Abstract
Semaphorin guidance molecules act through different receptor complexes to activate multiple signaling cascades leading to changes in axonal growth cone behavior and morphology. We describe here approaches for studying the effect of individual Semaphorins on isolated forebrain neurons from mouse embryos and dissecting downstream signaling pathways. These approaches include the production of recombinant Semaphorin ligands, the culture of dissociated primary neurons, the manipulation of gene expression by electroporation in primary neurons, and functional assays to assess axon outgrowth and growth cone collapse.
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Acknowledgments
We thank Axelle Dovonou for providing pictures of collapsed growth cones. The authors’ research is funded by Centre National de la Recherche Scientifique (CNRS), Aix-Marseille Université, grant from Agence Nationale de la Recherche (ANR-12-BSV4-0012-01) to F.M., and Institut Universitaire de France to S.C.
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Chauvet, S., Mire, E., Mann, F. (2017). Characterizing Semaphorin Signaling Using Isolated Neurons in Culture. 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_16
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DOI: https://doi.org/10.1007/978-1-4939-6448-2_16
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