CRMP2 deficiency leads to callosal hypoplasia. Shortening of corpus callosum (arrowheads) is apparent in both sagittal (first row) and coronal sections (second row) of adult brains. Scale bars: 1 mm.
Quantification of callosal length in 30‐day‐old mice (P30, n = 3, crmp2
−/− is 80.8 ± 5% of WT, P = 0.003) and adult mice (n = 3, crmp2
−/− is 80.5 ± 3% of WT, P = 0.002), mean ± SD, *P < 0.05, t‐test.
Labeling of adult corpus callosum with anti‐neurofilaments antibody in sagittal sections. Outline depicts missing posterior part of the tract in crmp2
−/− mice. Caudal part of the corpus callosum in WTs is located dorsally above the habenular commissure (hbc), while in crmp2
−/− mice, callosum terminates rostrally before reaching the hbc (arrows). PC indicates posterior commissure. Scale bars: 500 μm.
The growth of GFP‐labeled callosal axons in the contralateral cortex at P6 (embryos were electroporated at E15.5). Note the disorganized paths of crmp2
−/− axons. Scale bars: 200 μm.
Schematic drawing of the callosal axon path and axonal tortuosity calculation. Pyramidal neurons in layer II/III project their axons into the contralateral cortex [rectangle depicts the area displayed in (D)]. Axonal tortuosity quantification as shown in (F), tortuosity = 1 if a = b.
Quantification of tortuosity of axons (WT n = 6 pups, crmp2
−/−
n = 3 pups) upon their exit from the callosal tract (WT 1.016 ± 0.003 vs. crmp2
−/− 1.044 ± 0.012, P < 0.001), mean ± SD, ***P < 0.001, t‐test.
DiI‐labeled callosal axons from P9 oblique brain sections (see the
Appendix Fig S1) and their reconstruction in Neurolucida 360 (WT
n = 6, c
rmp2
−/−
n = 9). Scale bars: 200 μm.
Polar histograms of callosal axons reconstructed in (G). Note the broader range of axon growth angles in crmp2
−/− mice.
Left: schematic representation of polar histogram analysis by clustering the traced axons (WT n = 6 pups, crmp2
−/−
n = 9 pups) into three groups based on the growth angles. Right: proportion of axons growing in selected clusters (WT, 0° to ±20°: 67.3 ± 10.3%; ±20° to ±40°: 21.2 ± 5.3%; ±40° to ±60°: 6.22 ± 3.2%; crmp2
−/−, 58.5 ± 5%, P = 0.047, 27.7 ± 4.3%, P = 0.022, 8.64 ± 2.25%, P = 0.11), mean ± SD, *P < 0.05, t‐test.