| Abstract: | The peripheral blood micronucleus test using the acridine orange (AO) supravital staining method was validated with the potent bone marrow clastogen 2-acetylaminofluorene (2-AAF). 2-AAF induced micronuclei in peripheral blood reticuiocytes dose-dependently as well as in bone marrow polychromatic erythrocytes. The incidence of micronucleated reticuiocytes (MNRETs) peaked 48 h after a single treatment in both CD-1 and BDF1 mice, and the incidence of micronucleated polychromatic erythrocytes (MNPCEs) peaked 24 or 48 h after treatment. The maximum incidences of MNRETs were always higher than those of MNPCEs in both mouse strains treated once. In the double-treatment regime, the maximum incidence of MNRETs was observed at 24 h after the second treatment in each strain. The incidences of MNRETs in BDF1 mice were higher than in CD-1 mice after a single treatment but were comparable after double treatment.These results indicate that the peripheral blood micronucleus test using AO supravital staining is as sensitive as the conventional bone marrow assay. The new staining method can be performed more easily than the original smear method using either bone marrow or peripheral blood cells. Thus, the peripheral blood method using AO supravital staining is a possible alternative to the conventional bone marrow assay. |
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