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Knockdown of Fidgetin Improves Regeneration of Injured Axons by a Microtubule-Based Mechanism
Journal article   Open access   Peer reviewed

Knockdown of Fidgetin Improves Regeneration of Injured Axons by a Microtubule-Based Mechanism

Andrew J Matamoros, Veronica J Tom, Di Wu, Yash Rao, David J Sharp and Peter W Baas
The Journal of neuroscience, v 39(11), pp 2011-2024
13 Mar 2019
DOI: https://doi.org/10.1523/JNEUROSCI.1888-18.2018
PMID: 30647150
Featured in Collection :   UN Sustainable Development Goals @ Drexel
url
https://doi.org/10.1523/jneurosci.1888-18.2018View
Published, Version of Record (VoR)Maybe Open Access (Publisher Bronze),  Open
url
https://doi.org/10.1523/JNEUROSCI.1888-18.2018View
Published, Version of Record (VoR) Open

Abstract

Aggrecans - physiology Animals ATPases Associated with Diverse Cellular Activities - genetics ATPases Associated with Diverse Cellular Activities - physiology Axons - physiology Female Ganglia, Spinal - physiology Gene Knockdown Techniques Male Mice, Transgenic Microtubule-Associated Proteins - genetics Microtubule-Associated Proteins - physiology Microtubules - physiology Nerve Regeneration - physiology Neuroglia - physiology Nuclear Proteins - genetics Nuclear Proteins - physiology Rats, Sprague-Dawley
Fidgetin is a microtubule-severing protein that pares back the labile domains of microtubules in the axon. Experimental depletion of fidgetin results in elongation of the labile domains of microtubules and faster axonal growth. To test whether knockdown assists axonal regeneration, we plated dissociated adult rat DRGs transduced using AAV5-shRNA- on a laminin substrate with spots of aggrecan, a growth-inhibitory chondroitin sulfate proteoglycan. This cell culture assay mimics the glial scar formed after CNS injury. Aggrecan is more concentrated at the edge of the spot, such that axons growing from within the spot toward the edge encounter a concentration gradient that causes growth cones to become dystrophic and axons to retract or curve back on themselves. knockdown resulted in faster-growing axons on both laminin and aggrecan and enhanced crossing of axons from laminin onto aggrecan. Strikingly, axons from within the spot grew more avidly against the inhibitory aggrecan concentration gradient to cross onto laminin, without retracting or curving back. We also tested whether depleting fidgetin improves axonal regeneration after a dorsal root crush in adult female rats. Whereas control DRG neurons failed to extend axons across the dorsal root entry zone after injury, DRG neurons in which was knocked down displayed enhanced regeneration of axons across the dorsal root entry zone into the spinal cord. Collectively, these results establish fidgetin as a novel therapeutic target to augment nerve regeneration and provide a workflow template by which microtubule-related targets can be compared in the future. Here we establish a workflow template from cell culture to animals in which microtubule-based treatments can be tested and compared with one another for their effectiveness in augmenting regeneration of injured axons relevant to spinal cord injury. The present work uses a viral transduction approach to knock down from rat neurons, which coaxes nerve regeneration by elevating microtubule mass in their axons. Unlike previous strategies using microtubule-stabilizing drugs, knockdown adds microtubule mass that is labile (rather than stable), thereby better recapitulating the growth status of a developing axon.

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Collaboration types
Domestic collaboration
Web of Science research areas
Neurosciences
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