Figure 1.

Density of perineuronal nets (PNNs) is decreased in the medial prefrontal cortex (mPFC) of glutamate decarboxylase (GAD)67^+/GFP offspring with prenatal stress (PS). (A) Coronal slices were collected in the prefrontal area (Bregma +1.5 mm to +2.0 mm) and images were obtained from the medial prefrontal area indicated by a red rectangle (left). Representative photomicrographs show the distribution of Wisteria Floribunda Agglutinin (WFA)-labeled PNNs in the mPFC. Fluorescence images of a coronal section in control GAD67^+/GFP offspring and those with PS at P21 immunostained with WFA (red) and (parvalbumin PV, blue). (B) Quantitative analysis of cell density in the mPFC.WFA+ and WFA+/PV+ cells were significantly decreased in mPFC in subjects with PS compared with controls (HT-CTRL: 22 slices; HT-PS: 17 slices from six mice, t-test, *P < 0.05; **P < 0.01). The number of WFA−/PV+ cells was also significantly decreased in mPFC in subjects with PS (t-test, ***P < 0.001). No significant differences were detected in the number of WFA+/PV− in the mPFC between the two groups (t-test, P = 0.344). (C) The reduction rate of PV+ and WFA+/PV+ cells in the mPFC of HT-PS were normalized to the average value obtained from HT-CTRL. Regression analyses revealed a significant linear relationship between reduction rate of PV+ cells and reduction rate of WFA+/PV+ cells in the mPFC of HT-PS (R² = 0.683, P < 0.01). (D) Quantitative analysis of fluorescence intensity of WFA in the mPFC of control and stressed mice. There are no significant differences in WFA intensity between subjects with PS (102 PV+ cells and 78 PV− cells from six mice) and normal control subjects (93 PV+ cells and 82 PV− cells from six mice, 2-tailed t-test). Scale bars: 100 μm. Error bars represent the SEM.