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Fig. 5
Perturbed basal lamellipodial activity leads to compromised rim involution and impairs RNE morphogenesis.
(A) Confocal scan of rim cells in mKate2-ras injected ezrin morphant. Arrow indicates basal blebs in rim cells. Scale bar = 5 µm. (B) Brightfield images of side view of 30 ss optic cup. Control (top) and ezrin morphant (bottom). Arrowhead marks the epithelial accumulation outside the RNE. Scale bar = 50 µm. (C) Confocal scan of optic cup at 30 ss stained for phalloidin. Control (top) and ezrin morphants (bottom). Arrows mark the epithelial accumulation outside the RNE. Asterisk marks the lens. Lookup table indicates the minimum and maximum intensity values. Scale bar = 10 µm. (D,E) Normalized average intensity distributions of phalloidin (D) and phosphomyosin (E) in the tissue volume along the apicobasal axis of the RNE at 30 ss. Mean ± SEM. Control (brown) ezrin Mo (magenta). Tissue sections, n = 25 for control and n = 30 for ezrin Mo; N = 5 embryos each. See Figure 5—source data 1, 2. (F) Time-lapse imaging of RNE morphogenesis in ezrin morphant injected mosaically with ras-mKate2 RNA. Arrows mark rim cells that failed to move. Dashed line marks the outline of developing RNE. Frames from Video 18. Time in h:min. Scale bar = 10 µm. Imaging started at 17 ss – 18 ss. (G) Confocal scan of 30 ss optic cup expressing paxillin-mKate2 and GFP-ras. Control (top), ezrin Mo (bottom). Arrow marks paxillin enrichment at the basal side. N = 7. Scale bar = 10 µm. (H) Confocal scan of paxillin-mKate2 and GFP-ras expressing cells. Control RNE cells (top), ectopic RNE cells in ezrin morphant (bottom). Arrow marks basal paxillin enrichment. Scale bar = 5 µm.