126. IMPLICATIONS FOR FETAL AND PLACENTAL DEVELOPMENT FOLLOWING MITOCHONDRIAL PERTURBATION IN THE EMBRYO

Abstract

The environment an embryo is exposed to can profoundly influence peri- and post-natal development despite having some capacity to adapt. Whilst the mechanisms responsible remain largely unknown, mitochondria are a likely target. In this study we deliberately perturbed mitochondrial function in the mouse embryo, using a model we have established that shows step-wise changes in embryo metabolism and development. The aim of this study was to provide direct evidence implicating mitochondrial dysfunction in the embryo with perturbed fetal and placental development. Zygote stage embryos were recovered from superovulated female mice and cultured in control conditions to the 2-cell stage. Embryos were then allocated to one of three treatments; control media (0μM-AOA), 5μM or 50μM of the known mitochondrial inhibitor, Amino-Oxyacetate, in the absence of pyruvate (5μM-AOA, 50μM-AOA). Embryos were cultured to the blastocyst stage, then transferred to pseudopregnant recipients, with fetal and placental parameters measured on day 18 of pregnancy. Implantation rates and fetal survival for both 5μM-AOA and 50μM-AOA was comparable to control embryos. For 5μM-AOA there was a significant reduction in placental weight (P=0.02) but normal fetal weight, and a significant increase in fetal: placental weight ratio (P=0.002) relative to the control, suggesting increased placental efficiency. When mitochondria were further perturbed (50μM-AOA), the fetuses and placentas were both considerably compromised: that is, decreased fetal and placental weights (P=0.002), reduced placental diameter (P=0.03) and decreased fetal crown rump length (P=0.07). This study demonstrates that mitochondrial function in the embryo impacts on peri-natal development, providing compelling evidence for mitochondrial function involvement in the mechanisms underpinning “embryo programming”. This data suggests a threshold effect, whereby embryos can only adapt up until a point after which development is compromised. Further elucidating these mechanisms is important for understanding how maternal environments and embryo culture systems determine development of future offspring.