Y. Shinmura et al., Migration of virus-infected neuronal cells in cerebral slice cultures of developing mouse brains after in vitro infection with murine cytomegalovirus, ACT NEUROP, 98(6), 1999, pp. 590-596
To investigate the effect of murine cytomegalovirus (MCMV) infection on the
developing mouse, brain in vitro, we developed an infection system using c
erebral slice cultures. Using a micromanipulator, the cerebral slices from
mouse embryos on day 18.5 of gestation were injected in the subventricular
zone with recombinant MCMV in which the lacZ gene was inserted into a late
gene, and were cultured for 7 days. Viral infection, detected by beta-galac
tosidase reaction, was developed at the injection sites of the slices. The
virus-infected spots in the slices were enhanced by adding tumor necrosis f
actor-a to the medium and inhibited by adding phosphonoacetic acid or ganci
clovir. Sections from paraffin-embedded slices were subjected to immunohist
ochemical analyses. Neuronal cells, labeled with 5-brorno-2-deoxyuridine 24
h before cutting the slices, migrated to the cerebral cortex in the slices
. Virus-infected neuronal cells expressing only the early viral antigen mig
rated to the cortex, whereas glial cells expressing the immediate early and
late antigens tended to remain at the injected sites. The neuronal migrati
on of infected cells was not observed in the cerebral slices from 7-day-old
mice and viral infection was not detected after injection in the cerebral
slices from 14- and 21-day-old mice. These results from these cerebral slic
es may reflect the infectious dynamics in vivo, and this system may provide
a useful model for analysis of disorders of brain development caused by CM
V.