Pard3 expression in the retina and the brain. Immunohistochemistry of wild-type retinal neuroepithelium at 33 hpf (A–C), wild-type brain neuroepithelium at 33 hpf (D–F), retinal neuroepithelium of an anti-pard3 morpholino-treated embryo at 33 hpf (G–I), and a wild-type laminated retina at 5 dpf (J–O) that was stained with either anti-Pard3 antiserum (left column) or Alexa 488-conjugated phalloidin (middle column). The corresponding merged images are shown in the right column. The Pard3 protein localized to the apical region of the retinal and brain neuroepithelia at 33 hpf (A and D, respectively) and partially overlapped the adherens junction-associated actin bundles (B and E, arrows). Some of Pard3 localized slightly more apically to the actin bundles (C, arrowheads and inset). Asterisks indicate the position of the lens (A–C and G–I) or the brain ventricles (D–F). Anti-pard3 morpholinos eliminated the expression of Pard3 in the apical regions (arrowheads) of the retinal epithelium (G and I) and resulted in some mislocalization of the actin bundles (H, arrows). Pard3 localized to three major areas in the retina at 5 dpf (J–O): near or apical to the outer limiting membrane (Panels J and M, arrowhead 1), the outer plexiform layer, which correlates with the synaptic termini of the photoreceptor cells (Panels J and M, arrowhead 2), and two layers in the inner plexiform layer (Panel J, arrowheads 3 and 4). The green–red double cones were localized with the zpr-1 monoclonal antibody (L and O, blue). The section was orientated with the RPE on the top and the lens at the bottom. OPL, outer plexiform layer; IPL, inner plexiform layer. Scale bars indicate 20 μm.
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