Results of behavioral test and brain gene expression analysis conducted in larvae. (A) Distance moved, (B) time spent moving, and (C) time spent in the edge of the apparatus by larval subjects assayed with the open-field test (measures for each time block are shown). (D) Distance moved in the light startle test (measures for each of the six experimental phases of the assay are shown). (E) Distance moved in the photic entrainment test; data are divided into the light phase (12 h) and the dark phase of the experiment (12 h). Data represent means ± standard errors bars. (F) Expression levels of the selected genes cyp19a1b, pax6a, shank3a, and gad1b in heads of 4 dpf larvae. 60 heads were used for each condition, in three biological replicates. Data in all the panels are presented as means ± SEM * indicates significant differences in transcript levels vs control (DMSO 0.1 %). *p < 0.05; ***p < 0.0005.

Results of behavioral tests conducted in juveniles. (A) Time spent close to the mirror in the sociability test. (B) Lateralization index indicates the preference for observing the social stimulus with the right (positive values) or the left eye (negative values). (C) Distance moved, (D) time spent moving, and (E) time spent on the edge of the apparatus by juvenile subjects assayed with the open-field test. Data represent means ± standard errors bars. * indicates significant differences in the first minute vs control (DMSO 0.1 %). *p < 0.05.

(A) Venn Diagram of the protein differentially expressed in brains of juveniles zebrafish treated with PrP 10 or 1000 μg/L. (B,C) Dotplots of GO terms significantly enriched in the brains of adult zebrafish by PrP 10 μg/L and PrP 1000 μg/L treatments. The size of the points is proportional to the number of differentially expressed proteins annotated in each GO term. The color gradient from red (more significant) to blue (less significant) indicates the level of significance.

Interactomes of the protein differentially expressed in juveniles zebrafish brains treated with PrP 10 μg/L (A) or PrP 1000 μg/L (B). Up-regulated and down-regulated proteins are represented with circles filled, respectively, with shades of red or blue. Magenta borders indicate the most relevant hubs in the networks.

(A) Human phenotypes significantly enriched in juveniles zebrafish brains by PrP 10 μg/L and PrP 1000 μg/L treatments. The size of the points is proportional to the number of featured proteins whereas significance is indicated by the red (more significant) to blue (less significant) gradient. (B, C) Gene-disease association networks in juveniles zebrafish brains induced by PrP 10 μg/L (C) and PrP 1000 μg/L (B) treatments. Disease colors indicate the category: pink for “Mental disorders”, green for “Nervous System Diseases”, and orange for “Behavior and Behavior Mechanisms”. (C) Gene Expression levels of the selected genes shank3a, gad1b, ilvbl, dlg1 lin 60 dpf specimens. Shank3a, gad1b, ilvbl, dlg1l were assessed in 9 brains for each condition of 60 dpf zebrafish. Data are presented as means ± SEM. * indicates significant differences in transcript levels vs control (DMSO 0.1 %). *p < 0.05; **p < 0.005.