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Fig. 1 The zebrafish ChP show evolutionarily conserved tissue characteristics (A) Diagram of the fChP. Lateral (top) and dorsal (bottom) views. (B) Gt(foxj1b:GFP) labels fChP epithelial cells. Max projection (B1) and single plane (B2) of the fChP showing the folded epithelium (left). Schemes representing the location of (B1)–(E3) (right). (B3) fChP injected with 70 kDa rhodamine B isothiocyanate (RITC)-dextran (magenta) in CSF. (B4) fChP labeled by acetylated-tubulin antibody marking cilia (white arrows) and axons (yellow arrows). (B5 and B6) fChP labeled with 4C4 antibody (arrows: macrophage on basal [yellow] or apical [white] side). (C) Number of 4C4-positive cells in the apical and basal sides of the fChP. (D) Tg(kdrl:GFP) labels endothelial cells in fChP. Max projection (D1) and single plane (D2–D4) of the fChP immunolabeled with 4C4 (arrows: contact with endothelial cells) (D3) and SV2 antibodies (arrows: contact with endothelial [white] or epithelial [yellow] cells) (D4). (E) Hybridization chain reaction (HCR) of fChP for clu (yellow, ChP marker) and plvapa (magenta). Max projection. (E1–E3) Merged and individual channels. (F) Diagram of the hChP. Lateral (top) and dorsal (bottom) views. (G) Gt(foxj1b:GFP) labels hChP. Max projection (G1) and single plane (G2) of the hChP showing the sheet-like epithelium (left). Schemes representing the location of (G1)–(J3) (right). (G3–G5) hChP labeled with 4C4 antibody (arrows: macrophage on basal [yellow] or apical [white] side). (G6 and G7) hChP labeled with acetylated-tubulin antibody (yellow arrows: axons, white arrows: cilia). (H) Number of 4C4-positive cells in the apical and basal sides of the hChP. (I) Tg(kdrl:GFP)-expressing endothelial cells in hChP. Max projection (I1 and I3) and single plane (I2 and I4) of the hChP (white arrow: endothelial cells at the basal side) immunolabeled with acetylated-tubulin (arrows: contact with endothelial [white] or epithelial [yellow] cells) (I3) and SV2 antibodies (arrows: contact with endothelial [white] or epithelial [yellow] cells) (I4). (J) HCR of the hChP with clu (yellow) and plvapa (magenta). (J1–J3) Merged and individual channels. (K) Diagram of the fChP used for transmission electron microscopy. (L) Images of the fChP at low (L1 and L2) and high (L1a and L2a) magnification. (L3–L5) Images of junctions (L3), basal membrane (L4), and motile cilia and microvilli (L5). DAPI labels nuclei. Yellow dotted lines mark either the ChP or ventricles. A, anterior; P, posterior; D, dorsal; V, ventral; R, right; L, left; Hb, habenula; Tel, telencephalon. Scale bars: (A1–J3) 50 μm, (L1–L2a) 1 μm, and (L3–L5) 200 nm. Wilcoxon signed-rank test (C) (n = 7) (H) (n = 5). ∗p < 0.05. See also Figures S1–S4.