Fig. 5: Effect of excitotoxicity on TrkB-FL transport to the Golgi complex and regulation by MTFL₄₅₇ action.

Cortical neurons were treated with NMDA for the indicated times and analyzed by immunofluorescence with antibodies specific for the Golgi matrix protein GM130 (green) and TrkB-FL Ct (red); nuclear staining was performed with DAPI (blue). Excitotoxicity was induced in the absence of peptides (A–C) or after preincubation with MTMyc and MTFL₄₅₇ (25 μM, 30 min) (D–F). A, D Representative images obtained by confocal microscopy corresponding to single sections, showing channels fused. Scale bar: 20 μm. B, E Mean PCC values ± SD (0-30 min, n = 4; 60 min, n = 3) for TrkB-FL and GM130 colocalization. Statistical analysis was performed using a generalized linear model followed by a post-hoc Fisher’s LSD test (*P < 0.05, **P < 0.01, compared to basal conditions without peptide or with MTMyc, respectively). C, F Percentage of cells showing, at different treatment times, a PCC ≥ 0.52 (C), the value obtained for TrkB-FL/GM130 colocalization in basal conditions in the absence of peptide, or ≥ 0.50 (F). Statistical analysis was performed by one-way ANOVA followed by a Bonferroni post hoc test (*P < 0.05, compared to basal conditions without peptide or with MTMyc, respectively; 0–30 min, n = 4; 60 min, n = 3). For each independent experiment, a minimum of 80 different neurons were analyzed.