Fig. 5

cdh11 knockdown inhibits retinal differentiation. A–C: Zebrafish embryos were injected with standard control morpholino oligonucleotide (MO, Control columns) or cdh11 MO (Slight, Moderate, and Severe columns). A: In 2 day postfertilization (dpf) embryos, retinal ganglion cells were visualized using whole-mount zn-5 monoclonal antibody staining and confocal imaging. Ventral view projections of sections through the optic nerve show retinal ganglion cells and their axon processes. In cdh11 morphant embryos, the retinal ganglion cell layer was progressively reduced in thickness and numbers of ganglion cells detected. B: In 3 dpf embryos, cryostat sections were immunolabeled using antibodies that detect retinal layers: HuC/D (HU), which detects retinal ganglion cells and inner nuclear layer neurons; zn-5 (Zn5), which detects retinal ganglion cells and their axons; Pax6 (Pax6), which detects cells in the ganglion cell layer and brightly labeled amacrine cells within the inner nuclear layer (arrowhead in Control image); and zpr-1 (Zpr1), which detects double cone photoreceptors. Cdh11 loss-of-function results in a progressive reduction in immunostaining markers for differentiated cell types in the retina. C: In 4 dpf embryos, cryostat sections were immunolabeled using antibodies that detect retinal layers: Pax6 (Pax6), which detects a variety of cells including brightly labeled amacrine cells within the inner nuclear layer (arrowhead in Control image); and zpr-1 (Zpr1), which detects double cone photoreceptors. The older embryos maintain the Cdh11 loss-of-function phenotype, showing reduced labeling of differentiated cell types. Scale bar = 100 μp>