The goal of the experiments outlined in this proposal is to characterize remote gene delivery to the spinal cored and brainstem of the superoxide dismutase (SOD1) mouse, an animal model of ALS, using adeno- associated virus. We will utilize viral vectors whose gene products are marker proteins, either beta-galactosidase or green fluorescent protein (GFP). After peripheral injection of vector into either the mouse sciatic nerve or brachial plexus, we will examine the corresponding CNS gene expression. We expect remote delivery of recombinant genes to the CNS through peripheral nervous system (PNS) injection to offer a number of potential advantages over direct CNS injections, including minimally invasive delivery of viral vectors with the potential for repeated treatments and a reduced inflammatory response. We hope to quantify gross GFP and beta-glactosidase expression in this study as well as discover biases in terms of which CNS cell types express these proteins. We also hope to confirm that retrograde axonal transport, not diffusion, is responsible for spinal cord expression. We will do this by blocking microtubule function with peripheral colchicine injection and also by analyzing beta-galactosidase distribution after injecting the protein peripherally.
Our final aims i nvolve observing the effects of remote virus delivery on neuronal cell viability and determining whether later reinjection and also be analyzing beta-galactosidase distribution after injecting the protein peripherally.
Our final aim i nvolve observing the effects of remote virus delivery on neuronal cell viability and determining whether later reinjection of vector enhances viral gene expression. Data from this study will aid in elucidating the rational choice of viral vector(s) and mode(s) of delivery for optimal expression of therapeutic proteins in the central nervous system of the ALS mouse model.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Predoctoral Individual National Research Service Award (F31)
Project #
5F31NS043865-02
Application #
6620033
Study Section
NST-2 Subcommittee (NST)
Project Start
2002-04-01
Project End
2003-09-19
Budget Start
2003-04-01
Budget End
2003-09-19
Support Year
2
Fiscal Year
2003
Total Cost
$21,834
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Neurology
Type
Schools of Medicine
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109