Journal article
Long-term gene therapy causes transgene-specific changes in the morphology of regenerating retinal ganglion cells
PLoS ONE, Vol.7(2), e31061
2012
Abstract
Recombinant adeno-associated viral (rAAV) vectors can be used to introduce neurotrophic genes into injured CNS neurons, promoting survival and axonal regeneration. Gene therapy holds much promise for the treatment of neurotrauma and neurodegenerative diseases; however, neurotrophic factors are known to alter dendritic architecture, and thus we set out to determine whether such transgenes also change the morphology of transduced neurons. We compared changes in dendritic morphology of regenerating adult rat retinal ganglion cells (RGCs) after long-term transduction with rAAV2 encoding: (i) green fluorescent protein (GFP), or (ii) bi-cistronic vectors encoding GFP and ciliary neurotrophic factor (CNTF), brain-derived neurotrophic factor (BDNF) or growth-associated protein-43 (GAP43). To enhance regeneration, rats received an autologous peripheral nerve graft onto the cut optic nerve of each rAAV2 injected eye. After 5-8 months, RGCs with regenerated axons were retrogradely labeled with fluorogold (FG). Live retinal wholemounts were prepared and GFP positive (transduced) or GFP negative (non-transduced) RGCs injected iontophoretically with 2% lucifer yellow. Dendritic morphology was analyzed using Neurolucida software. Significant changes in dendritic architecture were found, in both transduced and non-transduced populations. Multivariate analysis revealed that transgenic BDNF increased dendritic field area whereas GAP43 increased dendritic complexity. CNTF decreased complexity but only in a subset of RGCs. Sholl analysis showed changes in dendritic branching in rAAV2-BDNF-GFP and rAAV2-CNTF-GFP groups and the proportion of FG positive RGCs with aberrant morphology tripled in these groups compared to controls. RGCs in all transgene groups displayed abnormal stratification. Thus in addition to promoting cell survival and axonal regeneration, vector-mediated expression of neurotrophic factors has measurable, gene-specific effects on the morphology of injured adult neurons. Such changes will likely alter the functional properties of neurons and may need to be considered when designing vector-based protocols for the treatment of neurotrauma and neurodegeneration.
Details
- Title
- Long-term gene therapy causes transgene-specific changes in the morphology of regenerating retinal ganglion cells
- Authors/Creators
- J. Rodger (Author/Creator) - The University of Western AustraliaE.S. Drummond (Author/Creator) - The University of Western AustraliaM. Hellström (Author/Creator) - The University of Western AustraliaD. Robertson (Author/Creator) - The University of Western AustraliaA.R. Harvey (Author/Creator) - The University of Western Australia
- Publication Details
- PLoS ONE, Vol.7(2), e31061
- Publisher
- Public Library of Science
- Identifiers
- 991005541216607891
- Copyright
- © 2012 Rodger et al
- Murdoch Affiliation
- Murdoch University
- Language
- English
- Resource Type
- Journal article
- Note
- 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 credited.
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- Citation topics
- 1 Clinical & Life Sciences
- 1.82 Gait & Posture
- 1.82.875 Spinal Cord Injury
- Web Of Science research areas
- Neurosciences
- ESI research areas
- Neuroscience & Behavior