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An automated method for discriminating aneugen‐ vs. clastogen‐induced micronuclei
Author(s) -
Torous Dorothea K.,
Dertinger Stephen D.,
Hall Nikki E.,
Tometsko Carol R.
Publication year - 1998
Publication title -
environmental and molecular mutagenesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1
H-Index - 87
eISSN - 1098-2280
pISSN - 0893-6692
DOI - 10.1002/(sici)1098-2280(1998)31:4<340::aid-em6>3.0.co;2-m
Subject(s) - clastogen , micronucleus test , propidium iodide , reticulocyte , mutagen , microbiology and biotechnology , micronucleus , methyl methanesulfonate , flow cytometry , biology , genotoxicity , chemistry , genetics , dna , toxicity , dna damage , apoptosis , organic chemistry , programmed cell death , messenger rna , gene
A flow cytometric (FCM) procedure for quantitating micronucleated reticulocytes in mouse peripheral blood samples was evaluated for its ability to discriminate between aneugen‐ and clastogen‐induced micronuclei (MN). In this experiment, BALB/c mice were injected with 0.9% saline, the model clastogen methyl methanesulfonate (100 mg/kg bw) or the aneugen vincristine (0.2 mg/kg bw). Peripheral blood samples were collected 48 hr after injection and were subsequently fixed and stained for flow cytometric analysis. The staining method utilized FITC‐conjugated anti‐CD71 to differentially label reticulocytes, and the nucleic acid dye propidium iodide to resolve erythrocyte populations with and without micronuclei. The frequency of micronucleated reticulocytes was determined by analyzing 10,000 total reticulocytes per blood sample. A second analysis was performed on each sample whereby the propidium iodide associated fluorescent signals of 250 MN were collected and graphed as a single‐parameter histogram. The histogram statistic “median channel” was recorded for each sample and provided a quantitative description of MN distribution according to DNA content. Cumulatively, the results of this study suggest that 1) flow cytometry can be employed to measure the incidence of MN resulting from clastogenic or aneugenic activity, and 2) MN resulting from aneugenscan be discriminated from those arising spontaneously or from clastogen treatment based on flow cytometric analysis of DNA content. Environ. Mol. Mutagen. 31:340–344, 1998. © 1998 Wiley‐Liss, Inc.