Fig. 4
From: Olfactory bulb anomalies in KBG syndrome mouse model and patients
Loss of Ankrd11 in E14.5 NPCs leads to decreased proliferation and aberrant neurogenesis in P15 RMS. Also see Supplemental Fig. 3. A Ankrd11fl/fl;RosaYFPSTOP/STOP dams mated with Ankrd11fl/fl;RosaYFPSTOP/STOP;NestinCreERT2 males were injected with TMX at E14.5 and resulting pups were injected with BrdU at P14. Resulting mice (Ankrd11control and Ankrd11nscKO) were collected at P15. Images were collected from anatomically matched medial (M) and lateral (L) sagittal sections encompassing OB. RMS indicated in green. B Schematic of areas measured in P15 RMS analysis. Region 1 encompassed the caudodorsal (CD) zone in the RMS, region 2 the ventral intermediate zone in the RMS, and region 3 where the RMS meets the OB. Black box shows analysis area of region 1 in C. C Representative images of Ankrd11control and Ankrd11nscKO P15 RMS region 1 immunostained for BrdU (pink) and SOX2 (green). Open white arrow indicates BrdU + SOX2- cells, closed white arrow indicates BrdU-SOX2 + cells, and yellow open arrow indicates BrdU + SOX2 + cells. Please see Fig. S3A for approximate image locations. D–J Quantification of C for BrdU + cell number (D) and density (E), SOX2 + cell number (F) and density (G), BrdU + SOX2 + cell number (H) and density (I) and proliferative index of Sox2 + cells (% BrdU + SOX2 + /SOX2 + cells) (J). K Representative images of Ankrd11control and Ankrd11nscKO P15 RMS region 1 immunostained for PSA-NCAM (light blue) and CC3 (green). RMS outline in yellow dashed line is defined by the PSA-NCAM boundaries. White arrows indicate CC3 + cells. Counterstained with Hoechst in white. Please see Fig. S3C for approximate image locations. L–M Quantification of (K) for CC3 + cell number (L) and density (M). N Representative images of Ankrd11control and Ankrd11nscKO P15 RMS region 1 immunostained for PSA-NCAM (light blue) and NEUN (red). RMS outline in yellow dashed line is defined by the PSA-NCAM boundaries. Counterstained with Hoechst in white. PSA-NCAM image is also presented in Fig. S3C inset, along with a precise image location in the OB sections. O–P Quantification of (N) for NEUN + cell number (O) and density (P). Error bars represent SEM. Data was analyzed using unpaired t-test. ns not significant, *p < 0.05 **p < 0.01, ***p < 0.001. n = 4–5 mice per genotype from at least two independent litters. Scale bars represent 100 μm (C, K), 200 μm (N). E embryonic day, L lateral, LV lateral ventricle, M medial, OB olfactory bulb, P postnatal day, RMS rostral migratory stream