Chromosome analysis of mouse one-cell androgenones derived from a sperm nucleus exposed to topoisomerase II inhibitors at pre- and post-fertilization stages
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Mouse spermatozoa and androgenetic one-cell embryos (androgenones) at various developmental stages were exposed to etoposide (1 μM), a topoisomerase II (topo II) poison, or to either of two catalytic inhibitors: ICRF-193 (10 μM) or merbarone (50 μM), for 2 h in order to study the clastogenic effects of these drugs on remodeled sperm chromatin. None of the drugs induced structural chromosome aberrations in condensed chromatin of spermatozoa. However, etoposide and merbarone exerted strong clastogenic actions on remodeled chromatin of androgenones. Expanding chromatin was most sensitive to both of these drugs at the time of pronuclear formation, as nearly 100% of androgenones exposed at this stage displayed structural chromosome aberrations. ICRF-193 did not affect sperm chromatin at all remodeling stages. A majority of the aberrations induced by etoposide and merbarone were of the chromosome-type. Chromosome exchanges, including translocation, dicentric, and ring chromosomes, preferentially appeared following exposure at the early stages of chromatin remodeling. Thus, despite their different modes of topo II inhibition, etoposide and merbarone showed similar clastogenic actions on remodeled sperm chromatin. These results suggest that the formation of transient DNA cleavage, mediated by ooplasmic topo II, accompanies the remodeling. The present findings provide insight into the mechanisms by which structural aberrations are generated in paternal chromosomes.
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