ZFIN Image: Coolen et al., 2020, Figure 4

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Figure 4

NGPs Behave Like Early Embryonic Progenitors

(A) Experimental design.

(B) Immunostaining for GS (green) and Sox2 (blue) combined with EdU detection (magenta) on a DMSO control 16 dph killifish pallium. Shown in (B’) is a high magnification of the dorsal view of the 3D reconstruction shown in (B). In (B’’) is 5 μm transverse section through the 3D reconstruction shown in (B). White arrows in (B’’) point to EdU⁺ cells in the parenchyme that we identified as neurons. Scale bars, 50 μm in (B) and (B’’) and 20 μm in (B’).

(C) Proportion of EdU-labeled cells with NGP (blue, Sox2⁺GS⁻), neuronal (yellow, Sox2⁻GS⁻), or RG (green, Sox2⁺GS⁺) identity. ^∗Corrected p value < 0.05. Data were analyzed for each cell type with a two-way ANOVA, followed by pairwise comparisons using Holm’s procedure. Proportions were arcsine transformed prior to analysis. Data are represented as mean ± SEM; n = 2 (t1) and n = 3 (t2) for each treatment condition for each treatment condition at t1 and t2, respectively.

(D) Number of EdU-labeled cells per counted area with NGP (blue, Sox2⁺GS⁻), neuronal (yellow, Sox2⁻ and GS⁻) or RG (green, Sox2⁺GS⁺) identity. ^∗Corrected p value < 0.05. Data were analyzed for each cell type with a two-way ANOVA, followed by pairwise comparisons using Holm’s procedure. Data was rank transformed prior to analysis. Data are represented as mean ± SEM; n = 2 (t1) and n = 3(t2) for each treatment condition.

(E) ISH for Notch1 orthologs (notch1a + notch1b), Dll1 orthologs (dlA + dlD), and neurog1 (magenta) combined with GS immunostaining (green) at 8 dph. Images are high magnifications of the pallial surface on a single optical z-plane. Scale bar, 20 μm.