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Fig. 3
Spatial patterning of cones precedes planar polarized Crumbs localization. (A-A?) Retinal margin in a flat-mount preparation, immunolabeled for Crb2b (magenta) and ZO1 (green). (A) Single z-level focal plane of the Crb2b channel at the level of ZO1 staining (OLM) in the pre-column zone. Crb2b ?ladders? (arrow) in the pre-column zone and adjacent columns. (Note that due to curvature of the retinal surface, the equivalent ZO1 level in the central retina at the left is at a deeper focal plane.) Immature UV cone profiles (white dots) in the pre-column zone. (A?) The maximum intensity z-projection of ZO1 labeling. (A?) Merged image. (B-B?) Retinal margin in a flat-mount preparation, immunolabeled for Crb2a (magenta) with the tr?2: tdTomato (green) Red cone marker. (B) Note small rings of strong Crb2a staining between cone columns (inset, arrows). (B?, B?) The intensely-labeled Crb2a+ rings are not co-labeled with tr?2: tdTomato. (C-C?) Immunoreactivity for Crb2a (magenta) with the rh1: EGFP (green) rod marker. (C?, C?) The strongly immunoreactive Crb2a+ rings surround rh1: EGFP+ labeled rod photoreceptors. Scale bars: 10 ?m (A?, B?, C?)