Correlation of the Mitochondrial Activity of Two‐Cell Embryos Produced In Vitro and the Two‐Cell Block In Kunming and B6C3F1 Mice

The correlation between the early embryonic block to development and mitochondrial activity was investigated comparing two‐cell embryos produced in vitro from Kunming (KM) and B6C3F1 mice. One‐cell embryos were obtained from two species of hybrids (female KM mice mated with KM males and female B6C3F1 mice mated with KM males) and cultured for 84 hr in M16 media. The mitochondrial membrane potential, ATP content, and reactive oxygen species levels were measured in the resulting KM and B6C3F1 two‐cell embryos. Mitochondrial membrane potential and ATP content were also determined in KM and B6C3F1 metaphase II eggs. The results showed that the two‐cell block was observed in cultured KM embryos but not in B6C3F1 embryos. Mitochondrial membrane potential and ATP content of KM two‐cell embryos were significantly lower than in B6C3F1 two‐cell embryos (P < 0.01). Interestingly, the reactive oxygen species levels of KM two‐cell embryos were significantly lower than their B6C3F1 counterparts (P < 0.01). There was no difference in mitochondrial membrane potential and ATP content between KM and B6C3F1 metaphase II eggs. It is concluded that KM mice have an early two‐cell embryo block and that a possible “blocking” mechanism is the lower mitochondrial membrane potential and ATP content in these embryos. The results suggest a new approach for overcoming early embryonic development block, that of manipulating mitochondrial activity. Anat Rec, 292:661–669, 2009. © 2009 Wiley‐Liss, Inc.