Fig. 8. Gap junctions do not mediate propagation of the injury-induced Ca²⁺ wave. Cells were pretreated and loaded with fluo-3 AM, washed with HEPES-buffered saline and monitored using CLSM. (A) Neither 20 µM -GA nor 2 mM 1-heptanol prevented propagation of the Ca²⁺ wave. (B) Cells incubated with 5-CFDA were photobleached and followed for 30 minutes. Cortical astrocytes displayed refilling, whereas HCE-Ts did not. (C) A linear wound 50 µm wide was made within a confluent region of cells to create an acellular region (region delineated by two horizontal red lines). One hour later, background images were taken (0 s) at 10x magnification, and then a circular wound (depicted by a red oval) was made near the original linear wound (5 s). The injury-induced Ca²⁺ wave propagated in a normal radial pattern (10 s and 20 s), with no apparent difference in intensity or rate. The horizontal white bar represents 100 µm. Results are representative of three independent experiments.