Fig. 5
Induction of L/M dysfunction, O-GlcNAcylation decrease, and enhanced astrocyte activation and Aβ accumulation by DON treatment. DON (500 ng/g) was administered via single or multiple cycles of intraperitoneal injections: three consecutive DON treatments (DON), 4 days of recovery without DON following the initial treatment (DON (R)), and a total of 5 cycles consisting of 3 days of DON injections with 4 days of recovery in between (DON (5X)). A day after the final cycle of DON injections, the fear context L/M test or brain analysis was conducted. A Schematic experimental design and procedure of DON, DON (R), and DON (5X). B Representative confocal images (× 40) of DAPI (blue), O-GlcNAc (green), and merged are immunofluorescence staining of the Dl region of zebrafish telencephalon. Enlarged images are presented in the white boxes. The graphs represent the quantitative results for each antibody with normalization by the DAPI levels (n = 5 ~ 6/group). Statistical analysis was performed using the Kruskal–Wallis test with the original FDR of Benjamini and Hochberg (*p < 0.05 versus Con, #p < 0.05 versus DON). C Graphs represent the results of fear context L/M test after different schemes of DON injection. They depict the altered crossing times compared to those observed during the first learning session and display the mean ± SEM (n = 8 ~ 17/group). For statistical analysis, a within-group comparison was performed using the Friedman ANOVA test with the original FDR of Benjamini and Hochberg (ap < 0.05, aap < 0.01 versus 1st learning). Between the group comparison, a two-way ANOVA with Tukey’s multiple comparisons test was performed (*p < 0.05, **p < 0.01 versus Con). (D and E) Representative confocal images (40x) of DAPI (blue), GFAP (D, green), Aβ (E, green), and merged immunofluorescence staining in the Dl and Vd regions of zebrafish telencephalon are shown. Enlarged images are presented in the white boxes. The graphs represent the quantitative results for each antibody with normalization by the DAPI levels (n = 5 ~ 6/group). For statistical analysis, the Mann–Whitney test was performed the two-group comparisons, while the Kruskal–Wallis test with the original Benjamini and Hochberg false FDR was used for comparisons involving more than three groups (*p < 0.05, **p < 0.01 versus Con)