FIGURE 3. Decreased PSA-NCAM in dorsal hippocampus of IUGR mice at P18.
Analysis performed in z-stacks captured from double immunolabeling detecting PV IR in green (Alexa 488 goat anti chicken) and PSA-NCAM IR in red (Alexa 568 goat anti mouse), and DAPI nuclear staining in dorsal hippocampus. Box and whiskers plots represent PSA-NCAM total volume (μm3 × 103 μm3; A1, B1, & C1) and total IF (A.U. × 103 μm3; A2, B2, & C2) at P18 in CA1 and CA3. Analysis segmented for Or (A), Rd (B), and Py (C) layers of the dorsal CA1 and CA3. Boxes are limited by the 75th and 25th 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 reconstruction renderings from z-stacks using Zen blue software in CA1 (D1) and CA3 (D2) are shown. The decrease in PSA-NCAM IR (x-axis) inversely correlated with the increase in GAD 65/67 puncta per neuron (E1) and volume per neuron (E2) at P18. Continuous line represents the fitted line derived from a linear regression and the discontinuous line represent the 95% CI. Spearman Rho correlation was applied. A.U., arbitrary units; CA, cornus ammonis; GAD, Glutamic acid decarboxylase; IF, immunofluorescence; Or, Oriens Layer; P, post-natal age; pcta, puncta; PV, parvalbumin; Py, pyramidal cell layer; r, Rho coefficient; Rd, Radiatum layer.