Cytopathic Effects in Mouse Neuroblastoma Cells during a Non-permissive Infection with a Mutant of Vesicular Stomatitis Virus

Abstract

SUMMARYMorphological changes were extensive following infection of murine neuroblastoma N-18 cells with a temperature-sensitive (ts) mutant of vesicular stomatitis virus (VSV), G31 (complementation group III), and incubation at 39 °C, a non-permissive condition for virion maturation. Incubation for 24 h after infection resulted in extensive morphological degeneration of mitochondria with over 80 % of the mitochondria having degenerated. Mitochondrial function, as determined by Janus green B supravital staining, was reduced by 81% from that in uninfected cells. Cellular ATP levels were reduced by 50% 12 h after infection. Mitochondrial degeneration still occurred in infected cells after the inactivation of lysosomes with chloroquine. Extensive cell fusion and cytoplasmic vacuole formation also occurred during the non-permissive infection with ts G31. Loss of plasma membrane integrity was not the cause of vacuole formation since 90 % of the cells were able to exclude trypan blue 24 h after infection, nor were the vacuoles the result of inactivation of the mitochondria since cyanide-poisoned cells did not form vacuoles. The cytopathic alterations observed in N-18 cells during the non-permissive infection of N-18 cells with ts G31 did not occur during the non-permissive infection of N-18 cells with ts Gll (I), ts G41 (IV), or u.v.-inactivated ts G31. However, the non-permissive infection with ts O45(V) led to mitochondrial degeneration and cytoplasmic vacuole formation, but no cell fusion occurred. These results are discussed in light of the ultrastructural features previously observed in the central nervous system of mice infected with ts G31 and cells in culture infected with wild-type VSV.

The Nucleocapsid Protein of Vesicular Stomatitis Virus Isolated from the Brains of Nude Mice is Responsible for Abated Viral RNA Synthesis at the Normal Body Temperature of Mice

“…This fusion of cells resembled the status of spongiosus described in kuru patients and in other slow virus diseases (Gibbs et al, 1968), and AIDS patients have been shown to have multinucleated cells in their CNS (Navia et al, 1986). Cultured neuroblastoma cells fused when they were infected with tsG31 VSV and incubated at a non-permissive temperature for virus replication (Hughes et al, 1979a;Dille et al, 1981).…”

Relative Neurotropism of a Recombinant Rhabdovirus Expressing a Green Fluorescent Envelope Glycoprotein

“…The same may be true for different strains or mutants of VSV (6,8). Previous work has suggested a permanent reduction in serotonin (20,21) after VSV infection.…”

Inhibition of Vesicular Stomatitis Virus Glycoprotein Expression by Chloroquine

“…Cell fusion has only been shown with nervous system-derived cells during a non-permissive infection (Hughes et al, 1979a;DiIle et al, 1982). The fusion requires the synthesis and expression of the VSV glycoprotein (G) on the infected cell surface membrane and is inhibited by the addition of glucosamine or tunicamycin that interfere with glycosylation (Hughes et al, 1979a, Dille et al, 1981.…”