Retinal ganglion cells in cardiovascular disruption model systems. (A,C) Cryosections of doubly-transgenic (cdh5:gal4; UAS:nfsB-mCherry), DMSO-treated (DMSO Control; (A); Met-treated (Endothelial Cell-Depleted; (B); and Met-treated clutchmates (Met Control; (C) at 72 hpf, stained with zn8, which labels DM-Grasp/Neurolin/Alcama, present on neurons growing long axons, and more weakly expressed in neuroepithelial cells. Control retinas show a well-defined layer of retinal ganglion cells within the ganglion cell layer (GCL) (A, arrow), while endothelial cell-depleted retinas display a reduced and disorganized GCL (B). (D,E) Cryosections of normal clutchmates (D) and sih-/- embryos (E) at 72 hpf, stained with zn8. Normal retinas show a clearly stained zn8+ GCL (D) and sih–/– retinas also show clear staining, with the GCL apparently reduced in thickness and slightly disorganized (E). (F,G) Cryosections of normal clutchmates (F) and vlt–/– embryos (G) at 72 hpf, stained with zn8; the GCL of vlt–/– retina appears normal. (H,J) Quantification of fraction of retina occupied by the GCL (GCL/retina ratio), in retinas of endothelial cell-depleted vs. control embryos (H; p < 0.814, Kruskal-Wallis test, post hoc analysis not justified; n = 7 DMSO controls, 8 Met controls, 10 endothelial cell-depleted), sih–/– vs. normal siblings (I; p = 0.147, Mann-Whitney test; n = 12 normal, 11 sih–/–), and vlt–/– vs. normal siblings (J; p = 0.459, Mann-Whitney test; n = 13 normal, 21 vlt–/–). Scale bar (in A, applies to all) = 50 μm.
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