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Matrix‐mediated gene transfer to brain cortex and dorsal root ganglion neurones by retrograde axonal transport after dorsal column lesion
Author(s) -
Gonzalez Ana Maria,
Berry Martin,
Greenlees Lydia,
Logan Ann,
Baird Andrew
Publication year - 2006
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.919
Subject(s) - dorsal root ganglion , lesion , glial scar , biology , anatomy , neurotrophic factors , pathology , microbiology and biotechnology , spinal cord , chemistry , spinal cord injury , neuroscience , medicine , receptor , biochemistry
Abstract Background In previous studies, we showed that the immobilisation of DNAs encoding basic fibroblast growth factor, neurotrophin‐3 and brain‐derived neurotrophic factor in a gene‐activated matrix (GAM) promotes sustained survival of axotomised retinal ganglion cells after optic nerve injury. Here, we evaluated if the immobilisation of DNAs in a GAM could be an effective approach to deliver genes to axotomised dorsal root ganglion (DRG) neurones after spinal cord injury and if the matrix component of the GAM would modulate the deposition of a dense scar at the injury site. Methods We evaluated the expression of the thymidine kinase (TK) reporter gene in brain cortex and DRG after a bilateral T8 dorsal column (DC) lesion using PCR, RT‐PCR and in situ hybridisation analyses. Collagen‐based GAMs were implanted at the lesion site and the cellular response to the GAM was assessed using cell‐specific markers. Results At 1 week post‐injury, PCR analyses confirmed that DNA TK was retrogradely transported from the DC lesion where the GAM was implanted to the brain cortex and to caudal DRG neurones, and RT‐PCR analyses showed expression of mRNA TK . At 7 weeks post‐injury, DNA TK was still be detected in the GAM and DRG. In situ hybridisation localised DNA TK and mRNA TK within fibroblasts, glia, endothelial and inflammatory cells invading the GAM and in DRG neurones. Interestingly, the presence of a GAM also reduced secondary cavitation and scar deposition at the lesion site. Conclusions These results establish that GAMs act as bridging scaffolds in DC lesions limiting cavitation and scarring and delivering genes both locally to injury‐reactive cells and distally to the cerebral cortex and to DRG neuronal somata through retrograde axonal transport. Copyright © 2006 John Wiley & Sons, Ltd.