Volume 19, Issue 5, Pages (November 2016)

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Volume 19, Issue 5, Pages 593-598 (November 2016) Zika Virus Infects Neural Progenitors in the Adult Mouse Brain and Alters Proliferation  Hongda Li, Laura Saucedo-Cuevas, Jose A. Regla-Nava, Guoliang Chai, Nicholas Sheets, William Tang, Alexey V. Terskikh, Sujan Shresta, Joseph G. Gleeson  Cell Stem Cell  Volume 19, Issue 5, Pages 593-598 (November 2016) DOI: 10.1016/j.stem.2016.08.005 Copyright © 2016 Elsevier Inc. Terms and Conditions

Cell Stem Cell 2016 19, 593-598DOI: (10.1016/j.stem.2016.08.005) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 1 Systemic ZIKV Can Infect Neural Progenitor Cells in the Adult Brain (A) Schematic of stem cell niches in adult mouse brain. Neural progenitor cells (NPCs) located in the subventricular zone (SVZ) in the anterior forebrain (red) adjacent to the cerebral ventricles contribute new neurons to the olfactory bulb. NPCs in the subgranular zone (SGZ) in the dorsal forebrain (red) contribute new neurons to the hippocampal dentate gyrus. LV, lateral ventricle; OB, olfactory bulb; Hipp, hippocampus. (B and C) Adult TKO mice were infected with ZIKV and then sacrificed, their brains were serially sectioned, and they were immunostained for ZIKV envelope protein (green). Evidence of ZIKV was found in the SVZ and SGZ, with less expression elsewhere in adult brain. High-power inset below with arrows (yellow) highlights immunoreactive cells. LV, lateral ventricle. Scale bar, 100 μm. (D) Marker expression during neural stem cell (NSC) differentiation from radial glia to mature neuron. (E–J) Confocal images and orthogonal projection of SVZ and SGZ regions co-stained for GFAP, Nestin, SOX2, and DCX with ZIKV envelop protein, evidencing ZIKV in NPCs and immature neuron populations. White circle outlines: infected cells. LV, lateral ventricle; SVZ, subventricular zone; DG, dentate gyrus. Scale bar, 10 μm. Cell Stem Cell 2016 19, 593-598DOI: (10.1016/j.stem.2016.08.005) Copyright © 2016 Elsevier Inc. Terms and Conditions

Figure 2 ZIKV-Infected Adult NPCs Undergo Cell Death and Show Reduced Proliferation (A and C) Representative images of SVZ (A) or SGZ (C) region from ZIKV-infected brains showing more CASP3+ cells (arrows) compared to regions in mock-infected brains. Scale bar, 100 μm. (B and D) Quantification of the number of CASP3+ cells relative to mock-infected brains. All data represent means ± SEM, n = 3 animals for each group. Student’s t test, ∗p < 0.05, ∗∗p < 0.01. (E) Adult TKO mice were infected with ZIKV or mock, injected with EdU after 6 days, then sacrificed after 2 hr. ZIKV-infected animals show reduced incorporation of EdU in both SVZ and SGZ, indicating reduced entry into S-phase. Scale bar, 100 μm. (F) Reduced Ki67 staining in ZIKV-infected mice in both SVZ and SGZ, indicating reduced cell proliferation. Scale bar, 100 μm. (G) Reduced phospho-histone H3 (pH3) staining in ZIKV-infected mice in both SVZ and SGZ, indicating reduced mitotic cells. Scale bar, 100 μm. (H–J) Stereological quantification of the number of EdU+, Ki67+, and pH3+ cells in each mouse was performed in every sixth section of the entire brains from three pairs of ZIKV and mock-infected animals. All data represent means ± SEM, n = 3 animals for each group. Student’s t test, ∗p < 0.05, ∗∗p < 0.01. Cell Stem Cell 2016 19, 593-598DOI: (10.1016/j.stem.2016.08.005) Copyright © 2016 Elsevier Inc. Terms and Conditions