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Frog virus 3 replication: electron microscope observations on the sequence of infection in chick embryo fibroblasts
Abstract:
The replication of frog virus 3 in primary chick embryo fibroblasts has been studied by examination of thin sections with the electron microscope and the assay of infectious viurs. Uptake of frog virus 3 by the cells was observed to occur by pinocytosis and this may be the method of entry. Early in infection (i h p.i.) marked margination of the nuclear chromatin occurred and the chromatin remained in this condition throughtout the infection. Foci of infection were first detected in the cytoplasm of cells 24 h p.i. when production of infectious virus commenced. These foci appeared as electron translucent areas containing fine grains, surrounded by degenerate mitochondria. The foci usually contained virus particles. At this time budding of virus particles at the plasma membrane occurred. Later in infection at 36 and 48 h p.i. large numbers of virus particles were detected in the cytoplasm of cells either scattered loosely throughtout the cell, arranged as clusters or in paracrystalline arrays. Extensive budding at the plasma membrane then took place. Virus particles were detected in the nucleus of the cells at these late stages and it is possible that the virus may infect and replicate at this site. Throughout the productive stages of infection aberrant forms of the virus, namely particles devoid of cores, incompletely assembled particles and elongated bacilliform particles were noticed.
Insights
Frog virus 3 enters chick embryo cells via pinocytosis, replicating in the cytoplasm and potentially the nucleus. Aberrant viral forms were observed during infection.
Area of Science:
- Virology
- Cell Biology
Background:
- Frog virus 3 (FV3) is an aquatic virus with a broad host range.
- Understanding FV3 replication is crucial for studying Ranavirus pathogenesis.
Purpose of the Study:
- To elucidate the replication cycle of frog virus 3 in primary chick embryo fibroblasts.
- To characterize the cytopathic effects and viral assembly during FV3 infection.
Main Methods:
- Thin section electron microscopy was used to visualize viral replication.
- Assays for infectious virus were performed to quantify viral production.
Main Results:
- FV3 entry into cells occurred via pinocytosis.
- Viral replication initiated in the cytoplasm at 24 hours post-infection, with foci containing virus particles and degenerate mitochondria.
- Extensive virus budding at the plasma membrane and paracrystalline arrays of virus particles were observed.
- Virus particles were detected in the nucleus at later stages, suggesting potential nuclear replication.
- Aberrant viral forms, including coreless, incomplete, and elongated particles, were noted.
Conclusions:
- FV3 replicates in chick embryo fibroblasts, with distinct cytoplasmic and potential nuclear stages.
- Pinocytosis is the likely entry mechanism for FV3.
- Aberrant viral morphogenesis occurs during FV3 replication.