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Vol. 10, Issue 4, 1259-1276, April 1999
and
Departments of *Biomedical Sciences and
Wounding corneal epithelium establishes a laterally oriented, DC
electric field (EF). Corneal epithelial cells (CECs) cultured in
similar physiological EFs migrate cathodally, but this requires serum
growth factors. Migration depends also on the substrate. On fibronectin
(FN) or laminin (LAM) substrates in EF, cells migrated faster and more
directly cathodally. This also was serum dependent. Epidermal growth
factor (EGF) restored cathodal-directed migration in serum-free medium.
Therefore, the hypothesis that EGF is a serum constituent underlying
both field-directed migration and enhanced migration on ECM
molecules was tested. We used immunofluorescence, flow cytometry, and
confocal microscopy and report that 1) EF exposure up-regulated the EGF
receptor (EGFR); so also did growing cells on substrates of FN or LAM;
and 2) EGFRs and actin accumulated in the cathodal-directed half of
CECs, within 10 min in EF. The cathodal asymmetry of EGFR and actin
staining was correlated, being most marked at the cell-substrate
interface and showing similar patterns of asymmetry at various levels
through a cell. At the cell-substrate interface, EGFRs and
actin frequently colocalized as interdigitated, punctate spots
resembling tank tracks. Cathodal accumulation of EGFR and actin did not
occur in the absence of serum but were restored by adding ligand to
serum-free medium. Inhibition of MAPK, one second messenger engaged by
EGF, significantly reduced EF-directed cell migration. Transforming
growth factor Ophthalmology, Institute of Medical Sciences, University
of Aberdeen, Aberdeen AB25 2ZD, Scotland
and fibroblast growth factor also restored
cathodal-directed cell migration in serum-free medium. However, longer
EF exposure was needed to show clear asymmetric distribution of the
receptors for transforming growth factor and fibroblast growth
factor. We propose that up-regulated expression and redistribution of EGFRs underlie cathodal-directed migration of CECs and directed migration induced by EF on FN and LAM.
Corresponding authors. E-mail
addresses: c.mccaig{at}abdn.ac.uk, m.zhao{at}abdn.ac.uk.
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