Herpes simplex viral and amplicon vector-mediated gene transfer into glia and neurons in organotypic spinal cord and dorsal root ganglion cultures.
The progression of neurodegenerative diseases and secondary consequences of spinal cord injury may be diminished by introducing transgenes to glia, spinal neurons, and/or sensory neurons. Organotypic cultures of spinal cord slices and dorsal root ganglia proved to be an excellent system in which to compare the relative neurotropism of a replication-defective recombinant herpes simplex virus and herpes virus-derived amplicon vectors. Hundreds of beta-galactosidase-expressing cells, transduced by the viral vectors, were observed in spinal cord slices 3 and 8 days postinfection. Immunostaining to identify the infected cell type indicated that oligodendrocytes were permissive for viral vector transduction of beta-galactosidase in the spinal cord slice, whereas neurons were not. Heparan sulfate proteoglycan, the initial receptor for herpes contact with cells, was highly expressed in the white matter of the spinal cord slice, but was negligible in the gray matter. In contrast to the spinal cord, many fewer cells were infected in the dorsal root ganglia (DRG) by these vectors, but a majority of infected cells were identified as sensory neurons. Heparan sulfate proteoglycan expression was abundant in the sensory fibers emanating from the DRG and also surrounded each neuron within the ganglion. Our results demonstrate HSV-induced transgene expression that is amenable to ex vivo assessment of its physiological impact.
Marsh
D R
DR
Neurodegeneration Research Group, The John P. Robarts Research Institute, London, Ontario, Canada N6A 5K8.
Dekaban
G A
GA
Tan
W
W
Strathdee
C A
CA
Weaver
L C
LC
D000818:Animals; D002453:Cell Cycle; D002465:Cell Movement; D002478:Cells, Cultured; D005727:Ganglia, Spinal / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006497:Heparan Sulfate / Q000096:biosynthesis / Q000276:immunology; D008856:Microscopy, Fluorescence; D009457:Neuroglia / Q000378:metabolism* / Q000821:virology; D009475:Neurons, Afferent / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D009836:Oligodendroglia / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism; D011994:Recombinant Proteins; D018139:Simplexvirus / Q000235:genetics*; D013116:Spinal Cord / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D013119:Spinal Cord Injuries / Q000628:therapy; D013127:Spinal Nerves / Q000166:cytology / Q000378:metabolism* / Q000821:virology; D019065:Virus Assembly; D001616:beta-Galactosidase / Q000096:biosynthesis
D006497:Heparan Sulfate; D016194:Receptors, N-Methyl-D-Aspartate; D011994:Recombinant Proteins; D001616:beta-Galactosidase