Regeneration capacity is reduced as CNS axons mature. Using laser-mediated axotomy, proteomics and puromycin-based tagging of newly-synthesized proteins in a human embryonic stem cell-derived neuron culture system that allows isolation of axons from cell bodies, we show here that efficient regeneration in younger axons (d45 in culture) is associated with local axonal protein synthesis (local translation). Enhanced regeneration, promoted by co-culture with human glial precursor cells, is associated with increased axonal synthesis of proteins, including those constituting the translation machinery itself. Reduced regeneration, as occurs with the maturation of these axons by d65 in culture, correlates with reduced levels of axonal proteins involved in translation and an inability to respond by increased translation of regeneration promoting axonal mRNAs released from stress granules. Together, our results provide evidence that, as in development and in the PNS, local translation contributes to CNS axon regeneration.

Centre for Reconstructive Neuroscience Institute of Experimental Medicine Czech Academy of Sciences Prague Czech Republic

Department of Biological Sciences University of South Carolina Columbia 29208 SC USA

Department of Functional Genomics Center for Neurogenomics and Cognitive Research VU University Amsterdam De Boelelaan 1085 1081 HV Amsterdam the Netherlands

John van Geest Centre for Brain Repair Department of Clinical Neurosciences University of Cambridge Cambridge UK

MRC Centre for Regenerative Medicine and MS Society Edinburgh Centre Edinburgh bioQuarter University of Edinburgh Edinburgh UK

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