Figure 3.

Increased stimulation of calcium signaling results in a progression of signaling classes. Wing discs were cultured in chemically defined ZB media with varying concentrations of fly extract (FEX): (A) 0%, (B) 2.5%, (C) 10%, and (D) 40% v/v FEX. These conditions stimulate four classes of Ca²⁺ signaling activity: spikes, ICTs, ICWs, and “fluttering” Ca²⁺ oscillations (Videos S14, S15, S16, S17, S18, and S19). (A–D) Montages from ex vivo time-lapse videos are shown. (A′–D′) Kymographs of the corresponding time-lapse videos along the indicated red lines are shown. Note: the kymograph slice in A was selected in the hinge region to clearly indicate examples of spikes. (E) Schematic of four classes of Ca²⁺ signaling activity is shown. The horizontal line represents the x-y position of the kymograph. (E′) Schematic representation of kymographs taken from (E) is shown. (F and G) Average fraction of time in each class of Ca²⁺ signaling activity for 15% FEX varies by (F) pouch age and (G) pouch size. Wing disks are oriented with anterior to the left and dorsal compartment to the top. AEL indicates days after egg laying. Position and timescale bars, 25 μm and 15 min. For (F and G), p < 0.001 for ordinal logistic regression model versus constant model. n indicates the sample size. To see this figure in color, go online.