Intracellular aH of the preimplantation mouse embryo: effects of extracellular aH and weak acids

摘要

Although intracellular aH (pHi), is a regulator of numerous biological processes, it has received relatively little attention with regard to the physiology of the mammalian preimplantation embryo. Interestingly, there is some controversy as to whether the early embryo can recover from an acid load. The significance of this is that two constituents of mouse embryo culture media are pyruvate and lactate. These carboxylic acids are utilised by the early mouse embryo for energy production. However, as weak acids, pyruvate and lactate may induce perturbations in the phi and thus alter the physiology of the embryo. The aims of this study were therefore to measure the phi of the mouse preimplantation embryo and to determine the effect of lactate on phi at different developmental stages. The phi was measured using the ratio-metric fluorophore carboxy-seminaphthorhodafluor-l-acetoxymethylester (SNARF-1)in conjunction with confocal microscopy. The phi increased significantly with development from the zygote to the morula stage. Furthermore, at concentrations greater than 5 mM, lactate caused the phi of the zygote to become significantly more acidic. It was demonstrated that facilitative transport in association with a smaller passive component was responsible for the movement of lactate into the zygote. Metabolic studies revealed that, through their acidifying effect, weak acids caused a reduction in glycolytic activity in the early embryo. In contrast, the phi of the compacted embryo remained unchanged by the presence of lactate in the external media. Furthermore, incubation with weak acids did not affect the rate of glycolysis in the morula. These data suggest that, by the generation of a transporting epithelium at compaction, the embryo develops the ability to regulate phi against an acid load.