Figure 4  Morphological features of the brains from Zic2kd/+ mice.
(A) Volumetric analysis of the entire brain, lateral ventricle (LV), and hippocampus (HPC). The values for tissue volumes in Zic2kd/+mice are indicated as percentages of the corresponding wild-type values. The values for ratio of volumes in LV/whole brain and HPC/whole brain are also indicated as percentages of the corresponding wild-type values. A total of 15 pairs of Zic2+/+ and Zic2kd/+ mice were subjected to in vivo MRI imaging. *P < 0.05, **P < 0.01, ***P < 0.001 in t-test. Data is presented as means ± SEM. (B) 3D reconstruction of the outer surface of the brains of Zic2+/+ (+/+) and Zic2kd/+ (kd/+) mice (gray) with lateral ventricle (green). Dorsal (left) and anterior-lateral (right) views are indicated. Note the enlarged lateral ventricles and the narrowed interspace between the left and right lateral ventricles containing septum (asterisk). (C) Morphometric analysis. Sections were subjected to acetylcholine esterase staining. (a)–(d) Lines denote the distances measured in each section (a, cerebral cortex thickness; b, corpus callosum thickness; c, medial structure rostral to the hippocampus [fimbria including septofimbrial nucleus or septal triangular nucleus]; d, subfornical organ width). The measurements were done on 15 pairs of the most comparable from the serial sections of adult male Zic2+/+ and Zic2kd/+ mice brains. (D) Morphometric analysis. The lengths are presented as a percentage relative to the corresponding wild-type values. *P < 0.05, **P < 0.01, ***P < 0.001 in t-test. Data is presented as means ± SEM. (E) Morphological abnormalities in the amygdala. The coronal sections from Zic2+/+ (+/+) and Zic2kd/+ (kd/+) mice were subjected to immunostaining with the anti-Zic antibody, toluidine blue (TB), and acetylcholine esterase staining (AchE). The black arrowhead and arrow indicate the Zic-positive cells in the amygdalohippocampal area (AHA) and medial nucleus, respectively. The white arrowheads indicate the enhanced AchE signals in the AHA. AMY, amygdalar complex; BLA, basolateral nucleus of amygdala; COA, cortical nucleus of amygdala; DT, dorsal thalamic nuclei; LA, lateral nucleus of amygdala; MEA, medial nucleus of amygdala; MH, medial habenular nucleus; MN, meningeal membrane. Scale bars, 0.5 mm.