KINETOCHORE IDENTIFICATION IN MICRONUCLEI IN MOUSE BONE-MARROW ERYTHROCYTES - AN ASSAY FOR THE DETECTION OF ANEUPLOIDY-INDUCING AGENTS

An in vivo micronucleus assay using mouse bone marrow for identifying the ability of chemicals to induce aneuploidy and/or chromosome breaks is described. Micronucleus formation in bone-marrow erythrocytes of mice is commonly used as an index for evaluating the clastogenicity of environmental agents. However, micronuclei may also originate from intact lagging chromosomes resulting from the effect of aneuploidy-inducing agents. We have used immunofluorescent staining anti-kinetochore antibodies to classify micronuclei for the presence or absence of kinetochores. Micronuclei positive for kinetochores are assumed to contain intact chromosomes and result from induced aneiploidy; while those negative for kinetochores contain acenteric chromosomal fragments and originate from clastogenic events. The assay was evaluated using X-irradiation response (a known clasge agent). A dose-related response for Micronuclei induced by X-irradiation were negative for kinetochores while the majority of the micronuclei fro resulting from vincristine treatment contained kinetochores. Thus, the micronucleus assay in combination with immunofluorescent staining for kinetochores may provide a useful method to simultaneously assess the ability of chemicals to induce aneuploidy and/or chromosome breaks. © 1990.