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Updated: Jul 10, 2026
Vaccinia Reporter Viruses for Quantifying Viral Function at All Stages of Gene Expression
Published on: May 15, 2014
Advantages of using recombinant measles viruses expressing a fluorescent reporter gene with vibratome slice
M Ludlow1, W P Duprex, S L Cosby
1School of Biomedical Sciences, The Queen's University of Belfast, Northern Ireland, UK.
Aims:
In this study of experimental measles neuropathogenesis, the utility of enhanced green fluorescent protein (EGFP) as a sensitive indicator of measles virus (MV) cell-to-cell spread in the central nervous system (CNS) has been assessed in vibratome-cut brain slices to demonstrate the degree and mechanism of viral spread in the rodent CNS.
Methods:
Recombinant MVs expressing EGFP were visualized at different levels in 200-microm vibratome-cut brain sections from infected animals by confocal scanning laser microscopy (CSLM). Comparison was made with 7-microm microtome sections, stained for the N protein of measles by immunocytochemistry (ICC).
Results:
The recombinant viruses were readily visualized in infected brain tissue, with no loss of neuropathogenicity. No difference was found in the sites of infection when MV infection was detected through EGFP fluorescence or by ICC. MV-infected cells were detected in the cerebral cortex, olfactory bulb and tract, hippocampus, thalamus, hypothalamus, ependyma and subventricular zone. However, the 200-microm vibratome-cut sections and confocal microscopy proved excellent for demonstrating virus distribution in neurites and for in-depth analysis of the extent of tract infection in the white matter of the cerebral hemispheres such as selective infection of the internal capsule and anterior commissure.
Conclusions:
The use of self-tracing recombinant MVs, viewed in thick vibratome-cut sections by CSLM, demonstrated that in experimental MV neuropathogenesis the infection is selective and spreads predominately by neurites using defined anatomical pathways.
Insights
Enhanced green fluorescent protein (EGFP) effectively tracks measles virus (MV) spread in the rodent central nervous system (CNS). This method reveals selective, neurite-driven viral dissemination along defined anatomical pathways in experimental neuropathogenesis.
Area of Science:
- Neuroscience
- Virology
- Immunology
Background:
- Measles virus (MV) infection can cause severe neuropathogenesis.
- Understanding MV spread in the central nervous system (CNS) is crucial for developing effective treatments.
- Current methods for visualizing viral spread in the CNS have limitations.
Purpose of the Study:
- To assess the utility of enhanced green fluorescent protein (EGFP) as a sensitive indicator of MV cell-to-cell spread in the rodent CNS.
- To demonstrate the degree and mechanism of viral spread in the CNS using EGFP-expressing MVs.
- To analyze viral distribution in neurites and white matter tracts.
Main Methods:
- Recombinant MVs expressing EGFP were used in infected rodent brain tissue.
- Confocal scanning laser microscopy (CSLM) visualized EGFP fluorescence in 200-microm vibratome-cut brain sections.
- Immunocytochemistry (ICC) for the N protein of MV was used for comparison with 7-microm microtome sections.
Main Results:
- Recombinant MVs expressing EGFP were readily visualized without loss of neuropathogenicity.
- EGFP fluorescence and ICC detected MV infection in similar CNS regions, including the cerebral cortex, hippocampus, and thalamus.
- 200-microm sections and CSLM excelled at showing virus distribution in neurites and white matter tracts, such as the internal capsule and anterior commissure.
Conclusions:
- Self-tracing recombinant MVs visualized in thick vibratome sections by CSLM are effective for studying MV neuropathogenesis.
- MV infection in the CNS is selective and spreads predominantly via neurites.
- Viral spread follows defined anatomical pathways within the CNS.
