Fig. 9. Conditioned wound media induces a Ca²⁺ response that is inhibited with apyrase. (A) Conditioned media from injured corneal epithelial cells was added to fluo-3-AM-loaded epithelial cells and imaged using confocal microscopy. The response was inhibited when the wound media was treated with 30 U/ml apyrase for two minutes. (B) Cells respond to addition of 2.5 µM ATP or UTP with an immediate increase in [Ca²⁺]_i. Percentage change in average fluorescence was calculated and plotted. (C) No elevation in intracellular Ca²⁺ was detected when ATP was added to non-wounded cells pretreated with thapsigargin. (D) Pretreatment of cells with 50 µM of the phospholipase-C inhibitor, U73122, prevented propagation of a Ca²⁺ wave following injury. Cells immediately adjacent to the injury site (asterisk) display an elevation in intracellular Ca²⁺, and the injury response is similar to that exhibited by cells pretreated with 1 µM thapsigargin (see Fig. 6C). The horizontal white bar represents 50 µm. The intensity scale is shown, with red indicating highest Ca²⁺ levels and blue indicating lowest Ca²⁺ levels. Images are representative of three individual experiments.

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