This study investigates the mechanism of pseudorabies virus infection in a mouse model with micro-wounds. The focus is on how the virus spreads through the nervous system and induces neuroinflammation.
Take an anesthetized mouse with micro-wounds on the footpad that expose the stratum basale, the deepest layer of the epidermis.
This layer contains peripheral nerve fibers connected to the dorsal root ganglia, or DRG, near the spinal cord.
Apply an inoculum of pseudorabies virus, which targets the nervous system, to the wound and rub it with a needle shaft to enhance adsorption.
The virus binds to cell surface receptors, which allows the virus to enter and begin replication.
Viral particles spread to nearby cells and enter peripheral nerves.
The virus then travels to the DRG and spreads through the spinal cord to the brain, infecting neurons.
Infected neurons release DAMPs, activating resident microglia, which secrete cytokines and chemokines to attract macrophages and neutrophils.
These immune cells release reactive oxygen species and proteolytic enzymes that damage neurons, leading to neuroinflammation.
Topically apply 20 microliters of the appropriate titer of virus inoculum onto the abraded footpad. Carry out mock inoculations with medium only in the abraded footpads of control animals in parallel. After each application, gently rub the footpad 10 times with the shaft of a needle to facilitate absorption of the virus every 10 minutes for 30 minutes. When the abraded footpad is dry, allow the mice to recover with monitoring until sternal recumbency before placing the animals into individual cages for clinical follow-up and sampling.
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