Fig. 5. Ncad regulates outgrowth of amacrine cell dendrites. (A-D) Confocal images of GFP⁺ cell columns in wild-type (A,C) and pac^rw95 (B,D) retinae. White arrows and arrowheads show retinal axons and amacrine cell dendrites, respectively. Irregularly branched neurites extend from the INL in pac^rw95 (B,D, red arrowheads). (E,F) GFP (green) and anti-Pax6 antibody (red) labeling of wild-type (E) and pac^rw95 (F) retinae. In the wild-type retinae, amacrine cell dendrites spread horizontally at the interface between RGCs and amacrine cells (white arrowheads). By contrast, Pax6-positive amacrine cells extend abnormally arborized neurites in the amacrine and RGC layers in pac^rw95 (white arrowheads). There is no equivalent arborization in the bipolar cell layer. (G,H) Confocal images showing morphology of bipolar cells (G) and a RGC (H) in 72 hpf pac^rw95 eyes. White arrows indicate axons of RGCs. (I,J) Anti-PKC labeling (red) of 4 dpf wild-type (I) and pac^rw95 retinae (J). (K) Section of a 3 dpf pac^rw95 eye in which wild-type cells have incorporated into the retina. The wild-type column of cells (brown) forms a normal IPL. The red arrow indicates a wild-type cell positioned within the mixed RGC/amacrine cell layer. (L) Section of a 3 dpf wild-type eye incorporating pac^rw95 cells. The red arrow shows disruption of IPL formation by mutant cells. The red arrowheads indicate isolated mutant amacrine cells showing normal neurite morphology. a, amacrine cells; b, bipolar cells/cell layer; inl, inner nuclear layer; ipl, inner plexiform layer; le, lens; pe, pigmented epthelium; pl, photoreceptor layer; rgc, retinal ganglion cells; rgl, retinal ganglion cell layer.

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