Extracellular matrix modulation of endothelial cell shape and motility following injury in vitro.
Extracellular matrix modulation of endothelial cell shape and motility following injury in vitro.
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发表时间：
1985-02
影响因子：
4
通讯作者：
W. Young;I. Herman
中科院分区：
文献类型：
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作者：
W. Young;I. Herman
利用荧光显微镜和亲和纯化抗体，对体外培养的损伤后内皮细胞（EC）细胞质肌动蛋白的形态和功能进行了研究。将牛主动脉EC播种于涂有牛血清白蛋白（BSA）、纤维连接蛋白、I型和III型（间质）胶原、IV型（基底膜）胶原或明胶的玻璃显微镜盖上。在玻璃、玻璃- bsa或细胞外基质覆盖的盖层上生长的EC达到汇流后，机械去除300-400微米的细胞区域，以刺激EC的迁移和增殖。在固定前用荧光显微镜用亲和纯化抗体检测肌动蛋白定位，用延时、相衬视频显微术监测损伤后EC的运动。我们发现EC内应力纤维的数量与运动速率成反比；并且，在玻璃或玻璃- bsa上生长的EC的移动速度大约是在明胶或IV型胶原蛋白上生长的EC的8倍（X速度= 0.5微米/分钟对0.06微米/分钟）。EC在纤维连接蛋白和间质胶原上的运动相似（X速度= 0.2微米/分钟）。这些结果表明，细胞外基质分子调节EC应力纤维的表达，从而在体外损伤后产生细胞骨架和EC运动的改变。此外，在基底膜（IV型）胶原存在的情况下，应力纤维的诱导可能解释了在体内动脉粥样硬化过程中，主动脉EC无法迁移和重新填充受损的内膜区域。
We utilized fluorescence microscopy and affinity-purified antibodies to probe the form and function of cytoplasmic actin in endothelial cells (EC) recovering from injury and grown on extracellular matrices in vitro. Bovine aortic EC were seeded onto glass microscope coverslips that had been coated with either BSA, fibronectin, type I and III (interstitial) collagens, type IV (basement membrane) collagen or gelatin. After EC that had been grown on glass, glass-BSA or extracellular matrix-coated coverslips reached confluence, a 300-400 micron zone of cells was mechanically removed to stimulate EC migration and proliferation. Post-injury EC movements were monitored with time-lapse, phase-contrast videomicrography before fixation for actin localization with fluorescence microscopy using affinity-purified antibodies. We found that the number of stress fibres within EC was inversely proportional to the rate of movement; and, the rates of movement for EC grown on glass or glass-BSA were approximately eight times faster than EC grown on gelatin or type IV collagen (X velocity = 0.5 micron/min versus 0.06 micron/min). EC movements on fibronectin and interstitial collagens were similar (X velocity = 0.2 micron/min). These results suggest that extracellular matrix molecules modulate EC stress fibre expression, thereby producing alterations in the cytoskeleton and the resultant EC movements that follow injury in vitro. Moreover, the induction of stress fibres in the presence of basement membrane (type IV) collagen may explain the failure of aortic EC to migrate and repopulate wounded regions of intima during atherogenesis in vivo.