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Sonic hedgehog induces response of commissural axons to Semaphorin repulsion during midline crossing

Abstract

Pathfinding axons change responses to guidance cues at intermediate targets. During midline crossing, spinal cord commissural axons acquire responsiveness to class 3 Semaphorins and Slits, which regulate their floor plate exit and restrict their post-crossing trajectory into a longitudinal pathway. We found that Sonic Hedgehog (Shh) could activate the repulsive response of pre-crossing axons to Semaphorins. Blocking Shh function with a monoclonal antibody to Shh, 5E1, in 'open-book' explants or by expressing a dominant-negative form of Patched-1, Ptch1Δloop2, or a Smoothened (Smo) shRNA construct in commissural neurons resulted in severe guidance defects, including stalling and knotting inside the floor plate, recrossing, randomized anterior-posterior projection and overshooting after crossing, reminiscent of Neuropilin-2 mutant embryos. Enhancing protein kinase A activity in pre-crossing axons diminished Shh-induced Semaphorin repulsion and caused profound midline stalling and overshooting/wandering of post-crossing axons. Therefore, a morphogen, Shh, can act as a switch of axon guidance responses.

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Figure 1: Sonic Hedgehog induces Semaphorin responsiveness in pre-crossing commissural axons.
Figure 2: Shh function is required for normal midline pathfinding of commissural axons.
Figure 3: Ptch1 is required for normal midline pathfinding of commissural axons.
Figure 4: Smo is required for proper guidance of commissural axons during midline crossing.
Figure 5: Shh may induce Semaphorin responsiveness by reducing the activity of the cAMP/PKA pathway.

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Acknowledgements

We thank V. Castellani for Sema3b cDNA (Universite Claude Bernard), T.M. Jessell for 5E1, S. Brenner-Morton for antibody to HNF-3β (4C7, Developmental Studies Hybridoma Bank), J. Lu for her technical assistance, B. Shafer for encouragement during the course of this study and for critically reading the manuscript, and members of the Zou laboratory for helpful discussions. We also thank the former Neurobiology, Pharmacology and Physiology Department at the University of Chicago, where this study began. This work was supported by a US National Institutes of Health grant to Y.Z. (RO1 NS046357). L.M.P. was supported by the Cellular and Molecular Genetics training grant T32GM7240-31 at the University of California, San Diego.

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L.P. conducted all experiments and wrote the manuscript under the guidance of Y.Z., who supervised the entire project.

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Correspondence to Yimin Zou.

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Parra, L., Zou, Y. Sonic hedgehog induces response of commissural axons to Semaphorin repulsion during midline crossing. Nat Neurosci 13, 29–35 (2010). https://doi.org/10.1038/nn.2457

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