Fig. 2

Brain-wide activity maps were generated by using phosphorylated-ERK (pERK) antibody staining as a proxy for neuronal activity. a Individual larval stacks were registered for use with the Z-brain atlas and MAP-mapping MATLAB scripts (Randlett et al. ¹², Engert lab) Individual pERK stacks left were then divided by total-ERK (tERK; middle), providing normalized pERK/tERK signal (right). b Median p/tERK values were then calculated for every voxel within the brain for each genotype and light condition (Exact sample sizes of biologically independent samples for each condition and genotype (n = lights-on/lights-off); wild-type (n = 16/19), shank3abΔN (n = 19/21) and shank3abΔC (n = 16/15) are indicated on group median images in (b) and also apply to (c) and (d). c Mann–Whitney U z-scores were calculated, comparing lights-off and lights-on, with magenta indicating increased activity during the transition to lights-off (e.g. Medulla Oblongata, MO) and green indicating increased activity during the transition to lights-on (e.g. Optic Tectum, TeO). Regions within the brain that are black did not reach the P < 10⁻⁵ cut-off. d In comparison to wild-type, shank3abΔ−/− mutant models respond to the lights-off condition (magenta) with activation of their pineal (P), but fail to show activation in the MO and spinal cord (sc). a–c All images are 20 µm dorsal z-projections. d Whole brain z- and x-projections. Scale bars = 50 µm.