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syndecan-1的表达在小鼠F9畸胎癌细胞向内胚层和脏壁内胚层分化过程中发生变化。

Expression of syndecan-1 changes during the differentiation of visceral and parietal endoderm from murine F9 teratocarcinoma cells.

作者信息

Jiang R, Kato M, Bernfield M, Grabel L B

机构信息

Department of Biology, Wesleyan University, Middletown, CT 06459, USA.

出版信息

Differentiation. 1995 Nov;59(4):225-33. doi: 10.1046/j.1432-0436.1995.5940225.x.

Abstract

F9 teratocarcinoma stem cells treated with retinoic acid differentiate in suspension into embryoid bodies with an outer layer of visceral endoderm surrounding a core of largely undifferentiated cells. The visceral endoderm-containing embryoid bodies, when plated onto an extracellular matrix coating, give rise to parietal endoderm outgrowth. These in vitro cell cultures mimic both geometrically and biochemically the differentiation of visceral and parietal endoderm in the early mouse embryo and, thus, were used as a model system for the study of molecular and cellular mechanisms underlying the differentiation of the extraembryonic endoderm lineages. We have investigated the expression of syndecan-1, an integral membrane proteoglycan that binds to multiple components of the extracellular matrix and basic FGF, during visceral endoderm differentiation and parietal endoderm outgrowth. Syndecan-1 immunostaining is detected on all cell surfaces in the undifferentiated embryoid bodies and in the differentiating embryoid bodies prior to the formation of the visceral endoderm. Following the differentiation of visceral endoderm, syndecan-1 localizes predominantly to the basal surface of this epithelial layer, while syndecan-1 staining in the core of differentiated embryoid bodies is faint. Quantitation of cell associated syndecan-1 indicates that syndecan-1 is down-regulated during embryoid body differentiation. However, northern analysis shows that the amounts of steady-state syndecan-1 mRNA are the same in undifferentiated versus differentiated embryoid bodies, suggesting post-transcriptional regulation of syndecan-1 expression in the differentiating embryoid body. Analysis of syndecan-1 distribution in the outgrowth culture by immunofluorescence demonstrates that syndecan-1 is absent from the cell surface of parietal endoderm. However, a substantial amount of syndecan-1 is detected inside parietal endoderm cells. While all three cell types release syndecan-1 ectodomain into the culture medium, the parietal endoderm outgrowth releases more syndecan-1 ectodomain than the differentiated embryoid body. These data suggest that the post-transcriptional control and post-translational shedding of syndecan-1 from the cell surface are developmentally regulated during the differentiation of visceral to parietal endoderm and the migration of parietal endoderm.

摘要

用视黄酸处理的F9畸胎瘤干细胞在悬浮培养中分化为胚状体,其外层为脏内胚层,围绕着一个主要由未分化细胞组成的核心。当将含有脏内胚层的胚状体接种到细胞外基质涂层上时,会产生壁内胚层生长物。这些体外细胞培养物在几何形状和生化方面都模拟了小鼠早期胚胎中脏内胚层和壁内胚层的分化,因此被用作研究胚外内胚层谱系分化的分子和细胞机制的模型系统。我们研究了syndecan-1的表达,syndecan-1是一种整合膜蛋白聚糖,可与细胞外基质的多种成分和碱性成纤维细胞生长因子结合,研究其在脏内胚层分化和壁内胚层生长过程中的表达情况。在未分化的胚状体以及形成脏内胚层之前的分化胚状体的所有细胞表面均检测到syndecan-1免疫染色。脏内胚层分化后,syndecan-1主要定位于该上皮层的基底表面,而在分化胚状体核心中的syndecan-1染色较弱。对细胞相关syndecan-1的定量分析表明,syndecan-1在胚状体分化过程中被下调。然而,Northern分析显示,未分化与分化的胚状体中稳态syndecan-1 mRNA的量相同,这表明在分化的胚状体中syndecan-1的表达存在转录后调控。通过免疫荧光分析syndecan-1在生长培养物中的分布表明,壁内胚层的细胞表面不存在syndecan-1。然而,在壁内胚层细胞内部检测到大量的syndecan-1。虽然所有三种细胞类型都将syndecan-1胞外域释放到培养基中,但壁内胚层生长物释放的syndecan-1胞外域比分化的胚状体更多。这些数据表明,在从脏内胚层到壁内胚层的分化以及壁内胚层的迁移过程中,syndecan-1从细胞表面的转录后调控和翻译后脱落受到发育调控。

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