Maternal exposure to the environmental toxicant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induces a variety of defects in compaction-stage embryos, including monopolar spindle formation, errors in chromosome segregation, and fragmentation resulting from aberrant cytokinesis. In this study, we investigated the possibility that a failure in centrosome duplication, separation, or positioning within blastomeres might underlie the observed effects of TCDD on early embryos. The subcellular localization of the centrosomal marker TUBG1 was analyzed in preimplantation embryos collected from female rats exposed to either chronic (50 ng kg⁻¹ wk⁻¹ for 3 wk) or acute (50 ng/kg or 1 μg/kg at proestrus) doses of TCDD. In treated embryos, interphase TUBG1 foci were more abundant and cortically displaced when compared to those in controls. At prophase, some blastomeres exhibited a single large perinuclear TUBG1 aggregate, suggesting a failure in centrosome duplication or separation. Furthermore, the presence of monopolar spindles at metaphase was confirmed by the localization of TUBG1 to the single spindle pole. Therefore, the misregulation of centrosome number and localization, as indicated by TUBG1 staining, may contribute to errors in chromosome segregation and cytokinesis in embryos following maternal TCDD exposure.