Neuronal development requires highly coordinated regulation of the cytoskeleton within the developing axon. This dynamic regulation manifests itself in axonal branching, turning and pathfinding, presynaptic differentiation, and growth cone collapse and extension. Semaphorin 3A (Sema3A), a secreted guidance cue that primarily functions to repel axons from inappropriate targets, induces cytoskeletal rearrangements that result in growth cone collapse1. These effects require intra-axonal messenger RNA translation. Here we show that transcripts for RhoA, a small guanosine triphosphatase (GTPase) that regulates the actin cytoskeleton, are localized to developing axons and growth cones, and this localization is mediated by an axonal targeting element located in the RhoA 3′ untranslated region (UTR). Sema3A induces intra-axonal translation of RhoA mRNA, and this local translation of RhoA is necessary and sufficient for Sema3A-mediated growth cone collapse. These studies indicate that local RhoA translation regulates the neuronal cytoskeleton and identify a new mechanism for the regulation of RhoA signalling.
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We thank S. Schlesinger for Sindbis plasmids and N. O'Connor for advice on 3D deconvolution and axonal volume calculation. This work is supported by the National Institute of Mental Health (S.R.J.), the National Alliance for Autism Research (S.R.J.), the Charles A. Dana foundation and the Medical Scientist Training Program (E.Z.M.). A.J. is supported by D. Johnston (Baylor Medical College).
Reprints and permissions information is available at npg.nature.com/reprintsandpermissions. The authors declare no competing financial interests.
This file contains legends for Supplementary Figures S1-S9.
This table contains oligonucleotide sequences used in in situ hybridization and RT-PCR.
Effects of protein synthesis inhibitors on growth cone collapse in severed DRG axons.
In situ hybridization against RhoA and other transcripts in DRG axons.
Isolation of DRG axons using a modified Boyden chamber.
Quantification of in situ hybridization signals in axons and cell bodies.
Effect of increased NGF concentration on growth cone collapse.
Effects of vehicle and anisomycin treatment on the RhoA reporter.
Alignment of the 3′UTRs of RhoA mRNA from different species.
Inhibition of growth cone collapse by C. botulinum C3 exoenzyme.
Infectivity and expression levels of Sindbis-IRES virions.
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