Cytochemical study of the localization and organization of parental herpes simplex virus type I DNA during initial infection of the cell

Changes in the location and structural organization of parental herpes simplex virus type 1 (HSV-1) DNA during its migration from the extracellular space to the interior of the nucleus of the target cell were examined by in situ hybridization using an HSV-1 DNA probe, specific DNA staining, and autoradiography after infection of cells with tritium-labeled viruses. In situ hybridization was carried out on denatured DNA to reveal as much as possible of the HSV-1 sequence present at the surface of the sections, and also on non-denatured DNA which revealed the presence of single-stranded portions of parental DNA, both prior to and during its intracellular migration. The results from in situ hybridization and autoradiography demonstrated that a short interval of about 15 min separated the initial contact of the viruses with the cells from the entry of parental viral DNA into the nucleus. In transit, morphologically intact nucleoids were released into the cytoplasm, and swollen nucleoids which contained partially decondensed viral DNA became detectable in the juxtanuclear cytoplasm and the periphery of the nucleus among the cell chromatin fibers. Completely decondensed parental viral DNA fibers could not be distinguished structurally from cellular DNA, but their position could be revealed by the in situ hybridization label. The infective DNA became randomly distributed within all compartments of the nucleus except the matrix-associated clusters of interchromatin granules.