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Begin with an anesthetized rat.
Make a midline skin incision between the cervical C2 and thoracic T2 vertebrae.
Next, incise along the white fibrous tissue midline and retract the first muscle layer.
Dissect the thin membranous tissue at the midline of the second muscle layer, then retract the striated longitudinal muscles.
Finally, incise the third muscle layer to expose the vertebral column.
Remove the lamina and pedicle overlying the left C4 to T1 vertebrae to reveal the C5 to C8 dorsal root ganglia or DRGs.
Inject a recombinant adeno-associated viral suspension into the DRGs.
The virus encodes a growth-promoting protein that aids in axonal regeneration.
Using forceps, crush the C5 to C8 dorsal roots to sever axonal connections from the DRG to the spinal cord, creating a controlled axonal injury.
The model is ready to study virus-mediated axonal regeneration.
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