Study of transneuronal passage of pseudorabies virus in rat central nervous system by use of immunohistochemistry and in situ hybridization

Abstract

SUMMARY We studied the uptake and sequential transneuronal passage of pseudorabies virus (prv) in rat cns by use of a combination of immunohistochemistry and in situ hybridization. Protocols for rapid detection of prv by immunohistochemistry and in situ hybridization in rats with prv infection of the cns after intranasal instillation of a wild-type strain of prv were optimized in vitro, using porcine kidney-15 cells. Pseudorabies virus-specific hybridization signals appeared in the cytoplasm and nucleus of prv-infected porcine kidney-15 cells by postinoculation (pi) hour 6. In tissue sections of prv-infected rats, prv nucleic acids were detected in areas of the rat brain in close proximity to the areas in which prv antigens were evident. The prv was initially found in the nucleus of trigeminal ganglion neurons at pi hour 24. At pi hour 72, prv antigens were observed in the mid-brain, and 24 hours later, in the telencephalon. We also found evidence of specific progressive transsynaptic transmission of the virus, and, on the basis of that, we have constructed a map of the synaptic contacts and pathways in the brain. Therefore, combined use of immunohistochemistry and in situ hybridization was useful for characterizing the pathogenesis of prv in the cns of rats after intranasal inoculation, following a pattern that mimics prv infection of the natural host.