Eun-Mi Hura,1, In Hong Yangb,1, Deok-Ho Kimb,1, Justin Byuna, Saijilafua, Wen-Lin Xua, Philip R. Nicovicha, Raymond Cheongb, Andre Levchenkob, Nitish Thakorb, and Feng-Quan Zhoua,c,2
aDepartment of Orthopedic Surgery,
bDepartment of Biomedical Engineering, and
cThe Solomon H. Snyder Department of Neuroscience, Johns Hopkins University, Baltimore, MD 21205
Edited by Lynn T. Landmesser, Case Western Reserve University, Cleveland, OH, and approved February 15, 2011 (received for review August 2, 2010)
1E.-M.H., I.H.Y., and D.-H.K. contributed equally to this work.
2To whom correspondence should be addressed.
Neurons in the central nervous system (CNS) fail to regenerate axons after injuries due to the diminished intrinsic axon growth capacity of mature neurons and the hostile extrinsic environment composed of a milieu of inhibitory factors. Recent studies revealed that targeting a particular group of extracellular inhibitory factors is insufficient to trigger long-distance axon regeneration. Instead of antagonizing the growing list of impediments, tackling a common target that mediates axon growth inhibition offers an alternative strategy to promote axon regeneration. Neuronal growth cone, the machinery that derives axon extension, is the final converging target of most, if not all, growth impediments in the CNS. In this study, we aim to promote axon growth by directly targeting the growth cone. Here we report that pharmacological inhibition or genetic silencing of nonmuscle myosin II (NMII) markedly accelerates axon growth over permissive and nonpermissive substrates, including major CNS inhibitors such as chondroitin sulfate proteoglycans and myelin-associated inhibitors. We find that NMII inhibition leads to the reorganization of both actin and microtubules (MTs) in the growth cone, resulting in MT reorganization that allows rapid axon extension over inhibitory substrates. In addition to enhancing axon extension, we show that local blockade of NMII activity in axons is sufficient to trigger axons to grow across the permissive-inhibitory border. Together, our study proposes NMII and growth cone cytoskeletal components as effective targets for promoting axon regeneration.
myelin, glial scar, multi-compartment neuronal culture chamber
Author contributions: E.-M.H. and F.-Q.Z. designed research; E.-M.H. performed the majority of the experiments, I.H.Y. contributed to two-compartment chamber experiments, D.-H.K. performed the time lapse experiment, and S. and W.-L.X. contributed to axon growth experiments; E.-M.H., J.B., W.-L.X., P.R.N. and R.C. analyzed data; E.-M.H. and F.-Q.Z. wrote the paper; and A.L., N.T., and F.-Q.Z. supervised the research.
The authors declare no conflict of interest.
This article is a PNAS Direct Submission.
This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1011258108/-/DCSupplemental.