Figure 4

(A) Schematic of lineage tracing of neural crest cells in Tg(sox10:CreERT2, myl7:GFP); Tg(ubb:loxP-CFP-loxP-nuc-mCherry); Tg(ins:mCardinal); Et(1121A:GAL4FF); Tg(UAS:GCaMP6s) zebrafish following 5 µM tamoxifen (TM) treatment from 16 to 24 hpf and staining at 108 hpf. (B) Wholemount immunostaining at 108 hpf for mCherry expression (neural crest-derived cells) and counterstaining with DAPI (DNA). Yellow arrowheads point to neural crest-derived cells positive for GCaMP6s expression. (C) Schematic of two-photon ablation experiment. (D) Normalized calcium traces of pancreatic islet cells (including delta, beta, alpha, and unidentified cells). Individual islet cells were assigned to a cell type and given a cell id; LG, basal condition; HG, glucose treated condition; HG a, ablation or mock ablation condition. (E) Correlation matrices of islet cell activity; LG, basal condition; HG, glucose treated condition; HG a, ablation or mock ablation condition. (F) Average beta cell correlation coefficients in individual larvae; n = 11–12 animals, p values from two-way analysis of variance (ANOVA) with Holm–Sidak’s multiple comparisons test. (G) Average homotypic (beta–beta) and heterotypic (alpha–beta) cell correlation coefficients with cell distance distribution from 1 (close) to 10 (far), mean and linear regression (solid lines) with 95% confidence intervals; n = 11–12 animals, p values of slope and intercept from simple linear regression. Fraction time analysis of heterotypic delta–beta, alpha–delta, and alpha–beta cell pairs for times when both are active (H), both are silent (I), and one is active and one is silent (J); mean ± standard error of the mean (SEM), n = 178–215 cell pairs, p values from two-way ANOVA with Holm–Sidak’s multiple comparisons test; magenta circle, active state; cyan circle, silent state.