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Studying axonal outgrowth and regeneration of the corticospinal tract in organotypic slice cultures

Item Type:Article
Title:Studying axonal outgrowth and regeneration of the corticospinal tract in organotypic slice cultures
Creators Name:Pohland, M. and Glumm, R. and Stoenica, L. and Höltje, M. and Kiwit, J. and Ahnert-Hilger, G. and Strauss, U. and Bräuer, A.U. and Paul, F. and Glumm, J.
Abstract:Studies of axonal outgrowth and regeneration after spinal cord injury are hampered by the complexity of the events involved. Here we present a simple and improved in vitro approach to investigate outgrowth, regeneration of the corticospinal tract and intrinsic parenchymal responses. We prepared organotypic co-cultures using explants from the motor cortex of postnatal donor mice, ubiquitously expressing green fluorescent protein, and cervical spinal cord from wild type pups of the same age. Our data show that a) motor-cortical outgrowth is already detectable after one day in culture and is source specific; b) treatment with neurotrophin-3 and C3 transferase from Clostridium botulinum significantly enhances axonal outgrowth during the course of cultivation; c) outgrowing axons form synaptic connections, as demonstrated by immunohistochemistry and calcium imaging; and d) migrating cells of motor-cortical origin can be reliably identified without previous tracing and are mostly neural precursors that survive and mature in the spinal cord parenchyma. Thus, our model is suitable for screening for candidate substances that enhance outgrowth and regeneration of the corticospinal tract and for studying the role of endogenous neural precursors after lesion.
Keywords:Axonal Outgrowth, GFP, Motor Cortex, Organotypic Slice Culture, Spinal Cord, Animals, Mice
Source:Journal of Neurotrauma
ISSN:0897-7151
Publisher:Mary Ann Liebert (U.S.A.)
Volume:32
Number:19
Page Range:1465-1477
Date:1 October 2015
Official Publication:https://doi.org/10.1089/neu.2014.3467
PubMed:View item in PubMed

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