Vessel infiltration persists in the adult sema3fa mutant retina. (A–H) Cryosections made through the retinas of Tg(kdrl:mCherry) sema3fa WT (A–C) and mutant (D–F) 5 month old siblings. Nuclei are labeled by Hoechst (white). Blood vessels were detected in the area of the photoreceptor outer segments (os) in the majority of semafaca304 eyes (N = 1, n = 3/4), but not in WT retinas (N = 1, n = 4). C and F are magnified views of the boxed regions in B and E. Yellow chevron points to blood vessel. (G, H) One-year-old adult sema3fa+/+ (G) and sema3fa−/− (H) fish were injected with 2,000,000 MW rhodamine dextran (purple) and fixed 4 hours later. Thick (40 µm) eye cryosections counterstained with nuclear label (Hoechst) reveal leakage of dye (purple, yellow chevron) in the neural retina of semafaca304 fish (H: n = 3/5), but not WT siblings (G: n = 3/3). inl, inner nuclear layer; onl, outer nuclear layer; RGC, retinal ganglion cell layer; RPE/C, retinal pigment epithelium/choroid. Scale bar: 50 µm. Scale bar of inset: 10 µm.
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