Kinetochore localization in micronucleated cytokinesis-blocked Chinese hamster ovary cells: a new and rapid assay for identifying aneuploidy-inducing agents.
We have developed a modified micronucleus assay using an antikinetochore antibody and cytokinesis-blocked Chinese hamster ovary cells as a simple and rapid method for detecting aneuploidy-inducing agents. The presence of a kinetochore in a micronucleus of a binucleated cell indicates a cell with a high probability for aneuploidy following cytokinesis. The method requires minimal training to perform and score and can readily distinguish aneuploidy-inducing agents from clastogens. Micronucleated cells treated with the aneuploidy-inducing agents benomyl and vinblastine sulfate contained a kinetochore-positive micronucleus 92% and 94% of the time whereas micronucleated cells treated with the clastogen methyl methanesulfonate contained a kinetochore-positive micronucleus only 11% of the time. This relatively simple method for distinguishing aneuploidy-inducing agents from clastogenic agents may be used as a routine genotoxicity assay to identify environmental and therapeutic agents with aneuploidy-inducing properties.
Eastmond
D A
DA
Biomedical Sciences Division, Lawrence Livermore National Laboratory, CA 94550.
Tucker
J D
JD
D000782:Aneuploidy; D000818:Animals; D001542:Benomyl / Q000633:toxicity; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002503:Centromere / Q000648:ultrastructure*; D002875:Chromosomes / Q000648:ultrastructure*; D006224:Cricetinae; D003412:Cricetulus; D004305:Dose-Response Relationship, Drug; D005260:Female; D005455:Fluorescent Antibody Technique; D008741:Methyl Methanesulfonate / Q000633:toxicity; D015162:Micronucleus Tests / Q000379:methods*; D010053:Ovary; D014747:Vinblastine / Q000633:toxicity
D014747:Vinblastine; D008741:Methyl Methanesulfonate; D001542:Benomyl