Abstract
The guidance of axons within the developing nervous system is orchestrated by a variety of cues that successively and complementary attract or repel axons to achieve a stereotyped wiring of neural circuits. Here we present a version of a method that has been widely used to identify and characterize the effect of guidance cues on the orientation of axons. We describe the coculture, within a three-dimensional environment, of dorsal spinal cord explants together with a cell aggregate secreting a candidate cue and the method to quantify the effect of this cue on axon orientation.
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References
Raper J, Mason C (2010) Cellular strategies of axonal pathfinding. Cold Spring Harb Perspect Biol 2:a001933
Bashaw GJ, Klein R (2010) Signaling from axon guidance receptors. Cold Spring Harb Perspect Biol 2:a001941
Vitriol EA, Zheng JQ (2012) Growth cone travel in space and time: the cellular ensemble of cytoskeleton, adhesion, and membrane. Neuron 73:1068–1081
Kolodkin AL, Tessier-Lavigne M (2011) Mechanisms and molecules of neuronal wiring: a primer. Cold Spring Harb Perspect Biol 3:a001727. doi:10.1101/cshperspect.a001727
Nawabi H, Castellani V (2011) Axonal commissures in the central nervous system: how to cross the midline? Cell Mol Life Sci 68:2539–2553
Charron F, Stein E, Jeong J et al (2003) The morphogen sonic hedgehog is an axonal chemoattractant that collaborates with netrin-1 in midline axon guidance. Cell 113:11–23
Kennedy TE, Serafini T, de la Torre JR et al (1994) Netrins are diffusible chemotropic factors for commissural axons in the embryonic spinal cord. Cell 78:425–435
Brose K et al (1999) Slit proteins bind Robo receptors and have an evolutionarily conserved role in repulsive axon guidance. Cell 96:795–806
Butler SJ, Dodd J (2003) A role for BMP heterodimers in roof plate-mediated repulsion of commissural axons. Neuron 38:389–401
Dickson BJ, Gilestro GF (2006) Regulation of commissural axon pathfinding by slit and its Robo receptors. Annu Rev Cell Dev Biol 22:651–675
Evans TA, Bashaw GJ (2010) Axon guidance at the midline: of mice and flies. Curr Opin Neurobiol 20:79–85
Knöll B, Weinl C, Nordheim A et al (2007) Stripe assay to examine axonal guidance and cell migration. Nat Protoc 2:1216–1224
von Philipsborn AC et al (2006) Growth cone navigation in substrate-bound ephrin gradients. Development 133:2487–2495
Gil V, del Río JA (2012) Analysis of axonal growth and cell migration in 3D hydrogel cultures of embryonic mouse CNS tissue. Nat Protoc 7:268–280
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Delloye-Bourgeois, C., Moret, F., Castellani, V. (2017). Performing Axon Orientation Assays with Secreted Semaphorins and Other Guidance Cues. In: Terman, J. (eds) Semaphorin Signaling. Methods in Molecular Biology, vol 1493. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6448-2_17
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DOI: https://doi.org/10.1007/978-1-4939-6448-2_17
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6446-8
Online ISBN: 978-1-4939-6448-2
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