Knockdown of Zebrafish Blood Vessel Epicardial Substance Results in Incomplete Retinal Lamination

Figure 3

Disruption of zBves affected the dysregulated patterns in the retina. The frozen sections and immunofluorescence staining with different marker antibodies—zn8 for ganglion cells (a), (e), anti-CAII for Müller glia (b), (f), anti-TH for interplexiform neurons (c), (g), and zpr-1 for photoreceptors (d), (h) were performed in eyes of 96 hpf wild-type and zBves MO-injected embryos. Immunofluorescence staining with Crx (i), (j) antibody in the frozen sections was performed in 72 hpf embryos. 37.5% (6/16) of mutant retina showed Crx protein. The ultrastructures of the retina in wild-type (m), (m′) and zBves MO-injected embryos (n), (n′) were analyzed by TEM. Double staining with CtBP and zpr-1 was adopted in frozen crytosections (k), (l). 33.3% (4/12) of mutant retina showed CtBP protein without zpr-1 expression (l). Asterisks indicate the outer segment. Black arrows indicate the inner segment. N, nucleus; S, Scale bar in immunostaining photographs, 50 μm. Scale bar in TEM picture, 1000 nm.