Targeting chondroitinase ABC to axons enhances the ability of chondroitinase to promote neurite outgrowth and sprouting


Autoři: Priscilla Day aff001;  Nuno Alves aff002;  Esther Daniell aff001;  Debayan Dasgupta aff001;  Rosalie Ogborne aff001;  Ashley Steeper aff001;  Mansoor Raza aff001;  Clare Ellis aff001;  James Fawcett aff002;  Roger Keynes aff001;  Elizabeth Muir aff001
Působiště autorů: Department of Physiology Development and Neuroscience, University of Cambridge, Cambridge, United Kingdom aff001;  John Van Geest Centre for Brain Repair, University of Cambridge, Cambridge, United Kingdom aff002
Vyšlo v časopise: PLoS ONE 15(1)
Kategorie: Research Article
doi: 10.1371/journal.pone.0221851

Souhrn

Background

There is currently no effective treatment for promoting regeneration of injured nerves in patients who have sustained injury to the central nervous system such as spinal cord injury. Chondroitinase ABC is an enzyme, which promotes neurite outgrowth and regeneration. It has shown considerable promise as a therapy for these conditions. The aim of the study is to determine if targeting chondroitinase ABC expression to the neuronal axon can further enhance its ability to promote axon outgrowth. Long-distance axon regeneration has not yet been achieved, and would be a significant step in attaining functional recovery following spinal cord injury.

Methodology/Principal findings

To investigate this, neuronal cultures were transfected with constructs encoding axon-targeted chondroitinase, non-targeted chondroitinase or GFP, and the effects on neuron outgrowth and sprouting determined on substrates either permissive or inhibitory to neuron regeneration. The mechanisms underlying the observed effects were also explored. Targeting chondroitinase to the neuronal axon markedly enhances its ability to promote neurite outgrowth. The increase in neurite length is associated with an upregulation of β-integrin staining at the axonal cell surface. Staining for phosphofocal adhesion kinase, is also increased, indicating that the β-integrins are in an activated state. Expression of chondroitinase within the neurons also resulted in a decrease in expression of PTEN and RhoA, molecules which present a block to neurite outgrowth, thus identifying two of the pathways by which ChABC promotes neurite outgrowth.

Conclusions / Significance

The novel finding that targeting ChABC to the axon significantly enhances its ability to promote neurite extension, suggests that this may be an effective way of promoting long-distance axon regeneration following spinal cord injury. It could also potentially improve its efficacy in the treatment of other pathologies, where it has been shown to promote recovery, such as myocardial infarction, stroke and Parkinson’s disease.

Klíčová slova:

Axons – Cell staining – Integrins – Neurites – Neuronal differentiation – Neurons – Spinal cord injury – Transfection


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