Retrograde viral transduction of cortical pyramidal neurons from the spinal cord.
OBJECTIVE
The primary motor pathway, the corticospinal tract, is a major target for spinal cord regeneration studies. One way of improving the regeneration of corticospinal axons is to introduce regeneration-associated genes into cortical motor neurons using viral vector delivery.


METHODS
We used an engineered Herpes Simplex virus (HSV1) with the EF1alpha promoter encoding either LacZ or GFP to transduce cortical neurons through retrograde transport following the injection of vector into adult rat striatum or spinal cord. After three-days to one-month post-injection, sections of brain and spinal cord were viewed with fluorescence microscopy or processed for LacZ histochemistry.


RESULTS
Many layer V motor cortical neurons were transduced following striatal injections. These were not corticospinal neurons as they were not fluorogold-labelled following tracer injection into spinal cord. Corticospinal neurons in both hemispheres were, however, transduced following direct vector injections into the dorsal column of spinal cord, yielding 250-400 transduced corticospinal neurons per animal. No non-pyramidal neurons or thalamic neurons were transduced by spinal injections.


CONCLUSIONS
Therefore, this HSV1.EF1alpha vector is highly effective for the transduction of corticospinal neurons without direct injection into the brain and could be used to introduce regeneration-relevant genes into these neurons with the aim of regenerating the corticospinal tract.
Groutsi
F
F
Biovex Ltd, London, UK.
Mason
M R J
MR
Anderson
P N
PN
Martins
S
S
Anesti
M
M
Coffin
R S
RS
Campbell
G
G
D000818:Animals; D002452:Cell Count / Q000379:methods; D002540:Cerebral Cortex / Q000166:cytology* / Q000502:physiology*; D036383:Ephrin-A2 / Q000235:genetics / Q000378:metabolism; D005260:Female; D015870:Gene Expression / Q000502:physiology; D049452:Green Fluorescent Proteins / Q000235:genetics; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000502:physiology; D008297:Male; D017966:Pyramidal Cells / Q000502:physiology*; D011712:Pyramidal Tracts / Q000502:physiology; D051381:Rats; D011917:Rats, Inbred Lew; D013116:Spinal Cord / Q000502:physiology*; D013266:Stilbamidines / Q000378:metabolism; D014161:Transduction, Genetic / Q000379:methods*; D001616:beta-Galactosidase / Q000235:genetics
C049774:2-hydroxy-4,4'-diamidinostilbene, methanesulfonate salt; D036383:Ephrin-A2; D013266:Stilbamidines; D049452:Green Fluorescent Proteins; D001616:beta-Galactosidase