Purchase this article with an account.
S. He, R. Subramanyan, V. Krasnoperov, D. Zhu, S. Hoffmann, E.A. Barron, A. Rodriguez, S.J. Ryan, P.S. Gill, D.R. Hinton; Soluble Ephb4 Inhibits Platelet Derived Growth Factor Induced Retinal Pigment Epithelial Cell Migration . Invest. Ophthalmol. Vis. Sci. 2006;47(13):1412.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
EphB4 receptor and its ligand (EphrinB2) play an important role in the regulation of cell adhesion, growth and migration. The purpose of this study was to evaluate the effects of soluble EphB4 (sEphB4) receptor on retinal pigment epithelial (RPE) cell migration induced by Platelet derived growth factor–BB (PDGF) in vitro.
Early passage human fetal RPE cells were used in the study. Effect of sEphB4 (0.1–3ug/ml) on PDGF (20ng/ml) induced RPE migration was evaluated using a modified Boyden chamber assay. Attachment was assayed using a modified MTT assay in collagen and fibronectin coated 96–well plates. The pattern of staining for F–actin and focal adhesion kinase (FAK) was analyzed by using phalloidin and anti–FAK specific antibody. The quantitative expression of FAK and activation of MAPK were determined by Western blot after RPE cells were exposed to sEphB4 for 24 hours. The effect of sEphB4 on phosphorylation of EphB4/EphrinB2 was demonstrated by immunoprecipitation.
sEphB4 reduced RPE migration in response to PDGF stimulation. Similarly, sEphB4 inhibited RPE attachment in a dose–dependent manner. F–actin was redistributed to the periphery of the RPE cells after exposure to sEphB4 at a concentration of 3ug/ml. PDGF–induced FAK and MAPK expression was inhibited in the presence of sEphB4 at a dose of 1ug/ml and above.
Decreased RPE cell migration may be related to down regulation of cell attachment, cytoskeletal reorganization and the inhibition of FAK expression and MAPK activation by sEphB4
This PDF is available to Subscribers Only