Inducible expression of vascular endothelial growth factor in adult mice causes severe proliferative retinopathy and retinal detachment
Inducible expression of vascular endothelial growth factor in adult mice causes severe proliferative retinopathy and retinal detachment
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DOI：
10.1016/s0002-9440(10)64891-2
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发表时间：
2002-02-01
影响因子：
6
通讯作者：
Campochiaro, PA
中科院分区：
文献类型：
--
作者：
Ohno-Matsui, K;Hirose, A;Campochiaro, PA
转基因的。具有由视紫红质启动子驱动的血管内皮生长因子（VEGF）的小鼠（rho/VEGF小鼠）产生新血管形成，其起源于视网膜的深毛细血管床并生长到视网膜下空间。在rho/VEGF小鼠中，光感受器中的VEGF表达开始于出生后第5天至第7天，此时深毛细血管床正在发育。一个重要的问题是深毛细血管床的发育阶段是否是新生血管发生的关键。此外，尽管rho/VEGF小鼠对于眼部新生血管形成的研究非常有用，但是对于某些应用，VEGF表达的早期发作是不利的。在这项研究中，我们使用的反向四环素反式激活因子（rtTA）诱导型启动子系统耦合到视紫红质或interphotoreceptor类维生素A结合蛋白（IRBP）启动子来控制VEGF转基因表达的光感受器的发病时间。在不存在强力霉素的情况下，成年双转基因rho/rtTA-TRE/VEGF或IRBP/rtTA-TRE/VEGF小鼠显示很少的VEGF转基因表达并且没有表型。在饮用水中添加强力霉素导致显著的转基因表达和3至4天内的新血管形成的证据。与rho/VEGE小鼠一样，新生血管起源于视网膜的深毛细血管床，但它更广泛，并引起视网膜外皱褶，随后发生完全视网膜脱离。实时聚合酶链反应和酶联免疫吸附试验表明，与表现出更温和的表型的rho/VEGF小鼠相比，具有可诱导VEGF表达的小鼠发展为视网膜脱离，具有更高的VEGF mRNA和蛋白质的眼部水平。这些数据表明，无论血管床的发育阶段如何，视网膜中VEGF表达的增加足以引起新生血管形成，并且高水平的表达引起严重的新生血管形成和牵引性视网膜脱离。在视网膜中具有可诱导的VEGF表达的小鼠提供了有价值的眼部新生血管形成的新模型。
Transgenic. mice with vascular endothelial growth factor (VEGF) driven by the rhodopsin promoter (rho/VEGF mice) develop neovascularization that originates from the deep capillary bed of the retina and grows into the subretinal space. in rho/VEGF mice, VEGF expression in photoreceptors begins between postnatal days 5 and 7, the period when the deep capillary bed is developing. An important question is whether or not the developmental stage of the deep capillary bed is critical for occurrence of neovascularization. Also, although rho/VEGF mice are extremely useful for the study of ocular neovascularization, there are some applications for which the early onset of VEGF expression is a disadvantage. In this study, we used the reverse tetracycline transactivator (rtTA) inducible promoter system coupled to either the rhodopsin or interphotoreceptor retinoid-binding protein (IRBP) promoter to control the time of onset of VEGF transgene expression in photoreceptors. in the absence of doxycycline, adult double-transgenic rho/rtTA-TRE/VEGF or IRBP/rtTA-TRE/VEGF mice showed little VEGF transgene expression and no phenotype. The addition of doxycycline to the drinking water resulted in prominent transgene expression and evidence of neovascularization within 3 to 4 days. Like rho/VEGE mice, the neovascularization originated from the deep capillary bed of the retina, but it was more extensive and caused outer retinal folds followed by total retinal detachment. Real-time polymerase chain reaction and enzyme-linked immunosorbent assay demonstrated that the mice with inducible expression of VEGF that developed retinal detachment had much higher ocular levels of VEGF mRNA and protein compared to rho/VEGF mice that manifest a much milder phenotype. These data demonstrate that regardless of developmental stage of the vascular bed, increased expression of VEGF in the retina is sufficient to cause neovascularization, and high levels of expression cause severe neovascularization and traction retinal detachment. Mice with inducible expression of VEGF in the retina provide a valuable new model of ocular neovascularization.