Department of Molecular Physiology, National Cerebral and Cardiovascular Center Research Institute, Osaka, Japan.
Graduate School of Medical Sciences, Nara Medical University, Kashihara, Nara, Japan.
Genesis. 2021 Apr;59(4):e23416. doi: 10.1002/dvg.23416. Epub 2021 Mar 2.
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.
胚胎血管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,使用体内实验模型研究其详细机制时,对独特细胞群体进行可视化和纯化是非常重要的。我们之前证明,Tmem100 是一种新的内皮基因,编码一种小的跨膜蛋白,Tmem100 缺失小鼠由于血管形成严重受损而导致胚胎致死。在本研究中,我们使用包含小鼠 Tmem100 基因的 216kb 基因组区域生成了一个 EGFP 报告鼠系。一个新的命名为 Tmem100-BAC-EGFP 的小鼠系精确地重现了 Tmem100 在中期妊娠阶段的表达模式,该模式在小鼠胚胎的大口径动脉内皮细胞中高度富集。FACS 实验表明,Tmem100-BAC-EGFP 小鼠可用于选择性纯化特定的动脉内皮细胞群体,这表明它们不仅可用于研究 Tmem100 的表达和功能,还可用于胚胎血管发育中多个内皮细胞亚群的比较分析。