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Fig. 4. Retroviral expression of Zic3 caused abnormality in retinal axon projection
toward the optic disc. (A) Retroviral constructs. LTR, long terminal repeat;
gag, gene encoding the viral capsid proteins; env, gene encoding viral envelop
protein; pol, gene encoding viral reverse transcriptase. Colored boxes
indicate the inserted GFP (green) and Zic3 (pink) cDNAs. (B-G) Infected at HH
stage 10-11, the retinas were analyzed at E12 by co-staining with an
anti-neurofilament antibody (B-F) and an anti-viral GAG antibody (G). The
axons were viewed from the optic fiber side of the flat-mount retinas. White
arrows indicate the direction towards the optic disc. (B) In control virus
RCAS-GFP-injected retinas, axons appeared normal, projecting straight toward
the center of the retina. In the RCAS-Zic3-injected retinas, however, some
axons showed abnormal trajectories (C-G). Although the infected area
identified by anti-viral GAG antibody (P27) was much larger than the area with
axonal phenotype, the area with phenotype appeared to be more intensely
stained by anti-p27 (* in G). Note the whole area shown in G is infected.
Images in F and G are from the same field. Scale bar: 50 µm.
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