Chemical induction of quadruple and octuple chromosomes in Chinese hamster CHO-K1 cells and relationship between their three-dimensional structure and spatial distribution of BrdU-labeled chromatids

Abstract

Double endoreduplication of Chinese hamster CHO-K1 cells that exhibited quadruple chromosomes at metaphase was induced by a combination of rotenone and ammonium vanadate treatments. Analysis of sister chromatid differential staining patterns (using 5-bromo-2'-deoxyuridine) revealed that approximately 50% of the quadruple chromosomes did not keep the scheme of "outside replication" of DNA. Based on the ratio of the staining patterns observed, we suggest that the two diplochromosomes forming a quadruple chromosome are held together by a physical link connecting the two original chromatids. Metaphases with octuple chromosomes were also produced by the same treatment. Each chromosome constituting an octuple chromosome was longer and thinner than ordinary metaphase chromosomes. This suggests incomplete chromosome condensation at metaphase. The majority of octuple chromosomes showed the eight constituent chromosomes to be so enmeshed that a planar alignment could not be observed in air-dried preparations.