The micronucleus assay using peripheral blood reticulocytes from mitomycin C- and cyclophosphamide-treated rats |
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| Affiliation: | 1. Division of Genetics and Mutagenesis, National Institute of Hygienic Sciences 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158 Japan;2. Division of Toxicology, National Institute of Hygienic Sciences 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158 Japan;3. Otsuka Pharmaceutical Co., Ltd., 463-10, Kagasuno, Kawauchi-cho, Tokushima 771-01 Japan;1. Department of Industrial Engineering, CEG Campus, Anna University, Chennai 600025, India;2. Department of Mechanical Engineering, SSN College of Engineering, Chennai 603110, India;1. Department of Pathology, The Johns Hopkins University School of Medicine, Baltimore, Maryland;2. Department of Pathology, University of Texas MD Anderson Cancer Center, Houston, Texas;3. Department of Pathology and Laboratory Medicine, Weil Cornell Medicine, New York, New York;4. Department of Radiology, The Johns Hopkins University School of Medicine, Baltimore, Maryland;1. Laboratory of Nano- and Translational Medicine, Department of Radiation Oncology, Lineberger Comprehensive Cancer Center, Carolina Center for Cancer Nanotechnology Excellence, Carolina Institute of Nanomedicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina;2. Division of Medical Oncology, Department of Medicine, Duke Cancer Institute, Duke University Medical Center, Durham, North Carolina;3. Lipomedix Pharmaceuticals, Jerusalem, Israel;4. Shaare Zedek Medical Center, Jerusalem, Israel;1. Center for Interventional Endoscopy, Florida Hospital, Orlando, Florida, USA;2. Department of Pathology, Florida Hospital, Orlando, Florida, USA;3. Division of Gastroenterology and Hepatology, Indiana University, Indianapolis, Indiana, USA;4. Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, USA;1. Faculty of Natural Sciences and Mathematics, Institute of Biology, “Ss. Cyril and Methodius” University, P.O. Box 162, 1000 Skopje, Macedonia;2. Faculty of Veterinary Medicine, “Ss. Cyril and Methodius” University, 1000 Skopje, Macedonia |
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| Abstract: | It used to be believed that the use of rat peripheral blood for the micronucleus assay would be difficult because micronucleated erythrocytes are captured and destroyed by the spleen quickly. We have applied an acridine orange (AO) supravital staining method to rat peripheral blood using AO-coated glass slides. Normal and splenectomized SD rats were treated once with mitomycin C (i.p.) or cyclophosphamide (p.o.), and 5 μl of blood was collected at intervals from the tail vein between 0 and 72 h after treatment. For comparison, bone marrow cells were smeared conventionally 30 h after treatment. Although the frequencies of spontaneous and chemically induced micronucleated reticulocytes (MNRETs) from normal rats were lower on average in the highest dose group than those of splenectomized rats, the incidence of micronuclei among type I and II reticulocytes in normal rats at 48 h was almost identical to the incidence of RNA-containing erythrocytes with micronucleus in bone marrow. Thus, we suggest that rat peripheral reticulocytes can be used as target cells for the micronucleus assay. |
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