Hepatocyte growth factor promotes epithelial morphogenesis and occludin linkage to the cytoskeleton in cultured retinal pigment epithelial cells

Citation
N. Yanagihara et al., Hepatocyte growth factor promotes epithelial morphogenesis and occludin linkage to the cytoskeleton in cultured retinal pigment epithelial cells, GR ARCH CL, 239(8), 2001, pp. 619-627
Citations number
42
Categorie Soggetti
Optalmology
Journal title
GRAEFES ARCHIVE FOR CLINICAL AND EXPERIMENTAL OPHTHALMOLOGY
ISSN journal
0721832X → ACNP
Volume
239
Issue
8
Year of publication
2001
Pages
619 - 627
Database
ISI
SICI code
0721-832X(200108)239:8<619:HGFPEM>2.0.ZU;2-9
Abstract
Background: Although hepatocyte growth factor (HGF) is also known as scatte r factor, it induces epithelial morphogenesis in cultured bovine retinal pi gment epithelial (RPE) cells. To elucidate the mechanism of epithelial morp hogenesis, we investigated the influence of HGF on occludin, a major compon ent of tight junctions. Methods: RPE cells were plated on collagen type 1-c oated chamber slides or dishes, 20 ng/ml HGF was added and the cells were i ncubated for 1 week. Cells were harvested at several time-points, and occlu din expression was examined by immunohistochemistry. Detergent extraction p rotocols to identify the intensity of occludin linkage to the cytoskeleton were also used. Occludin expression was determined semiquantitatively by We stern blotting. Results: Fluorescence microscopy revealed that HGF treatmen t increased the levels of insoluble occludin at the cell borders after dete rgent extraction. These level of insoluble occludin and the associated epit helial morphology were maintained for more than 3 weeks after withdrawal of HGF, whereas cells not treated with HGF had a fibroblastic appearance. Wes tern blotting also showed that insoluble occludin was more abundant in HGF- treated cells. Furthermore, immunoreactive bands of insoluble occludin were somewhat larger than those of soluble occludin, suggesting that insoluble occludin may be modified in comparison to soluble occludin. Conclusion: Our results suggest that HGF promotes linkage of occludin to the cytoskeleton. HGF may become a therapeutic candidate in physiological recovery of RPE ce lls and in preparation of RPE monolayers for transplantation.