Quantitative Analysis of the Effects of Extracellular Matrix Proteins on Membrane Dynamics Associated with Corneal Epithelial Cell Motility
Quantitative Analysis of the Effects of Extracellular Matrix Proteins on Membrane Dynamics Associated with Corneal Epithelial Cell Motility
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DOI：
10.1167/iovs.09-4380
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发表时间：
2010-09-01
影响因子：
4.4
通讯作者：
Nishida, Teruo
中科院分区：
文献类型：
--
作者：
Kimura, Kazuhiro;Kawano, Sumihiro;Nishida, Teruo
目的.角膜上皮细胞与基底膜组分的相互作用调节细胞形态、粘附和迁移。作者研究了基底膜成分对人角膜上皮（HCE）细胞行为的影响，包括与细胞迁移方向相关的细胞膜运动。将HCE细胞铺在纤连蛋白、层粘连蛋白、I型和IV型胶原蛋白以及牛血清白蛋白（BSA）上。通过结晶紫染色评价细胞粘附。通过延时显微镜监测细胞运动，并开发图像处理程序用于定量分析细胞膜运动与细胞迁移方向的关系。肌动蛋白细胞骨架，粘着斑，和K+通道Kv2.1通过荧光显微镜检测。层粘连蛋白，纤连蛋白，胶原蛋白I型和IV型各自促进细胞粘附相比，明显与BSA。纤连蛋白和胶原蛋白I型和IV型，但不是层粘连蛋白，增加细胞的运动。与BSA存在下观察到的情况相比，层粘连蛋白对膜运动没有影响。I型和IV型胶原蛋白各自增加膜运动，但以独立于细胞迁移方向的方式。纤连蛋白诱导膜向细胞迁移方向移动。纤连蛋白诱导形成厚束的F-肌动蛋白和粘着斑在细胞周边和Kv2.1的定位在细胞的前缘，而层粘连蛋白和胶原蛋白I型和IV型引起形成较薄的束的F-肌动蛋白和较小的粘着斑。纤维连接蛋白可调节角膜上皮细胞的定向迁移。（Invest Ophthalmol维斯科学。2010;51：4492- 4499）DOI：10.1167/iovs.09- 4380
PURPOSE. Interaction of corneal epithelial cells with components of the basement membrane regulates cell morphology, adhesion, and migration. The authors examined the effects of basement membrane components on the behavior of human corneal epithelial (HCE) cells, including movement of the cell membrane in relation to the direction of cell migration.METHODS. HCE cells were plated on fibronectin, laminin, collagen types I and IV, and bovine serum albumin (BSA). Cell adhesion was evaluated by staining with crystal violet. Cell movement was monitored by time-lapse microscopy, and an image processing program was developed for quantitative analysis of movement of the cell membrane in relation to the direction of cell migration. Actin cytoskeleton, focal adhesions, and the K+ channel Kv2.1 were detected by fluorescence microscopy.RESULTS. Laminin, fibronectin, and collagen types I and IV each promoted cell adhesion compared with that apparent with BSA. Fibronectin and collagen types I and IV, but not laminin, increased cell motility. Laminin had no effect on membrane movement compared with that observed in the presence of BSA. Collagen types I and IV each increased membrane movement but in a manner independent of the direction of cell migration. Fibronectin induced membrane movement in the direction of cell migration. Fibronectin induced the formation of thick bundles of F-actin and focal adhesions at the cell periphery and the localization of Kv2.1 at the leading edge of the cell, whereas laminin and collagen types I and IV elicited the formation of thinner bundles of F-actin and smaller focal adhesions.CONCLUSIONS. Fibronectin may modulate the directional migration of corneal epithelial cells. (Invest Ophthalmol Vis Sci. 2010;51:4492- 4499) DOI:10.1167/iovs.09- 4380