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An essential role for β-actin mRNA localization and translation in Ca2+-dependent growth cone guidance

Nature Neuroscience volume 9, pages 1265–1273 (2006)Cite this article

Abstract

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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Figure 1: Inhibition of protein synthesis blocks BDNF-induced growth cone turning.
Figure 2: Growth cone turning by FLIP of caged Ca2+ depends on protein synthesis.
Figure 3: Spatial distribution of β-actin mRNA and ZBP1 proteins in X. laevis neurons and growth cones.
Figure 4: Asymmetric distribution of β-actin mRNA and ZBP1 in growth cones during response to local BDNF application.
Figure 5: Ca2+-dependent growth cone turning depends on β-actin mRNA–ZBP1 interactions.
Figure 6: Asymmetric increase in β-actin in growth cones by local BDNF application.
Figure 7: Reverse asymmetry of β-actin distribution in growth cones during repulsion induced by a BDNF gradient under PKA inhibition.
Figure 8: Asymmetric activation of the Src family of kinases by BDNF gradients.

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Acknowledgements

We thank C.E. Holt and colleagues (University of Cambridge) for sharing unpublished work and R.H. Singer (Albert Einstein College of Medicine) for the antibody to ZBP1.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.).

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Authors and Affiliations

  1. Department of Neuroscience and Cell Biology, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, 08854, New Jersey, USA

    Jiaqi Yao, Zhexing Wen & James Q Zheng

  2. Department of Cell Biology, Emory University School of Medicine, 615 Michael Street, Atlanta, 30322, Georgia, USA

    Yukio Sasaki & Gary J Bassell

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Contributions

J.Y. performed most of the experiments and analyses in this study. Y.S. designed zipcode antisense and did FISH and immunostaining on β-actin mRNA and ZBP1. Z.W. assisted with the turning assay. G.J.B. contributed to the development of the project and directed the antisense and FISH experiments. J.Q.Z. designed and oversaw the research project, and directed most of the experiments, analyses and writing.

Corresponding author

Correspondence to James Q Zheng.

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Supplementary information

Supplementary Fig. 1

The schematic diagram showing the design of antisense oligonucleotides to the Xenopus β-actin zipcode sequences. (PDF 23 kb)

Supplementary Fig. 2

BDNF induced co-localization of β-actin mRNA and ZBP1 in growth cone filopodia. (PDF 25 kb)

Supplementary Fig. 3

Analysis of β-actin asymmetry in Xenopus growth cones exposed to a BDNF gradient. (PDF 35 kb)

Supplementary Fig. 4

The hypothesized model on spatial regulation of β-actin local synthesis in the growth cone. (PDF 73 kb)

Supplementary Fig. 5

Filopodia elicitation by KCl depolarization depends on protein synthesis and β-actin translocation. (PDF 317 kb)

Supplementary Methods (PDF 99 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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