Plexiform layers 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 anti-Synaptic vesicle 2 (SV2), which stains synaptic terminals in inner and outer plexiform layers (IPL, OPL). Control retinas show well-defined plexiform layers (A, arrows), while endothelial cell-depleted retinas display a reduced and disorganized IPL and no clear OPL (B). (D,E) Cryosections of normal clutchmates (D) and sih–/– embryos (E) at 72 hpf, stained with anti-SV2. Normal retinas show well-defined plexiform layers (D), and sih–/– retinas also show well-defined plexiform layers, apparently reduced in thickness, with OPL also limited to the ventral half of the retina. Some of the reduced size appears proportional to the overall reduced eye size (E). (F,G) Cryosections of normal clutchmates (F) and vlt–/– embryos (G) at 72 hpf, stained with anti-SV2; the plexiform layers of vlt–/– retina appear normal. (H,J) Quantification of fraction of retina occupied by the IPL (IPL/retina ratio), in retinas of endothelial cell-depleted vs. control embryos (H; ***p < 0.001, **p < 0.01, Kruskal-Wallis with Conover post hoc; n = 8 DMSO control, 6 Met control, 9 endothelial cell-depleted), sih–/– vs. normal siblings (I; ***p < 0.001, Mann-Whitney test; n = 11 normal, 18 sih–/–), and vlt–/– vs. normal siblings (J; p = 0.0466, Mann-Whitney test; n = 15 for each condition). Scale bar (in A, applies to A-G) = 50 μm.
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