VEGF-induced mobilization of caveolae and increase in permeability of endothelial cells
VEGF-induced mobilization of caveolae and increase in permeability of endothelial cells
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DOI：
10.1152/ajpcell.00292.2001
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发表时间：
2002-05-01
影响因子：
5.5
通讯作者：
Goligorsky, MS
中科院分区：
文献类型：
--
作者：
Chen, J;Braet, F;Goligorsky, MS
肾小球上皮细胞（GEC）是已知的血管内皮生长因子（VEGF）产生部位。我们建立了永生化的大鼠GEC，它保留了产生VEGF的能力。GEC表达的亚型定义为VEGF-205、-188、-120和-164。内皮细胞的电阻培养的GEC条件下的矩阵，一个指标的渗透性单层溶质，显着增加了治疗与VEGF的中和多克隆抗体和VEGF-165下降。用绿色荧光蛋白-小窝蛋白构建体转染内皮细胞，活体共聚焦显微镜显示VEGF导致用小窝蛋白装饰的跨细胞细长结构的快速出现。内皮细胞的透射电子显微镜显示，小窝进行快速内化和融合后30分钟的应用VEGF-165。后来（36小时），血管内皮生长因子预处理的内皮细胞开发窗孔，并显示电阻下降。肾小球免疫电镜证实VEGF定位于足细胞和基底膜。总之，永生化GEC保留了合成VEGF的能力。基质沉积和可溶性VEGF导致小窝表达的增强、它们的分裂和融合、细长小窝蛋白修饰结构的形成以及窗孔的最终形成，这两者都是内皮渗透性增加的原因。
Glomerular epithelial cells (GEC) are a known site of vascular endothelial growth factor (VEGF) production. We established immortalized rat GEC, which retained the ability to produce VEGF. The isoforms expressed by GEC were defined as VEGF-205, -188, -120, and -164. The electrical resistance of endothelial cells cultured on GEC-conditioned matrix, an indicator of the permeability of monolayers to solutes, was significantly increased by the treatment with the neutralizing polyclonal antibodies to VEGF and decreased by VEGF-165. Transfection of endothelial cells with green fluorescence protein-caveolin construct and intravital confocal microscopy showed that VEGF results in a rapid appearance of transcellular elongated structures decorated with caveolin. Transmission electron microscopy of endothelial cells showed that caveolae undergo rapid internalization and fusion 30 min after application of VEGF-165. Later (36 h), endothelial cells pretreated with VEGF developed fenestrae and showed a decrease in electrical resistance. Immunoelectron microscopy of glomeruli confirmed VEGF localization to podocytes and in the basement membrane. In summary, immortalized GEC retain the ability to synthesize VEGF. Matrix-deposited and soluble VEGF leads to the enhancement of caveolae expression, their fission and fusion, formation of elongated caveolin-decorated structures, and eventual formation of fenestrae, both responsible for the increase in endothelial permeability.