Department of Obstetrics and Gynecology, Akita University Graduate School of Medicine, Akita, Japan.
Department of Obstetrics and Gynecology, Omagari Kousei Medical Center, Akita, Japan.
PLoS One. 2021 Jan 29;16(1):e0246337. doi: 10.1371/journal.pone.0246337. eCollection 2021.
Studies have shown that some electrolytes, including Na+ and K+, play important roles in embryonic development. However, these studies evaluated these electrolytes by using inhibitors or knockout mice, with no mention on the changes in the intracellular electrolyte concentrations during embryogenesis. In this study, we used the electrolyte indicators CoroNa Green AM and ION Potassium Green-2 AM to directly visualise intracellular concentrations of Na+ and K+, respectively, at each embryonic developmental stage in mouse embryos. We directly observed intracellular electrolyte concentrations at the morula, blastocyst, and hatching stages. Our results revealed dynamic changes in intracellular electrolyte concentrations; we found that the intracellular Na+ concentration decreased, while K+ concentration increased during blastocoel formation. The degree of change in intensity in response to ouabain, an inhibitor of Na+/K+ ATPase, was considered to correspond to the degree of Na+/K+ ATPase activity at each developmental stage. Additionally, after the blastocyst stage, trophectoderm cells in direct contact with the blastocoel showed higher K+ concentrations than in direct contact with inner cell mass, indicating that Na+/K+ ATPase activity differs depending on the location in the trophectoderm. This is the first study to use CoroNa Green AM and ION Potassium Green-2 AM in mouse embryos and visualise electrolytes during embryonic development. The changes in electrolyte concentration observed in this study were consistent with the activity of Na+/K+ ATPase reported previously, and it was possible to image more detailed electrolyte behaviour in embryo cells. This method can be used to improve the understanding of cell physiology and is useful for future embryonic development studies.
研究表明,包括 Na+ 和 K+ 在内的一些电解质在胚胎发育中发挥着重要作用。然而,这些研究通过使用抑制剂或敲除小鼠来评估这些电解质,并未提及胚胎发生过程中细胞内电解质浓度的变化。在这项研究中,我们使用电解质指示剂 CoroNa Green AM 和 ION Potassium Green-2 AM,分别直接观察到小鼠胚胎在每个胚胎发育阶段的细胞内 Na+和 K+浓度。我们直接观察了桑葚胚、囊胚和孵化阶段的细胞内电解质浓度。我们的结果显示细胞内电解质浓度发生了动态变化;我们发现,在囊胚腔形成过程中,细胞内 Na+浓度降低,而 K+浓度升高。对哇巴因(Na+/K+ATP 酶抑制剂)的反应强度变化程度被认为与每个发育阶段的 Na+/K+ATP 酶活性相对应。此外,在囊胚阶段之后,与囊胚腔直接接触的滋养外胚层细胞显示出比与内细胞团直接接触更高的 K+浓度,这表明 Na+/K+ATP 酶活性取决于滋养外胚层的位置。这是首次在小鼠胚胎中使用 CoroNa Green AM 和 ION Potassium Green-2 AM 来可视化胚胎发育过程中的电解质,并观察到电解质浓度的变化与之前报道的 Na+/K+ATP 酶活性一致,并且可以更详细地观察胚胎细胞中的电解质行为。该方法可用于提高对细胞生理学的理解,对未来的胚胎发育研究也很有用。