Fig. 3: Anemonefish gja5b expression and transjunctional current assays.
From: Cell-cell communication as underlying principle governing color pattern formation in teleost fishes
A RNA-sequencing of scale samples enriched for orange xanthophores (marker genes: plin 6 and tt39b), black melanophores (marker genes: dct and pmela) and white iridophores (marker genes: apoDa and fhl2a) revealed highest gja5b expression in association with iridophores. Scales were collected from wild-type A. ocellaris, Black A. ocellaris mutant and wild-type A. percula (the latter two fish were added to obtain samples of pure black color). The color scale corresponds to z-scored expression values (z-score = value in each sample – “mean across samples”, all divided by the gene expression standard deviation across samples). B The expression of gja5b was monitored by GFP. The GFP gene driven by gja5b promoter was introduced into 19 wild-type fish (for construct see Fig. S5B). Predominant GFP expression was found in white bar areas (18/19 individuals). C–E Transjunctional current assays in Xenopus oocytes were performed to test the formation and functionality of gap junctions. C In comparison to transmitted current at −100mV for wild-type (WT) Gja5b, little to no current was transmitted by E42K-mutated gja5b, singly or in combination with wild-type. NC, no-construct control. Data are represented as box and whisker plots with boxes containing medians and extending from 25th percentile to 75th percentile (interquartile range) and whiskers extending to the last point within 1.5x interquartile range. Source data are provided as a Source Data file. (Two-sided Overall Kruskal-Wallis, χ2 approximation=24.6, p < 0.0001; shared letters over groups indicate medians not significantly different from one another by post hoc Steel-Dwass comparisons.) D Homotypic gap junction of Gja5b, Gja2a, Gja4a, and Gjc2 are functionally active. E Heterotypic gap junctions between Gja5b and Gja2a and between Gja5b and Gja4a are functionally active.
