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Focal adhesion kinase signaling at sites of integrin-mediated adhesion controls axon pathfinding

Abstract

Extracellular matrix (ECM) components regulate neurite outgrowth in tissue culture and in vivo. Live imaging of phosphotyrosine (PY) signals revealed that Xenopus laevis growth cones extending on permissive ECM substrata assemble adhesive point contacts containing enriched levels of tyrosine-phosphorylated proteins. Whereas focal adhesion kinase (FAK) signaling is dispensable for the assembly of focal adhesions in non-neuronal cells, FAK activity is required for the formation of growth cone point contacts. FAK-dependent point contacts promote rapid neurite outgrowth by stabilizing lamellipodial protrusions on permissive ECM substrata. Moreover, local FAK activity is required for ECM-dependent growth cone turning in vitro, suggesting that FAK may control axon pathfinding in vivo. Consistent with this possibility, proper growth and guidance of Rohon-Beard sensory neurons and spinal commissural interneurons requires FAK activity. These findings identify FAK as a key regulator of axon growth and guidance downstream of growth cone–ECM interactions.

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Figure 1: PY-containing point contacts are regulated by ECM components.
Figure 2: Paxillin is tyrosine-phosphorylated within point contacts.
Figure 3: FAK promotes point contact tyrosine phosphorylation and growth cone motility.
Figure 4: FAK activity is required for point contact formation and stabilization of lamellipodial protrusions.
Figure 5: FAK promotes substratum-dependent growth cone turning.
Figure 6: Targeted labeling of dorsal Xenopus neural tube.
Figure 7: Outgrowth of Rohon-Beard peripheral processes is impaired by loss of FAK signaling.
Figure 8: FAK activity is required for ventral midline crossing of commissural interneurons in the spinal cord.

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Acknowledgements

We thank M. Halloran and members of the Gomez lab for comments on the manuscript. This work was supported by grant NS41564 from the US National Institutes of Health (to T.M.G.) and by a National Science Foundation predoctoral fellowship (to E.R.).

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Correspondence to Timothy M Gomez.

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Supplementary information

Supplementary Fig. 1

FRNK expression and FAKmo loading impair FAK activity and reduce PY levels at the growth cone. (PDF 1504 kb)

Supplementary Video 1

Dual-colored image sequence of a growth cone on LN expressing DsRed (red) and GFP-dSH2 (green). Note the robust GFP-dSH2 enrichment within stable point contacts (indicated by yellow arrowheads) and at the tips of motile filopodia. Time lapse, 30X. (MOV 5671 kb)

Supplementary Video 2

Fluorescence image sequence of a GFP-FRNK expressing Xenopus spinal neuron growth cone on LN. Note long, stable filopodia and dynamic, unstable lamellipodial protrusions. Time lapse, 60X. (MOV 4710 kb)

Supplementary Video 3

Fluorescence image sequence of a GFP-expressing Xenopus spinal neuron growth cone on LN (blue) at an interface with substratum-bound TN. Note that many transient lamellipodial protrusions extend onto the TN region but are rapidly withdrawn, while lamellipodia extending onto the LN region are stabilized and become the new leading edge. At t=610s this growth cone was fixed and subsequently stained for phospho-FAK. Note presence of phospho-FAK puncta only within lamellipodia on LN (arrowheads). Time lapse, 60x. (MOV 7411 kb)

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Robles, E., Gomez, T. Focal adhesion kinase signaling at sites of integrin-mediated adhesion controls axon pathfinding. Nat Neurosci 9, 1274–1283 (2006). https://doi.org/10.1038/nn1762

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