Differential Expression of Ezrin and CLP36 in the Two Layers of Syncytiotrophoblast in Rats
Differential Expression of Ezrin and CLP36 in the Two Layers of Syncytiotrophoblast in Rats
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DOI：
10.1248/bpb.33.1400
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发表时间：
2010-08-01
影响因子：
2
通讯作者：
Nakashima, Emi
中科院分区：
文献类型：
--
作者：
Higuchi, Kei;Iizasa, Hisashi;Nakashima, Emi
合体滋养层细胞在人类中由单层组成，在啮齿动物中由两层组成，即SynI和SynII。合体滋养层负责调节母体对药物的转运。转运蛋白在合体滋养层细胞顶端和基底膜上的极性分布在底物的载体运输方面对胎盘功能是重要的，但控制合体滋养层细胞蛋白质分布的机制尚不清楚。我们已经建立了大鼠合体滋养层细胞系，tr-TBT 18d-1和tr-TBT 18d-2，分别保留了SynI和SynII的特征。在这项研究中，我们的目的是通过使用这些细胞系来表征这两层细胞的基因表达谱。DNA芯片分析表明，包括细胞骨架结合蛋白Ezrin和CLP36在内的超过25mRNAs在tr-TBT 18d-1和tr-TBT 18d-2之间存在差异表达。实时定量聚合酶链式反应(RT-PCR)分析表明，与相应的对照相比，RT-TBT 18d-1的Ezrin、CLP36、CCN1和CCN2的mRNA表达水平较高，而ELF-1a、hsc70和Flot2的mRNA的表达水平较高。免疫组织化学分析表明，Ezrin在大鼠胎盘绒毛中有表达，定位于tr-TBT 18d-1的顶膜，而CLP36位于tr-TBT 18d-1的顶侧和基底侧。Ezrin在妊娠14天和18天表达最高，并且在Ezrin/Radisin/moesin(ERM)家族成员中表达最高。这些结果可能有助于阐明合体滋养层细胞极化的控制机制以及双层上皮层在大鼠和小鼠中的意义。
The syncytiotrophoblast, which regulates maternal fetal transfer of drugs, consists of a single layer in humans, but two layers, i.e., SynI and SynII, in rodents. Polar distribution of transporters in the apical and basal plasma membranes of syncytiotrophoblast is important for placental function in terms of vectorial transport of substrates, but the mechanisms that control protein distribution in the syncytiotrophoblast remain unclear. We have previously established rat syncytiotrophoblast cell lines, TR-TBT 18d-1 and TR-TBT 18d-2, which retain characteristics of SynI and SynII, respectively. In this study, we aimed to characterize the gene expression profiles in the two layers by using these cell lines. DNA microarray analysis indicated that more than 25 mRNAs, including cytoskeleton binding proteins, ezrin and CLP36, are differentially expressed between TR-TBT 18d-1 and TR-TBT 18d-2. Quantitative real time-polymerase chain reaction (PCR) analysis indicated that mRNA expression of ezrin, CLP36, CCN1, and CCN2 is higher in TR-TBT 18d-1 and mRNA expression of elf-la, hsc70 and flot2 is higher in TR-TBT 18d-2, compared with their counterparts. Immunohistochemical analysis indicated that ezrin is expressed in rat placental villi in vivo, and is located on the apical membranes of TR-TBT 18d-1, while CLP36 is located in the apical and basal sides of TR-TBT 18d-1. The expression of ezrin was highest at gestational days 14 and 18 and was highest among the ezrin/radixin/moesin (ERM) family members. These results may help to clarify the mechanisms controlling polarization of the syncytiotrophoblast and the significance of the double epithelial layers in rat and mouse.