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Fig. 5. Sema3d knockdown increases ipsilateral guidance errors and reduces
midline crossing. (A-I) Confocal projections of DiI-labeled RGC axons in
control (A-C) or Sema3d (D-I) morpholino-injected embryos at 2.5
(A,D,G), 3.5 (B,E,H) and 5 (C,F,I) dpf; injected eyes are on the left. A,D-I
are ventral views; B and C are dorsal views. Sema3d knockdown caused aberrant
projections ipsilaterally (arrows) and anteriorly (arrowheads) from the normal
pathway, and increased the occurrence of axons extending to but not beyond the
midline (asterisks). Inset in G shows an enlargement of a growth cone near
midline. Dotted lines indicate midlines; anterior is up in all panels. Scale
bars: 50 µm in A,D-I; 80 µm in B,C. (J) Summary of guidance and
extension errors following Sema3d morpholino (Sema3d MO) knockdown. At 2.5
dpf: control morpholino, n=30 for all analyses; Sema3d morpholino,
n=38 analyzed for reaching the midline, n=31 analyzed for
presence of growth cones, n=28 analyzed for guidance errors. At 3.5
dpf: control morpholino, n=21; Sema3d morpholino, n=22. At 5
dpf: control morpholino, n=30; Sema3d morpholino, n=36.
*P<0.01; **P<0.001;
***P<0.000001.
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