Prioritising guidance cues: Directional migration induced by substratum contours and electrical gradients is controlled by a rho/cdc42 switch

Coordinated cell migration is a Fundamental feature of embryogenesis but the intracellular mechanism by which cells integrate co-existing extracellular cues to yield appropriate vectoral migration is unknown. Cells in the cornea are guided by a naturally occurring DC electric field (EF) (electrotaxis) as they navigate non-planar substrata but the relative potencies of electrotaxis and guidance by substratum shape (contact guidance) have never been determined. We tested the hypothesis that vectoral migration was controlled by selective activation of rac, cdc42 or rho in response to a 150 mV/mm EF or to a series of parallel substratum nanogrooves (NGs) 130 nm deep. EFs and NGs were presented singly or in combination. Electrotaxis of dissociated bovine corneal epithelial cells (CECs) on planar quartz required signalling by cdc42 and rho but not rac. Contact guidance by substratum NGs required rho but not cdc42 or rac activities. When an EF and NGs were superimposed in parallel, cathodal electrotaxis along NGs was enhanced compared to that on planar quartz but when they were superimposed orthogonally (vertical NGs with horizontal EF) cells were recruited from contact guidance to electrotaxis, suggesting that the EF was more potent. However, increasing the EF to 250 mV/mm was insufficient to recruit the majority to electrotaxis. Consistent for the cues in isolation, when an EF (150 mV/mm) and NGs were superimposed orthogonally, rac activity was not essential for either contact guidance or electrotaxis. However, attenuation of cdc42 signalling abolished electrotaxis and enhanced contact guidance relative to controls (no drug), whereas inhibiting rho signalling enhanced electrotaxis and rho stimulation enhanced contact guidance. Our data are consistent with the idea that migrating CECs use a cdc42/rho "switch" to sort vectoral cues, with cdc42 controlling electrotaxis and rho controlling contact guidance.