Studying Axonal Outgrowth and Regeneration of the Corticospinal Tract in Organotypic Slice Cultures
| Autor: | Jürgen Kiwit, Markus Höltje, Friedemann Paul, Anja U. Bräuer, Gudrun Ahnert-Hilger, Martin Pohland, Ulf Strauss, Robert Glumm, Luminita Stoenica, Jana Glumm |
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| Rok vydání: | 2015 |
| Předmět: |
Botulinum Toxins
Green Fluorescent Proteins Pyramidal Tracts Mice Transgenic Biology Green fluorescent protein Mice Organ Culture Techniques Calcium imaging Cell Movement medicine Animals Nerve Growth Factors Spinal cord injury ADP Ribose Transferases Cerebral Cortex Regeneration (biology) Motor Cortex Wild type medicine.disease Spinal cord Actins Axons Nerve Regeneration Mice Inbred C57BL medicine.anatomical_structure Spinal Cord Synapses Corticospinal tract Immunohistochemistry Neurology (clinical) Neuroscience |
| Zdroj: | Journal of Neurotrauma. 32:1465-1477 |
| ISSN: | 1557-9042 0897-7151 |
| DOI: | 10.1089/neu.2014.3467 |
| Popis: | 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 1 d 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 induction. |
| Databáze: | OpenAIRE |
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