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Neuronal survival following remote adenovirus gene delivery
Nicholas M Boulis1, Danielle E Turner, Michael J Imperiale
1Section of Neurosurgery and Department of Neurology, Center for Gene Therapy, University of Michigan, Ann Arbor, USA. nboulis@umich.edu
Object:
Virus-mediated central nervous system gene delivery is a promising means of treating traumatized tissue or degenerative diseases. In the present study, the authors examined gene expression and neuronal survival in the spinal cord after sciatic nerve administration of an adenovirus vector expressing a LacZ reporter gene.
Methods:
The time course of adenovirus gene expression, DNA fragmentation, and neuronal density were quantified in rat lumbar spinal cord by staining for beta-galactosidase (beta-Gal), terminal deoxynucleotidyl transferase, and cresyl violet after microinjection of either saline or the reporter virus into rat sciatic nerve. The expression of beta-Gal following remote vector delivery peaked at 7 days and declined thereafter but was not accompanied by neuronal cell death, as measured by DNA fragmentation. No significant difference in spinal motor neuron density was detected between virus-treated and control rats at any time point examined. Although the spinal cords removed from rats treated with cyclosporine prior to adenovirus injection contained substantially more neurons staining for beta-Gal at 7 days (67% of total neurons), the decay in the number of stained neurons was not paralleled by a decline in motor neuron density.
Conclusions:
The authors conclude that remote gene expression is suppressed by a noncytolytic process.