The Journal of Neuroscience, March 28, 2007, ():
Robo1 and Robo2 Cooperate to Control the Guidance of Major Axonal Tracts in the Mammalian Forebrain
J. Neurosci. López-Bendito et al.
27: 3395
Supplemental Data
Files in this Data Supplement:
- supplemental material
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Supplementary Figure 1. Neocortical (NCx) explants from E17.5 GFP transgenic mice showing Slit2 repulsion to cortical axons. A and B, Neocortical explants were co-cultured in collagen for 4 days in vitro either with mock-transfected COS cells (A) or Slit2-transfected COS cells (B). Note the strong repulsion observed as very few neocortical axons grew in the proximal sector in the Slit2 experimental cases. Scale bar equals 200 μm.
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Supplementary Figure 2. Thalamocortical projections are already defective at E14.5 in Robo1;Robo2 double mutant mice. A and B, Coronal sections through the telencephalon of E14.5 wild type (A) and Robo1;Robo2 mutants (B) embryos showing computer-generated overlays of DiI labeled axons and Hoestch counterstain after DiI crystals were implanted in the neocortex (NCx). At this developmental stage the trajectory of corticofugal axons in Robo1;Robo2 double mutant mice was indistinguishable from wild type brains. C and D, Coronal sections through the diencephalon of E14.5 wild type (C) and Robo1;Robo2 mutants (D) embryos showing computer-generated overlays of DiI labeled axons and Hoestch counterstain after DiI crystals were implanted in the dorsal thalamus (dTh). Numerous DiI-labeled thalamocortical axons invaded abnormally the hypothalamus (Hyp) in Robo1;Robo2 double mutant embryos (arrows). LGE, lateral ganglionic eminence. Scale bars: 500 μm (A-B); 1 mm (C-D).
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Supplementary Figure 3. Expression of Robo1 and Robo2 mRNA in Robo2 and Robo1 single mutants. Coronal sections through mid-telencephalic/rostral diencephalic levels of E14.5 embryos showing the expression of Robo1 (A-D) and Robo2 (E-H) in Robo2 +/+ (A and B), Robo2 -/- (C and D), Robo1 +/+ (E and F) and Robo1 -/- (G and H). Dorsal is up, and the midline is to the right in all panels or delineated by a dotted line. A–H, No differences were observed in the level of expression of Robo1 in the Robo2 mutant mice, or vice versa. Hb, habenula; vz, ventricular zone; cp, cortical plate; iz, intermediate zone; NCx, neocortex; LGE, lateral ganglionic eminence; dTh, dorsal thalamus. Scale bars: 100 μm (A,C,E,G); 200 μm (B,D,F,H).
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Supplementary Figure 4. Expression of Robo1 and Robo2 proteins in Robo2 and Robo1 single mutants. Coronal sections through mid-telencephalic levels of E14.5 embryos showing the expression of Robo1 in Robo2 -/- (A and B) and of Robo2 in Robo1 -/- (C and D). A–D, No differences were observed in the level of expression or localization of Robo1 in the Robo2 mutant mice compared to wild type (see Figure 1), or vice versa. ic, internal capsule; MPO, medial preoptic area; vz, ventricular zone; cp, cortical plate; iz, intermediate zone; NCx, neocortex; LGE, lateral ganglionic eminence; dTh, dorsal thalamus. Scale bars: 300 μm (A,C) and 200 μm (B,D).
