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Fig. 4
Activation of Neurons with Left dHb Identity Is Correlated with Behavioral Recovery
(A) Confocal images of dHbs in 7- to 8-day-old Tg(gng8:gal4ff); Tg(UAS:GCaMP7a) larvae. Red circles mark neurons that show increased activation post shock, and white circles mark those that are unchanged.
(B) Average change in fluorescence intensity (ΔF/F) for responsive (red) and unresponsive (gray) neurons. Red and black lines represent the mean for all dHb neurons tested (n = 4,195 in 38 situs solitus from eight clutches; n = 1,246 in 18 right isomerized from four clutches; n = 1,068 in 12 left isomerized from four clutches), and the shaded areas represent SEM. Vertical gold lines indicate time of shock.
(C and D) Mean fluorescence intensity (C), calculated as the area under the curve, during 200 s pre- and post-shock intervals for responsive (R) and nonresponsive (NR) neurons. Mean fluorescence was significantly elevated post shock in all configurations (situs solitus, Wilcoxon W = 741, p < 0.0001; right isomerized, Wilcoxon W = 105, p < 0.0001; left isomerized, Wilcoxon W = 105, p < 0.0001). However, far fewer neurons were responsive in right-isomerized dHb (D) (Kruskal-Wallis ANOVA H = 50.4; situs solitus, 20.2% ± 2.0%; right isomerized, 2.4% ± 0.6%, Dunn’s post test, p < 0.0001; left isomerized, 30.0% ± 4.1%, Dunn’s post test, p < 0.0001).
(E) Responsive neurons are more abundant in the left dHb of situs solitus larvae (left; Wilcoxon W = 741, p < 0.0001) but are distributed equally between both dHbs in right- (Wilcoxon W = 1, p > 0.05) and left-isomerized individuals (Wilcoxon W = 81, p > 0.05).
Black circles in (C)–(E) correspond to individual larvae, and red bars represent mean ± SEM superimposed over each scatterplot. Significance below p = 0.05 (∗) is noted. See also Figures S3 and S4 and Movie S2.