Rehn Kira, Wong Kuan Shen, Balciunas Darius, Sumanas Saulius
Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.
Int J Dev Biol. 2011;55(6):613-8. doi: 10.1387/ijdb.103249kp.
Aquaporin 1 (Aqp1) is a water channel protein, expressed widely in microvascular endothelial cells and implicated in mammalian tumor angiogenesis. However, its developmental expression has not yet been characterized in great detail. An enhancer trap screen was performed using a Tol2-derived GFP reporter in zebrafish embryos. An insertional Et(GBT-B1)tpl1 line was identified that has reporter insertion in the vicinity of the aqp1a gene. We further characterized the embryonic expression pattern of this GFP reporter line, as well as that of endogenous aqp1a. Both endogenous aqp1a and reporter GFP expression were restricted to the vascular endothelial cells within the dorsal aorta, cranial, intersegmental and other secondary vessels, but were absent in the axial venous vasculature. In addition, endogenous aqp1a expression was observed in both primitive and definitive hematopoietic erythroid progenitors, as well as in the otic vesicle, swim bladder, pneumatic duct, intestine and a subset of neurons within the retina and the midbrain-hindbrain region. We further show that gata1 and etsrp/etv2 function is required for hematopoietic and endothelial aqp1a expression, respectively. Aqp1a expression is restricted to endothelial and erythroid cells during early embryogenesis. The transgenic Et(GBT-B1)tpl1 line recapitulates endogenous endothelial aqp1a expression. Because currently very few reporter lines can differentiate between arterial and venous endothelial cells, the Et(GBT-B1)tpl1 transgenic line and characterization of the aqp1a expression pattern will be useful for future studies of endothelial and arterial-venous differentiation.
水通道蛋白1(Aqp1)是一种水通道蛋白,在微血管内皮细胞中广泛表达,并与哺乳动物肿瘤血管生成有关。然而,其在发育过程中的表达尚未得到详细表征。在斑马鱼胚胎中使用源自Tol2的绿色荧光蛋白(GFP)报告基因进行了增强子陷阱筛选。鉴定出一个插入性的Et(GBT-B1)tpl1品系,其报告基因插入到aqp1a基因附近。我们进一步表征了该GFP报告基因品系以及内源性aqp1a的胚胎表达模式。内源性aqp1a和报告基因GFP的表达均局限于背主动脉、颅部、节间和其他二级血管内的血管内皮细胞,但在轴向静脉血管系统中不存在。此外,在内源性原始和定型造血红系祖细胞以及耳泡、鳔、气动导管、肠道以及视网膜和中脑-后脑区域的一部分神经元中观察到了内源性aqp1a的表达。我们进一步表明,gata1和etsrp/etv2的功能分别是造血和内皮细胞中aqp1a表达所必需的。在早期胚胎发育过程中,Aqp1a的表达局限于内皮细胞和红系细胞。转基因Et(GBT-B1)tpl1品系概括了内源性内皮细胞aqp1a的表达。由于目前很少有报告基因品系能够区分动脉和静脉内皮细胞,因此Et(GBT-B1)tpl1转基因品系以及aqp1a表达模式的表征将有助于未来内皮细胞和动静脉分化的研究。