Early lens abnormalities in pninavu222 mutants. (A–F) Transverse sections through eyes of normal siblings (A,C,E) and pnina mutants (B,D,F) at different time points, as depicted (n = 10 for each genotype and time point). Asterisks mark the CMZ and arrows point at the ALE. Arrowheads in D point at abnormal regions in the ALE. (G,H) Confocal single-plane images of transverse sections of eyes from normal (G) and mutant (H) 54 hpf embryos, labeled for pHH3 (green) and nuclei (DAPI, blue). Arrowheads and arrows point at pHH3-positive cells in the ALE and CMZ, respectively. (I) Statistical analysis of pHH3 labeling (t-test, one-tailed, error bars show s. e.m). (J,K) Merge of confocal and bright-field images of TUNEL in whole 50 hpf normal (J) and mutant (K) eyes (dorsal view), showing the ALE and lens region. Arrowheads point at TUNEL positive cells and arrows at ALE. (L) Statistical analysis of TUNEL experiment (t-test, one-tailed, error bars show s. e.m). (M,N) Merge of confocal fluorescence and bright-field single-plane images of lenses of Tg(foxe3:nls-EGFP) 3 dpf normal (M) and mutant (N) embryos. Insets show EGFP fluorescence channel alone. EGFP expression (green) is evident in the normal ALE (M, arrows) and in the mutant’s cell mass (N, arrows). EGFP expression is also present in nuclei of surface epithelium (arrowheads). Scale bars in all panels are 20 μm.
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