Figure 6.

ZIKV RNA Co-localization with Astrocytes and Microglia/Macrophages

T cells, neurons, and progenitor cells in the frontal cortex and hippocampus. ZIKV RNA was visualized with RNAscope (red) and cell markers by immunohistochemistry (brown) in the frontal cortex and hippocampus of ZIKV-infected animals (ZIKV+). GFAP was used to visualize astrocytes in tissue. Astrocytes showed significant overlap with viral RNA in the frontal cortex and hippocampus. (A and B) IBA-1 served as a marker for microglia and macrophages in tissue. Microglia and macrophages showed co-localization with viral RNA both in the frontal cortex (A) as well as the hippocampus (B). (C and D) CD3 was used to characterize T cells in tissue, which showed co-localization with viral RNA in the frontal cortex (C) and the hippocampus (D). (E and F) Neuronal nuclear protein, NeuN, served as a marker for neurons in tissue. Viral RNA showed co-localization with neurons in the frontal cortex (E), but, in the hippocampus (F), NeuN and viral RNA showed more proximity than overlap. (G and H) No overlap of staining is seen with ZIKV RNA and nestin, a marker of progenitor cells: frontal cortex (G) and hippocampus (H). (I and J) Representative images were taken at 20× magnification with a Keyence BZ-X700 microscope: frontal cortex (I) and hippocampus (J). Magnified images were taken at 60× and arrows were used to denote double-positive cells.