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Schwann cells are axo-protective after injury irrespective of myelination status in mouse Schwann cell-neuron cocultures.

Published version
Peer-reviewed

Repository DOI


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Authors

Fazal, Shaline V 
Schumacher, Nathalie 
Loreto, Andrea 
Coleman, Michael P 

Abstract

Myelinating Schwann cell (SC)-dorsal root ganglion (DRG) neuron cocultures are an important technique for understanding cell-cell signalling and interactions during peripheral nervous system (PNS) myelination, injury, and regeneration. Although methods using rat SCs and neurons or mouse DRG explants are commonplace, there are no established protocols for compartmentalised myelinating cocultures with dissociated mouse cells. There consequently is a need for a coculture protocol that allows separate genetic manipulation of mouse SCs or neurons, or use of cells from different transgenic animals to complement in vivo mouse experiments. However, inducing myelination of dissociated mouse SCs in culture is challenging. Here, we describe a new method to coculture dissociated mouse SCs and DRG neurons in microfluidic chambers and induce robust myelination. Cocultures can be axotomised to study injury and used for drug treatments, and cells can be lentivirally transduced for live imaging. We used this model to investigate axon degeneration after traumatic axotomy and find that SCs, irrespective of myelination status, are axo-protective. At later timepoints after injury, live imaging of cocultures shows that SCs break up, ingest and clear axonal debris.

Description

Peer reviewed: True


Acknowledgements: We thank Karin Müller, Martin Lenz, and Filomena Gallo at the Cambridge Centre for Advanced Imaging for assistance with microscopy. We thank Aviva Tolkovsky and Alex Clark for technical suggestions. We thank Peter Brophy for the gift of the periaxin antibody. We thank Claire Jacob for helpful suggestions.


Publication status: Published


Funder: Belgian National Fund for Scientific Research F.R.S. - FNRS; doi: http://dx.doi.org/10.13039/501100002661


Funder: University of Cambridge; doi: http://dx.doi.org/10.13039/501100000735

Keywords

Axon degeneration, Coculture, Dorsal root ganglion neuron, Mouse, Myelination, Schwann cell, Wallerian degeneration, Animals, Mice, Rats, Coculture Techniques, Neurons, Schwann Cells, Axons, Animals, Genetically Modified

Journal Title

J Cell Sci

Conference Name

Journal ISSN

0021-9533
1477-9137

Volume Title

136

Publisher

The Company of Biologists
Sponsorship
Medical Research Council (2251399)
Wellcome Trust (UNS38871)
Wellcome Trust (210904/Z/18/Z)
Wellcome Trust (220906/Z/20/Z)