Fig. 8
Dietary zinc supplementation restores synaptic Shank3 density, but not synaptic Shank2 density, in the basal and lateral nuclei of the amygdala (BLA). A Representative images of Shank2 (green) and synapsin1/2 (red) immunolabelling in the LA of WT mice and Tbr1+/− mice fed with normal or high zinc diet. LA: lateral amygdala; BA: basal amygdala. B Synaptic density of GluN1 in Tbr1+/− LA, measured as the density of synapsin1/2-positive GluN1 puncta normalised to the WT30ppm value, is not significantly different, nor was changed by dietary zinc supplementation (sample size, region of interest/animal number: WT30ppm = 31/5, Tbr+/−30 ppm = 20/4, WT150ppm = 19/4, Tbr+/−150 ppm = 23/4). C Representative images of Shank2 (green) and synapsin1/2 (red) immunolabelling in the BA. D No significant change in the synaptic density of Shank2 is observed in BA of Tbr1+/− mice fed with either normal or high zinc diet, when compared to WT30ppm (sample size, region of interest/animal number: WT30ppm = 30/5, Tbr+/−30 ppm = 18/4, WT150ppm = 20/4, Tbr+/−150 ppm = 20/4). E Representative images of Shank3 (cyan) and synapsin1/2 (red) immunolabelling in the LA. F Significantly reduced synaptic density of Shank3 in Tbr1+/− LA is rescued by high dietary zinc (sample size, region of interest/animal number: WT30ppm = 27/5, Tbr+/−30 ppm = 30/5, WT150ppm = 18/4, Tbr+/−150 ppm = 30/5). G Representative images of Shank3 (cyan) and synapsin1/2 (red) immunolabelling in the BA. H Reduced synaptic density of GluN1 in Tbr1+/− BA is restored by high zinc diet (sample size, region of interest/animal number: WT30ppm = 30/5, Tbr+/−30 ppm = 33/5, WT150ppm = 18/4, Tbr+/−150 ppm = 31/5). Each point represents an individual region of interest image in each experimental group. Scale bar = 5 µm. All data represent mean ± standard error of the mean, analysed using nonparametric Kruskal–Wallis test. ns = not significant, *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001