Na+/K+ ATPase α1 and β3 subunits are localized to the basolateral membrane of trophectoderm cells in human blastocysts.
Na+/K+ ATPase α1 and β3 subunits are localized to the basolateral membrane of trophectoderm cells in human blastocysts.
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Na ~+/K ~+ ATP酶α1和β3亚基定位于人胚泡滋养外胚层细胞的基底外侧膜。
DOI：
10.1093/humrep/deac124
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发表时间：
2022-06-30
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Na+/K+ ATP酶在人胚泡中的特异性表达和定位与胚胎的发育行为之间是否存在关系？Na+/K+ ATP酶α1、β1和β3是人胚泡中表达的主要亚型，其mRNA表达水平与胚泡扩张率之间无相关性。在小鼠胚胎中，Na+/K+ ATP酶α1和β1在滋养外胚层（TE）细胞的基底外侧膜中表达，并被认为参与囊胚腔形成（空化）。研究人员分析了2009年至2018年期间在一所大学医院接受IVF和胚胎移植的11名患者的20个多余胚胎。冷冻和解冻后第5天的人囊胚，使用延时成像观察其发育行为24 h，并使用定量RT-PCR（RT-qPCR）检测Na+/K+ ATP酶亚型的表达。然后使用荧光免疫染色将表达的同种型定位在胚泡中。RT-qPCR结果显示Na+/K+ ATP酶α1、β1和β3亚型在人囊胚中表达。亚型α1和β3定位于TE细胞的基底外侧膜，β1定位于TE细胞之间。β3 mRNA的高表达与孵化率相关（P = 0.0261）。N/A. mRNA的表达和感兴趣的蛋白质的定位得到了验证，但我们还不能进行功能分析。在各种Na+/K+ ATP酶亚型中，α1、β1和β3 mRNA的表达水平明显高于人胚泡中的其他亚型。由于α1和β3通过荧光免疫染色定位于基底外侧膜，我们认为这些亚基有助于囊胚腔的扩张。β1亚型位于TE细胞之间，可能参与紧密连接的形成，如先前在小鼠胚胎中所报道的。这项工作得到了日本科学研究所KAKENHI（https：//www.jsps.go.jp/english/index.html）的支持，资助号为17 K11215。资助者在研究设计、数据收集和分析、出版决定或手稿编写中没有任何作用。作者没有利益冲突。
Is there a relation between specific Na+/K+ ATPase isoform expression and localization in human blastocysts and the developmental behavior of the embryo? Na+/K+ ATPase α1, β1 and β3 are the main isoforms expressed in human blastocysts and no association was found between the expression level of their respective mRNAs and the rate of blastocyst expansion. In mouse embryos, Na+/K+ ATPase α1 and β1 are expressed in the basolateral membrane of trophectoderm (TE) cells and are believed to be involved in blastocoel formation (cavitation). A total of 20 surplus embryos from 11 patients who underwent IVF and embryo transfer at a university hospital between 2009 and 2018 were analyzed. After freezing and thawing Day 5 human blastocysts, their developmental behavior was observed for 24 h using time-lapse imaging, and the expression of Na+/K+ ATPase isoforms was examined using quantitative RT-PCR (RT-qPCR). The expressed isoforms were then localized in blastocysts using fluorescent immunostaining. RT-qPCR results demonstrated the expression of Na+/K+ ATPase α1, β1 and β3 isoforms in human blastocysts. Isoforms α1 and β3 were localized to the basolateral membrane of TE cells, and β1 was localized between TE cells. A high level of β3 mRNA expression correlated with easier hatching (P = 0.0261). N/A. The expression of mRNA and the localization of proteins of interest were verified, but we have not been able to perform functional analysis. Of the various Na+/K+ ATPase isoforms, expression levels of the α1, β1 and β3 mRNAs were clearly higher than other isoforms in human blastocysts. Since α1 and β3 were localized to the basolateral membrane via fluorescent immunostaining, we believe that these subunits contribute to the dilation of the blastocoel. The β1 isoform is localized between TE cells and may be involved in tight junction formation, as previously reported in mouse embryos. This work was supported by the JSPS KAKENHI (https://www.jsps.go.jp/english/index.html), grant number 17K11215. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors have no conflicts of interest.
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