Comparative efficacy of expression of genes delivered to mouse sensory neurons with herpes virus vectors.
To achieve gene delivery to sensory neurons of the trigeminal ganglion, thymidine kinase-negative (TK-) herpes simplex viruses (HSV) containing the reporter gene lacZ (the gene for E. coli beta-galactosidase) downstream of viral (in vectors RH116 and tkLTRZ1) or mammalian (in vector NSE-lacZ-tk) promoters were inoculated onto mouse cornea and snout. Trigeminal ganglia were removed 4, 14, 30, and 60 days after inoculation with vectors and histochemically processed with 5-bromo-4-chloro-3 indolyl-beta-galactoside (X-Gal). With vector tkLTRZ1, large numbers of labeled neurons were observed in rostromedial and central trigeminal ganglion at 4 days after inoculation. A gradual decline in the number of labeled neurons was observed with this vector at subsequent time points. With vectors RH116 and NSE-lacZ-tk, smaller numbers of labeled neurons were seen at 4 days following inoculation than were observed with vector tkLTRZ1. No labeled neurons could be observed at 14 days after inoculation with vectors RH116 and NSE-lacZ-tk. Immunocytochemistry for E. coli beta-galactosidase and in situ hybridization to HSV latency-associated transcripts revealed labeled neurons in regions of the trigeminal ganglion similar to that observed with X-Gal staining. A comparable distribution of labeled neurons in trigeminal ganglion was also observed after application of the retrograde tracer Fluoro-Gold to mouse cornea and snout. These data provide evidence that retrogradely transported tk- herpes virus vectors can be used to deliver a functional gene to sensory neurons in vivo in an anatomically predictable fashion.
Davar
G
G
Neurology Service, Massachusetts General Hospital, Boston 02114.
Kramer
M F
MF
Garber
D
D
Roca
A L
AL
Andersen
J K
JK
Bebrin
W
W
Coen
D M
DM
Kosz-Vnenchak
M
M
Knipe
D M
DM
Breakefield
X O
XO
D000818:Animals; D005456:Fluorescent Dyes; D005697:Galactosides; D015870:Gene Expression / Q000502:physiology*; D005822:Genetic Vectors; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007211:Indoles; D008297:Male; D051379:Mice; D009475:Neurons, Afferent / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000235:genetics; D018139:Simplexvirus / Q000201:enzymology / Q000235:genetics*; D013266:Stilbamidines; D012668:Trigeminal Ganglion / Q000033:anatomy & histology / Q000201:enzymology; D001616:beta-Galactosidase / Q000276:immunology / Q000378:metabolism
C049774:2-hydroxy-4,4'-diamidinostilbene, methanesulfonate salt; D005456:Fluorescent Dyes; D005697:Galactosides; D007211:Indoles; D013266:Stilbamidines; D011505:Protein-Tyrosine Kinases; D001616:beta-Galactosidase; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside