Detection of chromosome malsegregation to the daughter nuclei in cytokinesis-blocked transgenic mouse splenocytes.

Most of the recently developed tests for detecting aneugenic activity of chemicals are based on the induction of micronuclei (MN) in cytokinesis-blocked (CB) binucleated cells. In such a test, aneugens can be discriminated from clastogens by checking for the presence of centromeres in the MN, indicating the loss of whole chromosomes. Tracing particular chromosomes in interphase nuclei using fluorescence in situ hybridization (FISH) with chromosome-specific DNA probes is another method used for detecting numerical chromosome aberrations. Here, we describe a method using a cytokinesis-blocked MN assay in combination with identifying specific chromosomes of mice. For this purpose transgenic mice with foreign DNA inserted in three pairs of their chromosomes were generated. Splenocytes of these mice were cultured and treated in vitro with vinblastine (VBL) or X-rays, followed by recovery in medium containing cytochalasin B. By tracing the marker chromosomes in binucleated splenocytes, reciprocal products of chromosome malsegregation to the daughter nuclei could be easily traced. The results showed that besides clastogenic activity, X-rays also exhibited aneugenic activity. Treatment with vinblastine showed a close relationship between micronuclei induction and chromosome malsegregation, although at higher doses malsegregation processes became more prominent. Simultaneous malsegregation of more than one chromosome was observed frequently, but the three marker chromosomes were found to be randomly involved in this process.