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. Author manuscript; available in PMC: 2022 Oct 17.
Published in final edited form as: Science. 2021 Apr 9;372(6538):150–156. doi: 10.1126/science.abe7790

Fig. 4. Zic2 is not expressed by ganglion cells in spotted gar and zebrafish.

Fig. 4.

(A to F) Development of the visual system in the spotted gar. All images are 3D light-sheet fluorescence microscopy images of EyeDISCO-cleared spotted gar embryos labeled with Islet1 and acetylated Tubulin. (A and C and E) Top (dorsal) views of spotted gars at 2-3dpf (A), 6-7dpf (C), and 17-18dpf (E). (B and D and F) frontal views of whole spotted gar eyes at 2-3dpf (B), 6-7dpf (D), and 17-18dpf (F). The optic nerve (ON and asterisk) starts to form by 6-7dpf and is well developed by 17-18dpf. The optic chiasm (OC) is formed by 6-7dpf. (G) Coronal cryosection of spotted gar embryos at 17-18dpf labeled for ßIII-tubulin and Islet1. (H to K) cryosection from 17-18dpf spotted gar eyes hybridized with zic2 riboprobe (H and J) and labeled for proliferating cell nuclear antigen (PCNA) and Islet1 (I, K and L). (J to L) higher magnification of the ciliary margin zone (area framed in I). (M to P) 3D rendering of whole-brain viewed from the top of zebrafish injected with Tg(atoh7:Gal4,14UASubc:T2A-eGFP-pA) (M and N) or Tg(atoh7:Gal4,14UASubc:ZIC2-T2A-eGFP-pA (O and P). N and P show segmented ganglion cell projections. (P) A large ipsilateral projection (arrowhead) is seen in the Tg(atoh7:Gal4,14UASubc:ZIC2-T2A-eGFP-pA)-injected fish. Abbreviations: ON, Optic nerve; OC, Optic chiasm; GCL, Ganglion cell layer; INL, Inner nuclear layer; ONL, Outer nuclear layer. Scale bars are 50 μm in (A, D, F, J, K, L, M to P) and 15 μm in (B) and 80 μm in (C, H and I) and 150 μm in (E) and 200μm in (G).