Axon pathfinding requires directional responses of growth cones to extracellular cues, which have been shown to involve local synthesis of protein. The identity and functions of the locally produced proteins remain, however, unclear. Here we report that Ca2+-dependent bidirectional turning of Xenopus laevis growth cones requires localized distribution and translation of β-actin messenger RNA. Both β-actin mRNA and its zipcode-binding protein, ZBP1, are localized at the growth cone and become asymmetrically distributed upon local exposure to brain-derived neurotrophic factor (BDNF). Inhibition of protein synthesis or antisense interference with β-actin mRNA–ZBP1 binding abolishes both Ca2+-mediated attraction and repulsion. In addition, attraction involves a local increase in β-actin, whereas repulsion is accompanied by a local decrease in β-actin; thus, both produce a synthesis- and ZBP1 binding–dependent β-actin asymmetry but with opposite polarities. Together with a similar asymmetry in Src activity during bidirectional responses, our findings indicate that Ca2+-dependent spatial regulation of β-actin synthesis through Src contributes to the directional motility of growth cones during guidance.
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We thank C.E. Holt and colleagues (University of Cambridge) for sharing unpublished work and R.H. Singer (Albert Einstein College of Medicine) for the antibody to ZBP1. This work was supported by grants from the US National Institutes of Health (NS36241 to J.Q.Z. and HD46368 to G.J.B.) and a research grant from New Jersey Commission on Spinal Cord Research (04-3029-SCR to J.Q.Z.).
The authors declare no competing financial interests.
The schematic diagram showing the design of antisense oligonucleotides to the Xenopus β-actin zipcode sequences. (PDF 23 kb)
BDNF induced co-localization of β-actin mRNA and ZBP1 in growth cone filopodia. (PDF 25 kb)
Analysis of β-actin asymmetry in Xenopus growth cones exposed to a BDNF gradient. (PDF 35 kb)
The hypothesized model on spatial regulation of β-actin local synthesis in the growth cone. (PDF 73 kb)
Filopodia elicitation by KCl depolarization depends on protein synthesis and β-actin translocation. (PDF 317 kb)
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Yao, J., Sasaki, Y., Wen, Z. et al. An essential role for β-actin mRNA localization and translation in Ca2+-dependent growth cone guidance. Nat Neurosci 9, 1265–1273 (2006). https://doi.org/10.1038/nn1773
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