Tmem100-BAC-EGFP mice to selectively mark and purify embryonic endothelial cells of large caliber arteries in mid-gestational vascular formation
Tmem100-BAC-EGFP mice to selectively mark and purify embryonic endothelial cells of large caliber arteries in mid-gestational vascular formation
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DOI：
10.1002/dvg.23416
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发表时间：
2021-03-02
期刊：
影响因子：
1.5
通讯作者：
Nakagawa, Osamu
中科院分区：
文献类型：
--
作者：
Kinugasa-Katayama, Yumi;Watanabe, Yusuke;Nakagawa, Osamu
胚胎血管的发育是通过不同细胞类型的分化、增殖、迁移和相互作用的复杂排列来实现的，而独特细胞群体的可视化和纯化是利用体内实验模型研究其详细机制的基础。我们先前证明了Tem100是一种新的内皮基因，编码一种小的跨膜蛋白，而Tem100基因缺失的小鼠由于严重的血管形成障碍而表现出胚胎死亡。在本研究中，我们使用包含小鼠Tem100基因的216kb基因组区域构建了一个绿色荧光蛋白报告小鼠品系。一个命名为Tem100-BAC-EGFP小鼠的新品系精确地概括了Tem100在妊娠中期的表达谱，该表达谱在小鼠胚胎的大口径动脉内皮细胞中高度丰富。FACS实验表明，TMem100-BAC-EGFP小鼠能够选择性地纯化特定数量的动脉内皮细胞，这表明它们不仅适用于Tme100表达和功能的研究，也适用于胚胎血管发育中多个内皮细胞亚群的比较分析。
Embryonic vascular development is achieved through the complex arrays of differentiation, proliferation, migration and mutual interaction of different cell types, and visualization as well as purification of unique cell populations are fundamental in studying its detailed mechanisms using in vivo experimental models. We previously demonstrated that Tmem100 was a novel endothelial gene encoding a small transmembrane protein, and that Tmem100 null mice showed embryonic lethality due to severe impairment of vascular formation. In the present study, we generated an EGFP reporter mouse line using a 216 kb genomic region containing mouse Tmem100 gene. A novel line designated as Tmem100-BAC-EGFP mice precisely recapitulated the Tmem100 expression profile at the mid-gestational stage, which was highly enriched in endothelial cells of large caliber arteries in mouse embryos. FACS experiments demonstrated that Tmem100-BAC-EGFP mice served to selectively purify a specific population of arterial endothelial cells, indicating their usefulness not only for the research concerning Tmem100 expression and function but also for comparative analysis of multiple endothelial cell subgroups in embryonic vascular development.