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Laminin induced local axonal translation of β-actin mRNA is impaired in SMN-deficient motoneurons

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Abstract

Reduced levels of the SMN (survival of motoneuron) protein cause spinal muscular atrophy, the main form of motoneuron disease in children and young adults. In cultured motoneurons, reduced SMN levels lead to disturbed axon growth that correlates with reduced actin mRNA and protein in growth cones, indicating that anterograde transport and local translation of β-actin mRNA are altered in this disease. However, it is not fully understood how local translation of the β-actin mRNA is regulated in SMN-deficient motoneurons. Here, we established a lentiviral GFP-based reporter construct to monitor local translation of β-actin mRNA. Time-lapse imaging of fluorescence recovery after photobleaching (FRAP) in living motoneurons revealed that β-actin is locally translated in the growth cones of embryonic motoneurons. Interestingly, local translation of the β-actin reporter construct was differentially regulated by various Laminin isoforms, indicating that Laminins provide extracellular cues for the regulation of local translation in growth cones. Notably, local translation of β-actin mRNA was deregulated in motoneurons from a mouse model for the most severe form of SMA (Smn −/− ;SMN2). Taken together our findings suggest that local translation of β-actin in growth cones of motoneurons is regulated by Laminin signalling and that this signalling is disturbed in SMA.

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Acknowledgments

We are grateful to Regine Sendtner and Nicole Elflein for excellent technical assistance and Anna Maria Musti for helpful comments on the manuscript. Author contributions: R.B., M.S. designed research; R.R., R.B. performed research, S.H. supported cloning, N.F. did the statistical analysis for best-fit of FRAP curves, R.R. analyzed data, R.R., R.B., M.S. wrote the paper. This work was supported by grants from the Deutsche Forschungsgemeinschaft SFB581, project B1 and B24 and from the Hermann und Lilly Schilling Stiftung im Stifterverband der Deutschen Wissenschaft.

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  1. Steven Havlicek

    Present address: Interdisciplinary Centre for Clinical Research, Nikolaus-Fiebiger Centre for Molecular Medicine, University Hospital Erlangen, 91054, Erlangen, Germany

Authors and Affiliations

  1. Institute for Clinical Neurobiology, University of Wuerzburg, Building E4, Versbacherstr. 5, 97078, Wuerzburg, Germany

    Reena Rathod, Steven Havlicek, Nicolas Frank, Robert Blum & Michael Sendtner

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  1. Reena Rathod
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Correspondence to Robert Blum.

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Rathod, R., Havlicek, S., Frank, N. et al. Laminin induced local axonal translation of β-actin mRNA is impaired in SMN-deficient motoneurons. Histochem Cell Biol 138, 737–748 (2012). https://doi.org/10.1007/s00418-012-0989-1

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