MCT8 regulates axon branching in the Rohon-Beard sensory neurons. A. A representative scheme of the Rohon-Beard (RB) sensory neuron location in zebrafish larvae. B-D. Double fluorescent ISH in 33 hpf embryos revealed co-localization of p2rx3.1 (green) and mct8 (red) in RB cell bodies. E-F. Whole mount ISH showed the spatial expression of p2rx3.1 in the dorsal spinal cord of 2 dpf WT-sibling (E) and mct8-/- larvae (F). G-I. Whole-mount ISH and immunofluorescence revealed co-localization of EGFP (green) and p2rx3.1 (red) in the cell body of an RB neuron in 2 dpf huc:GAL4+uas:memYFP-injected embryos. J. The percentages of embryos that express memYFP in single arborized RB neurons in the tail (black bars), are shown in 2 dpf WT-sibling, mct8-/- and mct8 mRNA-injected mct8-/- embryos. Statistical significance was determined by the Chi square test. Different letters indicate significant difference. K. The percentages of embryos that express memYFP in single arborized RB neurons in the tail (black bars), are shown in 2 dpf WT-sibling, mct8-/-, WT-sibling treated with 0.5 nM TRIAC and mct8-/- treated with 0.5 nM TRIAC. Statistical significance was determined by the Chi square test. Different letters indicate significant difference. L, M. Lateral view of arborized RB-neuron that projects toward the tail in 2 dpf live mct8-/- and WT-sibling embryos, which are transiently expressed huc:GAL4 and uas:memYFP constructs. N. Schematic illustration of arborized RB sensory neuron. Each color represents a single branch that was subjected to ImageJ software analysis. Filopodia are colored in black. The total length (O), average length (P), and number of branches (Q) measured in mct8-/- and WT-sibling embryos. Scale bar = 30 µm. Values represented as means±SEM (standard error of the mean). Statistical significance determined by t-test: Two-sample assuming unequal variances followed by one-sample Kolmogorov-Smirnov test, to assume normal distribution (*p<0.05).
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