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Fig. 5. Tract tracing analysis reveals major pathfinding defects in callosal and
hippocampal axons. (A-O) Dual tract tracing was performed at E17.5
by labelling the callosal projection with DiI in the medial cortex (red label
in the figure) and labelling the hippocampal projection through the fornix and
hippocampal commissure with DiA in the dentate gyrus (green label in the
figure). Coronal sections were counterstained with DAPI (A,F,K). Brains were
analysed from rostral to caudal at the level of the corpus callosum and
hippocampal commissure. (There is no green labelling in C as this section is
rostral to the fornix and HC.) In Robo1 knockout mice, callosal (B,
arrowhead; G and L, arrows) and hippocampal (arrowheads in H and M) axons
formed tight fascicles that did not cross the midline. Furthermore, fascicles
from both the cortex and hippocampal projections overlapped at the midline
(knockout overlay images are in I and N), whereas in wild-type mice these
projections remained completely separate (E,J,O). As observed by
immunohistochemical analysis, tract tracing analysis revealed axons crossing
the midline in the rostral region of this Robo1 knockout brain (arrow
in B; D, overlay). Scale bar: 200 µm.
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