FIGURES AND TABLES
FROM:
Expression of disrupted in schizophrenia 1 (DISC1) protein in the adult and developing mouse brain indicates its role in neurodevelopment
I L Schurov, E J Handford, N J Brandon and P J Whiting
BACK TO ARTICLE
Figure 1.
Digital photomicrographs of adult mouse brain showing specificity of polyclonal D27 antibody. (a, c) DISC1-ir (brown) in the cortex and hippocampus, respectively, on the paraffin-embedded sections of mouse brain. (b, d) Preabsorption of D27-ir with the immunizing peptide. Sections were counterstained with Mayers haematoxylin (blue). The immunoreactivity seen in the cortex and hippocampus (a, c) was clearly preabsorbed by the immunizing peptide (b, d).
Full figure and legend (127K)
Figure 2.
Digital photomicrographs of adult mouse brain showing the distribution of DISC1-ir (brown) in different brain regions: (a, b) in cortex, (c, d) in hippocampus, (e, f) in hypothalamus, (g, h) in cerebellum and (i, j) in brain stem. Photomicrographs in the right-hand panel (b, d, f, h, j) show enlarged view of boxes on the left-hand panel photomicrographs (a, c, e, g, i). Sections were counterstained with Mayers haematoxylin (blue). DISC1-ir of different density was observed in all studied areas of the brain. In the cortex (a, b) DISC1-ir was present in all layers with higher intensity in layers II–III, V–VI. In hippocampus (c, d), DISC1-ir was monitored in all regions with a higher intensity in CA3 regions. In hypothalamus (e, f), DISC1-ir was high through whole region. 3V—third ventricle, ARC—arcuate hypothalamic nucleus. In cerebellum, Purkinje cells (PCL) and also some other cells in the granule cell layer (GL) and molecular layer (ML) were DISC1-ir (arrows). In brain stem area (i, j), isolated neurons showed strong DISC1-ir. Sp5—spinal trigeminal tract; Sp5N—spinal trigeminal nucleus. Scale bars, 100 
m (a, c, e, g, i), and 30 m (b, d, f, h, j).
Figure 3.
Confocal images of adult mouse brain demonstrating DISC1-ir in neurons in different brain regions. (a) Colocalization of DISC1-ir (red) with MAP 2 (green) in the cortex, (b) hippocampus, (c) anterior hypothalamus. (d) Colocalization of DISC1-ir (red) with GFAP (green) in hippocampus. DISC1 expression colocalized with MAP2 (notice yellow colour in high power images in a–c). At the same time, DISC1 and GFAP did not demonstrate any coexpression (d). Scale bars, 50 m (a), 80 m (b–d), and 100 m (single labelling images in a–d).
Figure 4.
Confocal images of adult mouse cortical brain sections demonstrating colocalization of DISC1-ir with various neurochemical markers. (a) DISC1-ir (green) and EAACA-ir (red); (b) DISC1-ir (green) and calbindin-ir (red); (c) DISC1-ir (green) and GAD-ir (red); (d) DISC1-ir (green) and gephyrin-ir (red). DISC1-ir was present in all tested type of neurons, in neurons expressing EAAC1 (a), calbindin (b), GAD (c) and gephyrin (d) (notice yellow colour). Scale bars, 20 m (a–d).
Figure 5.
Distribution of DISC1 expression in the developing embryonic mouse brain. (a) Lysates from the pool of whole embryonic mouse brains at different time points (E10.5–E20.5) were blotted for D27 antibody to reveal DISC1-specific bands. Lysate from adult cortex was used as a positive control. (b) The same samples were blotted for 
-tubulin. (c) Quantification of immunoblots from three independent experiments are shown as four lines representing four different DISC1 bands, (d) or were plotted as mean
SEM of band intensity (*
P<0.0175) measured using the Li-Cor Odyssey™. Two doublet bands were identified (71 and 75 kDa; 100 and 105 kDa) in all analysed samples (a). Quantitations of immunoblots demonstrated significant changes in the expression of the band 2 (100 kDa) during development with a peak at E13.5 (c, d).
Figure 6.
Distribution of DISC1 expression in the postnatal mouse brain. (a) Lysates from the pool of a whole mouse brains at different time points (P1 to 6 months old) were blotted for D27 antibody to reveal DISC1-specific bands. (b) Same samples were blotted for -tubulin. (c) Quantification of immunoblots from three independent experiments are shown as four lines representing four different DISC1 bands, (d) or were plotted as meanSEM of band intensity (**
P<0.005) measured using the Li-Cor Odyssey™. Two doublet bands were again identified (71 and 75 kDa; 100 and 105 kDa) in all analysed postnatal samples (a). Quantitations of immunoblots demonstrated changes in the expression of the band 2 (100 kDa) during postnatal period with a significant increase at PD35 that remained high through to 6 months of age (c, d).
Figure 7.
Digital photomicrographs of embryonic mouse brain showing the distribution of DISC1-ir (brown) in the whole embryo at E10.5 (a) and in the embryo head at E14.5 (b) and E17.5 (c). Sections were counterstained with Mayers haematoxylin (blue). DISC1-ir can be seen in different areas, especially in developing cortex (neopallial cortex) and in developing hypothalamus (see high power images). DISC1 expression profile changed through early developmental stages from very low expression level at E10.5 (a) to high expression level in very discreet areas at E14.5 (b) and to adult like pattern of expression at E17.5 (c). Insets in (b) demonstrate a higher power illustration of developing cortex (top) and the olfactory area (bottom) at E14.5. Inset in (c) demonstrates a higher power illustration of developing cortex at E17.5. nc, neopallial cortex; h, hypothalamus; p, pons; oa, olfactory area; iz, intermidiate zone; mz, marginal zone; cp, cortical plate; sp, subplate. Scale bars, 20 m (a), 1 mm (b), 50 m (b, top inset), 30 m (b, bottom inset), 1 mm (c) and 70 m (c, inset).
