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Methods Mol Biol. 2017;1493:223-235.

Characterizing Semaphorin Signaling Using Isolated Neurons in Culture.

Author information

1
Aix Marseille Univ, CNRS, IBDM, Marseille, France.
2
Aix Marseille Univ, CNRS, IBDM, Marseille, France. fanny.mann@univ-amu.fr.

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.

KEYWORDS:

Axonal growth; Growth cone collapse; Neuronal culture; Semaphorin

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