2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a potent teratogen in several animal species, especially during the period of organogenesis. The purpose of this study was to test the hypothesis that TCDD has direct effects on the earliest stages of murine embryonic development, namely the preimplantation stages. Three endpoints were measured: 1) embryo cell number, a measure of embryo viability, 2) competitive embryonic cell proliferation utilizing chimeric embryos, another measure of embryo viability, and 3) cavitation rate, a functional measure of trophectoderm differentiation. Neither embryo cell numbers nor competitive embryonic cell proliferation (chimera assay) were affected by TCDD, either when the mother was dosed in vivo (prior to fertilization), or when 2-cell embryos were dosed in vitro. However, cavitation rates of in vitro-dosed embryos, in the presence of 10% fetal bovine serum, were significantly higher for TCDD than controls, suggesting that TCDD accelerated differentiation of murine preimplantation embryos. Taken together, these results demonstrate that: 1) TCDD can act directly on the murine preimplantation embryo, and 2) TCDD's actions are primarily on accelerated differentiation and not on embryo viability. To our knowledge, this is the earliest stage of mammalian development during which TCDD has been shown to exert an effect.