Electrophoresis of cellular membrane components creates the directional cue guiding keratocyte galvanotaxis

Background: Motile cells exposed to an external direct current electric field will reorient and migrate along the direction of the electric potential in a process known as galvanotaxis. The underlying physical mechanism that allows a cell to sense an electric field is unknown, although several plausible hypotheses have been proposed. In this work we evaluate the validity of each of these mechanisms. Results: We find that the directional motile response of fish epidermal cells to the cathode in an electric field does not require extracellular sodium or potassium, is insensitive to membrane potential, and is also insensitive to perturbation of calcium, sodium, hydrogen, or chloride ion transport across the plasma membrane. Cells migrate in the direction of applied forces from laminar fluid flow, but reversal of electro-osmotic flow did not affect the galvanotactic response. Galvanotaxis fails when extracellular pH is below 6, suggesting that the effective charge of membrane components might be a crucial factor. Slowing the migration of membrane components with an increase in aqueous viscosity slows the kinetics of the galvanotactic response. In addition, inhibition of PI3K reverses the cell's response to the anode, suggesting the existence of multiple signaling pathways downstream of the galvanotactic signal. Conclusions: Our results are most consistent with the hypothesis that electrophoretic redistribution of membrane components of the motile cell is the primary physical mechanism for motile cells to sense an electric field. This chemical polarization of the cellular membrane is then transduced by intracellular signaling pathways canonical to chemotaxis to dictate the cell's direction of travel.