Motor paralyses due to damage to the peripheral and central nervous systems, especially those due to motor nerve damage, cause serious disabilities in ADL (activities of daily living), but no effective treatment has been established for this condition. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Ĭompeting interests: The authors have declared that no competing interests exist. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are creditedĭata Availability: All relevant data are available in the paper and its Supporting Information files.įunding: This study was supported by Grants-in-Aid from the MEXT, MHLW, and AMED of Japan. Received: FebruAccepted: OctoPublished: November 16, 2015Ĭopyright: © 2015 Yagi et al. PLoS ONE 10(11):Įditor: Antal Nógrádi, University of Szeged, HUNGARY (2015) Zonisamide Enhances Neurite Elongation of Primary Motor Neurons and Facilitates Peripheral Nerve Regeneration In Vitro and in a Mouse Model. We propose that zonisamide is a potential therapeutic agent for peripheral nerve injuries as well as for neuropathies due to other etiologies.Ĭitation: Yagi H, Ohkawara B, Nakashima H, Ito K, Tsushima M, Ishii H, et al. At 8 weeks after surgery, zonisamide was protective against denervation-induced muscle degeneration in tibialis anterior, and increased gene expression of Chrne, Colq, and Rapsn, which are specifically expressed at the neuromuscular junction. Zonisamide also improved the sciatic function index, a marker for motor function of hindlimbs after sciatic nerve autograft, from 6 weeks after surgery. In a mouse model of sciatic nerve autograft, intragastric administration of zonisamide for 1 week increased the size of axons distal to the transected site 3.9-fold. Zonisamide induced mRNA expression of nerve growth factors (BDNF, NGF, and neurotrophin-4/5), and their receptors (tropomyosin receptor kinase A and B). Zonisamide was also protective against oxidative stress-induced cell death of primary motor neurons. The neurite-scratch assay revealed that zonisamide enhanced neurite regeneration. We found that zonisamide, an anti-epileptic and anti-Parkinson’s disease drug, promoted neurite elongation in cultured primary motor neurons and NSC34 cells in a concentration-dependent manner. To identify a clinically applicable drug, we screened pre-approved drugs for neurite elongation in the motor neuron-like NSC34 cells. Here we describe assay methods using this fluorescent label, alongside NeuroTrack algorithms, to quantitatively assess changes in neurite morphology over time in either rat primary or human iPSC-derived neuron/astrocyte co-cultures.No clinically applicable drug is currently available to enhance neurite elongation after nerve injury. Utilising a VSV-G pseudotyped lentivirus that encodes a red fluorescent protein (mKate2) driven off a synapsin promoter, neuronal expressionis strengthened and non-neuronal crossover is minimized. As such, we have developed a fluorescent reagent (NeuroLight Red) that specifically labels neurons in co-culture with astrocytes. However, in co-cultures containing neurons and astrocytes, neuronal processes cannot be unambiguously identified and measured using phase images alone. We have previously developed a quantitative mono-culture approach for investigating changes in neurite length and ranching using phase images taken with an IncuCyte ZOOM live-content imaging platform in conjunction with NeuroTrack analysis software. An ideal method to track neurite dynamics would allow continuous automated measurement of structural parameters, including length and number of branch points, in a non-perturbing manner.
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Neurite dynamics can be altered in disease states or following exposure to neurotoxic agents providing a rationale for understanding and quantifying this process (Conforti et al., 2007).