Institute of Physiology I, University of Bonn, Sigmund-Freud-Str. 25, 53127, Bonn, Germany.
Basic Res Cardiol. 2012 Mar;107(2):257. doi: 10.1007/s00395-012-0257-5. Epub 2012 Mar 1.
Vessel formation is of critical importance for organ function in the normal and diseased state. In particular, the labeling and quantitation of small vessels prove to be technically challenging using current approaches. We have, therefore, established a transgenic embryonic stem (ES) cell line and a transgenic mouse model where the vascular endothelial growth factor receptor VEGFR-1 (flt-1) promoter drives the expression of the live reporter eGFP. Fluorescence microscopy and immunostainings revealed endothelial-specific eGFP labeling of vascular networks. The expression pattern recapitulates that of the endogenous flt-1 gene, because small and large vessels are labeled by eGFP during embryonic development; after birth, the expression becomes more restricted to small vessels. We have explored this in the cardiovascular system more in detail and found that all small vessels and capillaries within the heart are strongly eGFP+. In addition, myocardial injuries have been induced in transgenic mice and prominent vascular remodeling, and an increase in endothelial cell area within the peri-infarct area could be observed underscoring the utility of this mouse model. Thus, the transgenic flt-1/eGFP models are powerful tools to investigate and quantify vascularization in vivo and to probe the effect of different compounds on vessel formation in vitro.
血管生成对于正常和疾病状态下的器官功能至关重要。特别是,使用当前方法对小血管进行标记和定量证明具有技术挑战性。因此,我们建立了一种转基因胚胎干细胞(ES)细胞系和一种转基因小鼠模型,其中血管内皮生长因子受体 VEGFR-1(flt-1)启动子驱动活报告基因 eGFP 的表达。荧光显微镜和免疫染色显示血管网络的内皮特异性 eGFP 标记。表达模式再现了内源性 flt-1 基因的表达模式,因为在胚胎发育过程中小血管和大血管均被 eGFP 标记;出生后,表达变得更局限于小血管。我们在心血管系统中更详细地研究了这一点,并发现心脏内的所有小血管和毛细血管均强烈表达 eGFP+。此外,在转基因小鼠中诱导心肌损伤,并观察到明显的血管重塑和梗塞周围区域内皮细胞面积增加,这突出了该小鼠模型的实用性。因此,转基因 flt-1/eGFP 模型是研究体内血管生成和体外探测不同化合物对血管生成影响的有力工具。