Fig. 3

Endocytic trafficking defects cause rw341 mutant lens phenotypes. (A) 5 dpf lenses of wild type, rw341 mutants and rw341 mutants injected with rabenosyn 5 mRNA. (B) Percentage of transparent lens fiber area in wild type, rw341 mutants and rw341 mutants injected with vps45 or rabenosyn 5 mRNA. rabenosyn 5 mRNA increased transparent area size in rw341 mutants, although the difference was less significant (P=0.058). (C) 5 dpf lenses of wild type, rw341 mutants and rw341 mutants injected with rab5aa, rab5c or rab11a mRNA. (D) Percentage of transparent lens fiber area in wild type, rw341 mutants and rw341 mutants injected with rab5aa, rab5c or rab11a mRNA. All types of rab mRNA increased transparent area size in rw341 mutants, although rab5aa mRNA injection was not significant. (E) Expression of GFP-tagged rab5c and rab7 in wild-type and rw341 mutant lens epithelium at 48 hpf. (F) Expression of GFP-tagged rab11a in wild-type and rw341 mutant lens epithelium at 48 hpf. The left panel provides a schematic drawing of a lens epithelial cell with a nucleus (blue) and recycling endosomes (green). Two different level confocal images indicate GFP-rab11a signals densely located beneath apical surface membranes, which face the lens fiber core (lf) at the posterior-most position along the AP axis. Right panels indicate three neighboring confocal images of wild-type and rw341 mutant lens epithelium. In wild-type lens, strong patchy GFP-rab11a signals are observed in the apical region of lens epithelial cells (arrows) adjacent to lens fiber core. In contrast, there are no GFP-rab11a signals in rw341 mutants. (G-I) Percentage of GFP-tagged rab5c (G), rab7 (H) and rab11a (I)-positive area relative to total apical area. (B,D,G-I) Data are mean±s.d. (B,D) Two-way ANOVA. (G-I) Student's t-test: *P<0.05, **P<0.01, ***P<0.005. Scale bars: 20 µm.