Figure 2. Neuroepithelial flow drives morphological changes from optic vesicle to optic cup: the role of the optic fissure and the impact on the forming stem cell domain.

(A) Dorsal view on optic cup development over time visualized by mosaic nuclear GFP (H2BGFP) (data are derived from 4D imaging data started at 16.5 hpf, one optical section is provided as Video 4), while the lens-facing neuroepithelium is starting to engulf the developing lens (asterisk), the lens-averted epithelium is largely integrated into the lens-facing epithelium by flowing around the distal nasal and temporal rims (arrows). A white dotted line marks the border between lens-facing and lens-averted epithelium. (B) Scheme showing the key findings of A, the lens (asterisk) facing epithelium is marked with red bars. The lens-averted epithelium, which over time is integrated into the lens-facing epithelium is marked with green dots (except the cells at the edges are additionally marked with a yellow core). In between the last cells, which are integrated into the optic cup, the RPE will form in the lens averted domain. (C) shows the percentage of movements with a considerable share in dorso-ventral direction for the dorsal, central, and ventral area of the developing eye. In the ventral area of the eye, there is significantly more movement in the dorso-ventral axis, than in the central or dorsal area. (D) scheme demonstrating the optic vesicle to optic cup transition (lateral view). Notably, the morphological change from the elongated oval optic vesicle to the hemispheric optic cup is driven mainly by the ventral regions (arrows mark the orientation of epithelial flow) (C and D).

DOI: http://dx.doi.org/10.7554/eLife.05216.007

Figure 2—figure supplement 1. Epithelial flow is independent of cell division.

(A) Retinal cell division was inhibited by application of aphidicolin, a well-established DNA polymerase inhibitor. Aphidicolin efficiently inhibited cell proliferation shown by drastically reduced pHH3 positive nuclei (upper panel, average of 6 pHH3 positive nuclei) compared to the control (lower panel, average of 21 pHH3 positive nuclei) (the optic cup is encircled with a dotted white line, 21.5 hpf). (B) We addressed the epithelial flow of aphidicolin-treated wild-type embryos, injected with H2BGFP RNA at the one cell stage; please see Figure 2A as control. The embryo was preincubated with aphidicolin 5 hr prior to the start of imaging (17 hpf, see also Video 2). The application of aphidicolin did not affect the epithelial flow. As a low level side effect of aphidicolin we observed cell death, in line with previous reports, importantly also not affecting the epithelial flow.

DOI: http://dx.doi.org/10.7554/eLife.05216.008