Gordon S R, Wood M
Department of Biological Sciences, Oakland University, Rochester, MI 48309-4401, USA.
Cell Mol Biol (Noisy-le-grand). 1997 May;43(3):329-36.
In the present study we have examined soybean (Glycine max) agglutinin (SBA) binding to cells of the rat corneal endothelium during wound repair. Circular transcorneal freeze injuries were given to the endothelia and the tissues were organ cultured at 37 degrees C in basal media Eagle with 10% serum for up to 72 hrs. SBA failed to bind to the surface of non-injured corneal endothelium, but strongly bound to cells involved in the wound repair process. Punctate surface binding was detected 24 hrs. post-injury, but stronger binding was observed at 48 hrs. after wounding. In this case, binding appeared to be distinctly distributed around the cell periphery. To investigate SBA binding during wound repair, endothelia were cultured in the presence of SBA (100 and 200 micrograms/ml). Cell migration into the wound area, and hence subsequent wound repair, was not affected at these concentrations. However, both concentrations altered cell morphology and microfilament patterns. Phalloidin staining of cells 24 hrs. after injury revealed that microfilaments appeared thinner and less in number. In addition, distinct aggregations of actin-positive material were detected at cell-to-cell contacts. Cells around the tissue periphery do not partake in the repair process but displayed an SBA concentration dependent fragmentation of their circumferential microfilament bundles. At 48 hrs. post-injury, SBA-treated cells within the wound area, unlike their control counterparts, did not exhibit stress fibers. These results suggest that a SBA binding surface component is associated with the reorganization of actin during corneal endothelial wound repair, and that these cells can migrate across their natural basement membrane without the benefit of a highly organized microfilament cytoskeleton.
在本研究中，我们检测了大豆（Glycine max）凝集素（SBA）在大鼠角膜内皮细胞伤口修复过程中与细胞的结合情况。对角膜内皮进行环形经角膜冷冻损伤，然后将组织在含10%血清的伊格尔基础培养基中于37℃进行器官培养长达72小时。SBA未能与未受伤的角膜内皮表面结合，但与参与伤口修复过程的细胞强烈结合。损伤后24小时检测到点状表面结合，但在受伤后48小时观察到更强的结合。在这种情况下，结合似乎明显分布在细胞周边。为了研究伤口修复过程中SBA的结合情况，将内皮细胞在SBA（100和200微克/毫升）存在的情况下进行培养。在这些浓度下，细胞向伤口区域的迁移以及随后的伤口修复均未受到影响。然而，这两种浓度都改变了细胞形态和微丝模式。损伤后24小时对细胞进行鬼笔环肽染色显示，微丝看起来更细且数量更少。此外，在细胞间接触处检测到肌动蛋白阳性物质的明显聚集。组织周边的细胞不参与修复过程，但显示出其周向微丝束的SBA浓度依赖性断裂。在损伤后48小时，伤口区域内经SBA处理的细胞与对照细胞不同，未表现出应力纤维。这些结果表明，一种SBA结合表面成分与角膜内皮伤口修复过程中肌动蛋白的重组有关，并且这些细胞可以在没有高度组织化的微丝细胞骨架的情况下跨越其天然基底膜迁移。
