ZFIN Image: Weinschutz Mendes et al., 2023, Fig. 7

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Figure Caption

Fig. 7 Dopaminergic and microglial phenotypes in scn1lab and dyrk1a mutants

(A) Hypothesis-driven GSEA of DE genes (p < 0.1 and fold-change >1.5) in scn1lab^Δ44/Δ44 and dyrk1aa^Δ77/Δ77dyrk1ab^Δ8/Δ8 mutants using cell-type-specific datasets.⁵⁸^,⁵⁹^,⁶⁰ Cell type markers enriched in upregulated (red) or downregulated (blue) genes are shown. Bubbles are shown only for cell type markers with significant enrichment (p < 0.05). The color intensity and size of each bubble represent the odds ratio. p-values calculated using Fisher’s exact test are shown in each bubble. For the complete GSEA in homozygotes and heterozygotes, see Figure S7A.

(B) Phospho-histone H3 (pH3, green) and acetylated tubulin (AcTub, magenta) immunostained whole brains of scn1lab^Δ44/Δ44 and dyrk1aa^Δ77/Δ77dyrk1ab^Δ8/Δ8 versus wild-type fish at 3 dpf. Dorsal views. FB, forebrain; OT, optic tectum; MHB, midbrain-hindbrain boundary. Scale bar, 50 μm. For quantification by brain region, see Figures S7B–S7E.

(C and D) Total number of pH3+ cells in scn1lab^Δ44/Δ44 (n = 18) versus scn1lab^+/+ (n = 16) (C) and dyrk1aa^Δ77/Δ77dyrk1ab^Δ8/Δ8 (n = 14) versus wild-type fish (n = 19) (D). ^∗∗∗∗p < 0.0001, one-way ANOVA.

(E) Tyrosine hydroxylase (TH, green) and acetylated tubulin (AcTub, magenta) immunostained whole brains of wild-type, scn1lab^Δ44/Δ44, and dyrk1aa^Δ77/Δ77dyrk1ab^Δ8/Δ8 fish at 4 dpf. Scale bar, 50 μm. Ventral views. FB, forebrain; PT, posterior tuberculum.

(F and G) Total number of TH+ cells in scn1lab^Δ44/Δ44 (n = 20) versus scn1lab^+/+ (n = 19) in the forebrain (F) and posterior tuberculum (G) (boxes in [E]). ^∗∗∗∗p < 0.0001, one-way ANOVA. For heterozygous phenotypes, see Figures S7F–S7G.

(H) Neutral red staining of live scn1lab^Δ44/Δ44 and dyrk1aa^Δ77/Δ77dyrk1ab^Δ8/Δ8 versus wild-type fish at 4 dpf. Scale bar, 0.1 mm.

(I and J) Total number of neutral red+ cells in scn1lab^Δ44/Δ44 (n = 17) versus scn1lab^+/+ (n = 20) (I) and dyrk1aa^Δ77/Δ77dyrk1ab^Δ8/Δ8 (n = 16) versus wild-type fish (n = 15) (J). ^∗∗∗∗p < 0.0001, ^∗p < 0.05, one-way ANOVA.

(K) 4C4 (green) and acetylated tubulin (AcTub, magenta) immunostaining of whole brains of dyrk1aa^Δ77/Δ77dyrk1ab^Δ8/Δ8 and wild-type fish at 4 dpf. Scale bar, 50 μm. Dorsal views. FB, forebrain; OT, optic tectum; MHB, midbrain-hindbrain boundary.

(L and M) Total number of 4C4⁺ cells in dyrk1aa^Δ77/Δ77dyrk1ab^Δ8/Δ8 (n = 13) versus wild-type (n = 10) in whole brain (L) and forebrain (M). ^∗∗∗∗p < 0.0001, ^∗∗p < 0.01, one-way ANOVA.

Acknowledgments

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