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. Author manuscript; available in PMC: 2023 Jan 1.
Published in final edited form as: Dev Neurosci. 2021 Dec 21;44(4-5):214–232. doi: 10.1159/000521611

FIGURE 2. Increased GAD 65/67 + synaptic boutons at P18 in the mouse model of IUGR.

FIGURE 2.

Double immunolabeling detected PV IR in green (Alexa 488 goat anti chicken) and GAD 65/67 IR in red (Alexa 568 goat anti rabbit), and DAPI staining in dorsal CA1 (A & B) and CA3 (C & D), as shown in panels for P10 sham (A & C) and IUGR (B & D) mice. High magnification insets are shown for GAD 65/67 panels (A2, B2, C2, & D2), with bar representing 10 μm. Box and whiskers plots represent PV total volume (μm3) at P18 (E) and GAD 65/67 at P18 (F) and P40 (H). 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 experiments groups. *, p < 0.05. Unbiased image processing and analysis was performed using Imaris x64 v9.8 software blinded to treatments, sex and time. While no changes were documented in the abundance of PV levels in either CA1 or CA3 at P18, GAD 65/67 total volume (μm3 per 103 μm3, F1), volume per puncta (μm3, F2), and number of puncta per neuron (F3) were increased in IUGR mice vs. sham. Representative renderings for GAD 65/67 reconstruction from z-stacks using Zen blue software in CA1 (G1) and CA3 (G2). Increase in GAD 65/67 total volume (H1) and number of puncta per neuron (H2) resolved by P40. CA, cornus ammonis; GAD, Glutamic acid decarboxylase; Or, Oriens Layer; P, post-natal age; PV, parvalbumin; Py, pyramidal cell layer; Rd, Radiatum layer.