Fig. 6-S2

( a) Ablation of Tg(116A:Gal4;UAS:nfsb-mCherry)-labeled neurons. Note that due to sparse expression of the transgenes, ablation of the cH/PVO populations is likely to be partial (<50%). Representative projection images are shown of non-ablated animals (left) and animals following exposure to the chemical MTZ (right, see Materials and methods). Scale bar = 50 μm. Insets (white boxes) show the locations of higher-magnification single-plane images of transgene-labeled cH, aPVO and pPVO areas and neuronal overlap with 5-HT expression (anti-5-HT antibody staining, green color). Scale bar = 20 μm. ( b) Quantification of ablation efficiency. When Tg(116A:Gal4;UAS:nfsb-mCherry) fish were incubated with MTZ, we observed 6.1 ± 0.66 (mean ± SEM) mCherry-positive cells (n = 54 fish). When MTZ was omitted, 31 ± 1.5 cells were mCherry-positive (n = 3 fish). The reduction resulting from ablation was thus ~80% (p = 0.0019, one-tailed Wilcoxon rank-sum test). pPVO (4.3 ± 1.5 control vs 1.4 ± 0.2 ablated, p = 0.0162) and aPVO (8.0 ± 0.6 control vs 1.9 ± 0.3 ablated, p = 0.0015) cells were also affected. Some of the remaining mCherry-positive cells were dimly fluorescent and misshapen/deformed, indicating damage that might impair function. ( c) Similar to Tg(116A:Gal4;UAS:GFP) ( Figure 3—figure supplement 1), there is strong overlap of Tg(116A:Gal4;UAS:nfsb-mCherry) with anti-5-HT immunostaining (green color). Scale bar = 50 μm. Insets (white boxes) show higher-magnification single-plane images of cH, aPVO and pPVO labeling by this transgene and overlap with 5-HT expression. Scale bar = 20 μm. ( d) The Tg(116A:Gal4;UAS:nfsb-mCherry) transgene does not affect feeding in the absence of MTZ, relative to siblings lacking transgene expression. Fed: p = 0.64, n = 11(negative)/10(positive); Dep.: p = 0.91, n = 11(negative)/10(positive), Fed vs Dep.: p = 0.035(negative)/7.7 × 10−4(positive), two-tailed Wilcoxon rank-sum test.