Fig. 4

Serotonergic and dopaminergic transmission regulates expression of pcp4a (A–E) Graphs showing average pcp4a mRNA fold change in the brains of wild-type fish fed and treated with fluoxetine (A), or food deprived and treated with apomorphine (B), SKF-38393 (C), quinpirole (D), or donepezil (E). mRNA amounts were normalized to control clutchmates. n = 6 in (A) and (D), n = 4 in (B) and (E), n = 3 in (C). n indicates the number of biological replicas (pool of ten brains per replica). (F and G) Scheme (F) and confocal images of a 7 dpf th:gal4-VP16; UAS:EGFP-CAAX larva (G), showing localization of dopaminergic and noradrenergic (locus coeruleus) neuronal clusters. Scale bars in (G), 50 μm. (H) Confocal images of a 7 dpf th:gal4-VP16; UAS:EGFP-CAAX larva immunostained with antibodies against GFP, pERK, and tERK. Scale bars, 50 μm. (I–K) Graphs depicting cumulative percentages of pERK/tERK values in dopaminergic neurons in fed or food-deprived 7 dpf th:gal4-VP16; UAS:EGFP-CAAX larvae. n = 9 larvae (I and J) or n = 10 (K), for each condition. (L) Confocal images of 7 dpf elavl3:gal4; UAS:EGFP larvae, showing localization of pcp4a, drd2a, and drd2b mRNA in GFP-labeled tectal PVNs. Scale bars, 10 μm. (M) Bar graph depicting average percentages of pcp4a+ tectal PVNs expressing either drd2a or drd2b. n = 3 larvae. (N and O) Bar graphs showing average pcp4a mRNA fold change in 5 dpf wild-type fish treated with forskolin (N), or in 7 dpf wild-type larvae treated first with quinpirole for 2 h and then with quinpirole and forskolin for 1 h. mRNA amounts were normalized to control clutchmates. n = 6 (N) or n = 4 (O) biological replicas (pool of ten larvae per replica) per group. ∗ p< 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001; n.s., not significant; one-sample t test (A–E, N, and O [control vs. quinpirole]), two-tailed t test (O, quinpirole vs. quinpirole + forskolin), or two-way ANOVA (I–K). Data in the graphs are shown as mean ± standard error of the mean.