FIGURE 1

FIGURE 1

Simultaneous imaging of spontaneous activities in hair cells and supporting cells. (A) Cartoon of a neuromast organ illustrating sensory hair cells (magenta) and the surrounding supporting cells (green). (B1–B1′) High-resolution images taken using two arms of our diSPIM system show a representative, day 3, double transgenic neuromast expressing cytoRGECO1 (hair cells, numbered in yellow) and cytoGCaMP6s (supporting cells, a subset is numbered in white, underscored). A top-down view (B1) and the corresponding side view of its orthogonal projection (B1′) clearly show the hair cells and the surrounding supporting cells at day 3. (B2) Temporal curves of spontaneous calcium activity in hair cell 5 (labeled in B1–B1′) and its three surrounding supporting cells (labeled in B1′) within the 15 min recording window show distinct time courses. (B3) Temporal curves of spontaneous calcium activity in the cluster of six hair cells (labeled in B1) reveals distinct response profiles. (B4) Temporal curves of spontaneous calcium activity in the seven supporting cells (labeled in B1) reveals some correlation and synchronization among neighboring supporting cells. (C) Comparisons of Pearson’s R of spontaneous calcium activities at day 3 in neighboring supporting cells, neighboring hair cells and hair cells with its surrounding hair cells, n = 6 neuromasts for HC-HC or SC-SC R values and n = 4 neuromasts for HC-SC R values. (D–D′) Box plots showing quantification of the average magnitude (D) and frequency (D′) of spontaneous calcium activity in immature (day 3) and mature (day 6) hair cells. (E–E′) Box plots showing quantification of the average magnitude (E) and frequency (E′) of spontaneous calcium activity in supporting cells in mature (day 6) and immature (day 3) neuromasts. Each dot in (D–E′) represents one neuromast. An unpaired t-test was used in (D–E′). ****p < .0001. Scale bar = 5 μm.