Fig 7

Fig 7 Th-immunoreactive neurons in the olfactory bulb and preoptic area show increased activity in ap2s1 and pappaa mutants respectively.

(A) Summed Z-projection of Th signal within the olfactory bulb obtained from the Z-brain Atlas [31]. (A’) Representative example of a summed projection from a single experiment showing regions of increased activity within the olfactory bulb in ap2s1 mutants relative to their siblings during habituation learning. (A”) A merge of A and A’ showing co-registration of Th-immunoreactive areas and regions of increased activity in ap2s1 mutants relative to siblings during habituation. Areas of increased activity in ap2s1 mutants during habituation learning show strong overlap with Th-immunoreactive olfactory bulb regions. (B) A single z-plane through the olfactory bulb Th-stained area shown in (A). (B’) The same z-plane as in B taken from the stack in A’ showing regions of increased activity within the olfactory bulb in ap2s1 mutants relative to their siblings during habituation learning. (B”) Merge of (B) and (B’). The x-y overlap indicated in (A-A”) occurs within the same z-plane. (C-C’) same as (A’-A”) but for pappaa mutants. (D-D’) same as (B’-B) but for pappaa mutants. (E) Summed Z-projection of anti-Th signal within a sub-region of the preoptic area obtained from the Z-brain Atlas [31]. (E’) Representative example of a summed projection from a single experiment showing regions of increased activity within the preoptic area in pappaa mutants relative to their siblings during habituation learning. (E”) A merge of E and E’ showing co-registration of Th-stained areas and regions of increased activity in pappaa mutants relative to siblings during habituation. Areas of increased activity in pappaa mutants during habituation learning show strong overlap with Th-positive preoptic regions. (F) A single z-plane through the preoptic area shown in (E). (F’) The same z-plane as in (F) taken from the stack in E’ showing regions of increased activity within the preoptic area in pappaa mutants relative to their siblings during habituation learning. (F”) Merge of (F) and (F’). The x-y overlap indicated in (E’-E”) occurs within the same z-plane. (G-G’) same as (E’-E”) but for ap2s1 mutants. (H-H’) same as (F’-F”) but for ap2s1 mutants. ap2s1 mutants do not show upregulation within neurons of the preoptic area that stain positive for Th. (I) Quantification of the fold-change in pERK/tERK in ap2s1 mutants relative to their siblings within ROIs encompassing either the olfactory bulb dopaminergic area on each side or ROIs encompassing the preoptic dopamine clusters on each side. Each point represents the region-specific fold-change for a single larva. Larvae are pooled from all replicates. Olfactory bulb, n = 61 mutants, Wilcoxon Signed Rank Test comparing to null hypothesis of median 1 for the fold change in pERK/tERK in mutants as compared to siblings, p<0.0001. Preoptic area, n = 56 mutants, Wilcoxon Signed Rank Test comparing to null hypothesis of 1 for the fold change in pERK/tERK in mutants as compared to siblings, p = 0.9436. (J) As in (I) but for pappaa mutants relative to their siblings. Olfactory bulb n = 89 mutants. Wilcoxon Signed Rank Test comparing to null hypothesis of 1 for the fold change in pERK/tERK in mutants as compared to siblings, p<0.0001. Preoptic area n = 88 mutants, Wilcoxon Signed Rank Test comparing to null hypothesis of 1 for the fold change in pERK/tERK in mutants as compared to siblings, p<0.0001. (K) The same data set as in (I) but showing the aggregated fold-change averages within each of the 6 ap2s1 replicates to demonstrate high degree of replicability across experiments. Each point is the pERK/tERK fold-change averaged across all animals for a single replicate. Olfactory Bulb, n = 6 replicates, Bonferroni-adjusted p = 0.0624 trends toward significance. (L) The same data set as in (J) but showing aggregated fold-change averages within each of the 7 pappaa replicates to demonstrate high degree of replicability across experiments. Each point is the pERK//tERK fold-change averaged across all animals for a single replicate. Olfactory bulb and preoptic area, each n = 7 replicates, Bonferroni-adjusted for both p = 0.0468. See S2 Fig for activity decreases within these same regions.