Detection of Viral Proteins in Rabies Virus-Infected Mouse Brain Tissue

Collect fixed brain sections from intracerebral rabies virus-infected mice at defined postmortem intervals. 

The virus infects pyramidal neurons in the cerebral cortex, and expressed viral proteins localize within intracytoplasmic granular structures known as inclusion bodies.

Treat the sections with ammonium chloride to neutralize fixation-induced reactive groups and minimize background staining.

Immerse the sections in hydrogen peroxide to inactivate endogenous peroxidase and reduce nonspecific signals.

Apply a blocking solution containing a detergent to permeabilize cellular membranes and mask nonspecific binding sites.

Incubate with primary antibodies specific to the viral proteins.

Wash and incubate with biotin-conjugated secondary antibodies that bind to the primary antibodies.

Wash again and introduce peroxidase enzyme-conjugated avidin, which binds to biotin, followed by a chromogenic substrate that the enzyme converts into a colored product.

Visualize the immunoreactive inclusion bodies containing viral proteins using a microscope.

With an increase in postmortem time, neuronal degradation and the disappearance of inclusion bodies become evident.