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The Journal of Neuroscience, February 2, 2005, 25(5):1113-1121; doi:10.1523/JNEUROSCI.3931-04.2005

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Development/Plasticity/Repair
Application of Rho Antagonist to Neuronal Cell Bodies Promotes Neurite Growth in Compartmented Cultures and Regeneration of Retinal Ganglion Cell Axons in the Optic Nerve of Adult Rats

Johanne Bertrand,¹ Matthew J. Winton,¹ Nieves Rodriguez-Hernandez,³ Robert B. Campenot,² and Lisa McKerracher^1,3

¹Département de Pathologie et biologie cellulaire, Université de Montréal, Montréal, Québec, Canada, H3T 1J4, ²Department of Cell Biology, University of Alberta, Edmonton, Alberta, Canada, T6G 2H7, and ³BioAxone Thérapeutique, Montréal, Québec, Canada, H2X 3P9

Inactivation of Rho promotes neurite growth on inhibitory substrates and axon regeneration in vivo. Here, we compared axon growth when neuronal cell bodies or injured axons were treated with a cell-permeable Rho antagonist (C3-07) in vitro and in vivo. In neurons plated in compartmented cultures, application of C3-07 to either cell bodies or distal axons promoted axonal growth on myelin-associated glycoprotein substrates. In vivo, an injection of C3-07 into the eye promoted regeneration of retinal ganglion cell (RGC) axons in the optic nerve after microcrush lesion. Delayed application of C3-07 promoted RGC growth across the lesion scar. Application of C3-07 completely prevented RGC cell death for 1 week after axotomy. To investigate the mechanism by which Rho inactivation promotes RGC growth, we studied slow axonal transport. Reduction in slow transport of cytoskeletal proteins was observed after axotomy, but inactivation of Rho did not increase slow axonal transport rates. Together, our results indicate that application of a Rho antagonist at the cell body is neuroprotective and overcomes growth inhibition but does not fully prime RGCs for active growth.

Key words: axon regeneration; optic nerve injury; sympathetic neuron; compartmented culture; RGC survival; slow axonal transport

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Received Sep 22, 2004; revised December 13, 2004; accepted December 14, 2004.

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