Figure 3

Mechanical injury induces an intracellular Ca ²⁺ wave in corneal epithelial cultures. (A) (i) Baseline image of Fluo-3 AM-loaded cells 10 s prior to injury. Scale bar = 50 μm. (ii) Initiation of a Ca²⁺ wave 2 s after mechanical injury at point marked by *. (iii–vii) Wave propagation extended to neighboring cells outwards from the wound edge and diminished to approximate basal levels by 150 seconds. Red represents the highest and blue the lowest Ca²⁺ concentration. (B) Changes in fluorescence intensity in cells six rows from the wound edge, expressed as ((F - F₀)/F₀) × 100 as described in Methods. The average peak fluorescence intensity was significantly reduced in Pax6 ^+/- cultures compared with Pax6 ^+/+ (*, p = 0.013) and occurred 12 s after injury in wild-type (red arrowhead) compared with 22 seconds in Pax6 ^+/- cultures (black arrowhead). Arrow, wound. (C) No significant difference in the intensity was observed in cell row 2 between Pax6 ^+/+ and Pax6 ^+/-. Intensity in cell row 6 was significantly lower in Pax6 ^+/- cultures compared with Pax6 ^+/+ cultures (*, p < 0.01).