Fig. 4

Network-engineered polytherapy restores resting state brain connectivity in mutants. a Diagram depicting brain areas with statistically significant absolute correlation values (unpaired two-tailed Student's t-test; p < 0.05 considered significant; normality of datasets was tested using the Jarque–Bera normality test) in wild-type larvae at 5 dpf (n = 10). Only these connections are shown in b–e. b–e Diagrams illustrating alterations in functional brain connectivity in scn1lab mutant larvae with respect to wild-type sibling controls under the following conditions: b untreated mutants (DMSO), c mutants treated with fluoxetine alone at the 100% dose, d mutants treated with mifepristone alone at the 100% dose, and e mutants treated simultaneously with fluoxetine and mifepristone using one of the optimal polytherapy dose regimens (both drugs at 30%). For all diagrams, brains are shown from the left side (left), from the dorsal side (center), and from the right side (right). Connections which increase or decrease with respect to wild-type larvae are indicated with blue or red lines, respectively. The thickness of the lines in b–e indicates the magnitude of deviation from normal wild-type connectivity. Functional connectivity for each pair of brain areas is determined at 4+ h post exposure by calculating the average absolute correlation between active supervoxels within each area during the pre-stimulus states. For each compound, the highest dose (100%) corresponds to the concentration used in the initial light-sheet-based screen (see Supplementary Table 1). Ten larvae were analyzed per condition. Ce, cerebellum; HBl, hindbrain left; HBr, hindbrain right; OBl, olfactory bulb left; OBr, olfactory bulb right; OTl, optical tectum, left; OTr, optical tectum, right; Pa, pallium; SPa, sub-pallium; Ha, Ha, habenula; Th, thalamus (For ease of illustration, the areal labels/borders are not always shown on anatomically correct positions/scales)