Figure 6. Sexual dysmorphism in disruption of myelination in the CA3 of IUGR mice at P40.

Analysis performed in z-stacks captured from immunolabeling detecting MBP IR in magenta (Alexa 647 goat anti mouse) and DAPI nuclear staining in dorsal hippocampus. Box and whiskers plots represent filament analysis using Imaris software for MBP-stained myelinated axons in the CA1 and CA3, and stratification by sex of CA3 analysis. Analysis included total filament length (μm, A₁), surface area (μm², A₂), and volume (μm³, A₃) as well as filament length per neuron (μm, A₄). 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. Representative rendering of z-stacks produced using ZenBlue software for CA1 (B₁) and CA3 (B₂) of sham and IUGR female mice are shown, with high magnification panels of reconstruction of myelinated MBP ⁺ axons (a). A second set of high magnification panels localized in the Rd are also shown for CA3 (b). MBP ⁺ filament length (μm; y-axis) directly correlated with deficits in NPTX2 volume (μm³; x-axis) at P40 in the CA3. Continuous line represent the fitted line derived from a linear regression and the discontinuous line represent the 95% CI. Spearman Rho correlation was applied. CA, cornus ammonis; Or, Oriens Layer; P, post-natal age; MBP, Myelin basic protein; Py, pyramidal cell layer; r, Rho coefficient; Rd, Radiatum layer.