An NGF-responsive element targets myo-inositol monophosphatase-1 mRNA to sympathetic neuron axons

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  • A Corrigendum to this article was published on 01 August 2010

Abstract

mRNA localization is an evolutionary conserved mechanism that underlies the establishment of cellular polarity and specialized cell functions. To identify mRNAs localized in subcellular compartments of developing neurons, we took an original approach that combines compartmentalized cultures of rat sympathetic neurons and sequential analysis of gene expression (SAGE). Unexpectedly, the most abundant transcript in axons was mRNA for myo-inositol monophosphatase-1 (Impa1), a key enzyme that regulates the inositol cycle and the main target of lithium in neurons. A novel localization element within the 3′ untranslated region of Impa1 mRNA specifically targeted Impa1 transcript to sympathetic neuron axons and regulated local IMPA1 translation in response to nerve growth factor (NGF). Selective silencing of IMPA1 synthesis in axons decreased nuclear CREB activation and induced axonal degeneration. These results provide insights into mRNA transport in axons and reveal a new NGF-responsive localization element that directs the targeting and local translation of an axonal transcript.

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Figure 1: SAGE screen of transcripts isolated from cell bodies or axons of sympathetic neurons.
Figure 2: Impa1 mRNA and protein are localized in axons.
Figure 3: Cell bodies and axons of sympathetic neurons express different Impa1 isoforms.
Figure 4: Axonal localization of Impa1-L depends on a previously undescribed NGF-responsive localization element.
Figure 5: NGF-dependent IMPA1-L transport and local translation in axons of SCG explants.
Figure 6: Impa1-L directs localization of reporter vectors containing heterologous 3′ UTRs.
Figure 7: Selective silencing of Impa1-L induces axonal degeneration.
Figure 8: Expression of mouse Impa1-L, but not Impa1-S, rescues the axonal degeneration induced by rat Impa1 silencing.

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GenBank/EMBL/DDBJ

Gene Expression Omnibus

Change history

  • 19 April 2010

    In the version of this article initially published, the last name of the fifth author was misspelled. The correct name is Serena De Vita. The error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We are grateful to R. Kuruvilla, A. Lloyd and M. Raff for insightful comments and to all members of the Riccio laboratory for helpful discussion. CamKII-α-5′myrdGFP3′ expression vector was kindly provided by E. Schuman (California Institute of Technology). We thank A. Garedew for helping with the SAGE assay and statistical analysis. We also thank A. Vaughan for help with microscopy analysis. This work was supported by the UK Medical Research Council (MRC; Research Grant G0500/792) and the European Research Council (Marie Curie International Reintegration Grant MIRG-CT-2005-016501). A.R. is a recipient of a MRC Career Development Award (G117/533) and an MRC Senior Non-Clinical Fellowship (G0802010).

Author information

C.A. helped to design the project, performed most of the experiments, analyzed the data and helped to write the manuscript. C.Z. performed the mRNA targeting assay and contributed to many other experiments throughout the study. R.M. was responsible for the bioinformatics analysis. S.D. and S.F. contributed to the SAGE assay. A.S. analyzed inositol polyphosphate content and helped write the manuscript. A.R., the senior author, designed the project, performed some of the RT-qPCR and axon degeneration experiments, analyzed the data and wrote the manuscript.

Correspondence to Antonella Riccio.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–8, Supplementary Table 1, Supplementary Methods and Supplementary Text (PDF 7371 kb)

Supplementary Movie 1

Time-lapse movie of GFP signal in sympathetic neurons deprived of NGF for 36 hours and exposed to NGF for 12 hours. (MOV 952 kb)

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Andreassi, C., Zimmermann, C., Mitter, R. et al. An NGF-responsive element targets myo-inositol monophosphatase-1 mRNA to sympathetic neuron axons. Nat Neurosci 13, 291–301 (2010) doi:10.1038/nn.2486

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