Figure 5. Persistent decrease NPTX2 levels in the CA3 in IUGR mice at P40 with minimal dendritic alteration of PV ⁺ INs.

Analysis performed in z-stacks captured from double immunolabeling detecting PV IR in green (Alexa 488 goat anti chicken) and NPTX2 IR in red (Alexa 568 goat anti rabbit), and DAPI nuclear staining in dorsal hippocampus. Box and whiskers plots represent NPTX2 number of puncta per 10³ μm³ (A₁ & B₁), total volume (μm³ × 10³ μm³; A₂& B₂) and total IF (A.U. × 10³ μm³; A₃ & B₃) at P18 in CA1 and CA3. Analysis segmented for Rd (A), and Py (B) layers of the dorsal CA1 and CA3. Boxes are limited by the 75^th and 25^th percentiles (interquartile range, IQR) and whiskers are limited by the last data point within 1.5 times the IQR from the median (continuous line inside the box), with outliers represented as °. Mann-Whitney U test was applied to compared experimental groups. *, p < 0.05. Unbiased image processing and analysis was performed using Imaris x64 v9.8 software blinded to treatments, sex and time. Distribution of the number of Sholl intersections (mean ± SEM, y-axis) counted away from the soma of PV ⁺ INs (x-axis) suggested modest differences between treatment groups at P40 (C). Representative surface reconstruction renderings from z-stacks using Imaris x64 v9.8 software in CA1 (D₁) and CA3 (D₂), showing reconstructed NPTX2 puncta (red channel) and PV ⁺ INs (green channel). A.U., arbitrary units; CA, cornus ammonis; IF, immunofluorescence; Or, Oriens Layer; P, post-natal age; PV, parvalbumin; Py, pyramidal cell layer; Rd, Radiatum layer.