Figure 4.

Changes in iPUT distribution in hippocampal CA neurons in acute slices exposed to neurotoxic conditions

(A) Changes in iPUT accumulation in pyramidal neurons of CA1 and CA2/3 following exposure to CoCl₂ or NMDA. Left: representative confocal images of acute slices incubated for 30 min with iPUT (yellow) in control conditions or in combination with CoCl₂ or NMDA. Slices are labeled for CA1 pyramidal neurons by WFS1 immunofluorescence (magenta). Nuclei are labeled with DAPI (cyan). StO, stratum oriens; StP, stratum pyramidale; StR, stratum radiatum. Scale bars, 500 μm. Right: quantification of mean iPUT fluorescence intensity in CA1 and CA2/3 pyramidal neurons in control slices and in slices exposed to CoCl₂ or NMDA (n = 3 animals).

(B) Changes in intracellular localization of iPUT in CA1 neurons following exposure to CoCl₂ or NMDA. Left: high-magnification representative confocal images of CA1 neurons in acute slices incubated for 30 min with iPUT (yellow) in control conditions or in combination with CoCl₂ or NMDA. Nuclei are labeled with DAPI (cyan). Scale bars, 10 μm. Right: Pearson’s correlation coefficient quantification of colocalization of iPUT-DAPI (indicating intranuclear localization) in CA1 neurons in slices incubated in control conditions or exposed to CoCl₂ or NMDA (n = 3 animals). Data information: (A and B) data are presented as mean ± SEM. (A) Statistical analysis: two-way ANOVA, no effect of region: F(1,12) = 0.721, p = 0.412; statistically significant effect of treatment F(2,12) = 9.517, p = 0.003; no interaction F(2,12) = 0.807, p = 0.469; ∗p < 0.05, ∗∗∗p < 0.001, post-hoc Bonferroni test. (B) Statistical analysis: one-way ANOVA, statistically significant effect of treatment: F(2,6) = 32.69, p = 0.0006, ∗∗p < 0.01, post-hoc Bonferroni test.