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. 2024 Jun 18;15:5173. doi: 10.1038/s41467-024-49524-2

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — Digital spatial profiling (DSP) of tissue from control (n = 6) and ZikV (n = 6) animals was conducted using the Nanostring GeoMx DSP platform, after immunofluorescence staining to identify regions of brain and cell types. Samples from ZIKA 2 did not meet quality control metrics for downstream analysis. a ROIs were selected in triplicate for each brain, representing DGM (red), SWM (teal), and DWM (tan). b Box and whisker plots (box = 25th–75th percentile and median, whiskers = 5th–95th percentile) representing normalized counts for selected genes classically expressed by neurons (left), astrocytes (center), and oligodendrocytes (right), according to ROI. Data for graphs are provided in Source Data file. c Normalized gene expression (row-specific Z-score) of the top 35 differentially expressed (DE) genes identified in pair-wise comparison of samples across ROI and ZikV exposure. Samples (x-axis) and genes (y-axis) were clustered by calculating Euclidean distances using Ward.D2. Top row, color coding by ROI, as in (b). Second row, color coding by exposure: grey, control; black, ZikV. Bottom row: color coding by animal. Black outline highlights genes enriched in DWM that relate to myelination. d, e Volcano plots of DE genes comparing ZikV to control animals for ROIs representing (d) DWM and (e) DGM. Orange, significantly (FDR < 0.05) upregulated in ZikV; blue, significantly downregulated in ZikV; grey, FDR > 0.05 in DE comparison. f Network of 79 DE genes (FDR < 0.05) in either DWM or DGM clustered by gene ontology (GO, large nodes) representing axon function (GO:0030424 and GO:0007411) and myelination (GO:0042552 and GO:0043209). GO terms were selected by applying over-representation analysis (ORA) to DE genes in each cluster (Fig. S2a). Small nodes represent average log-fold change (color) for each gene in DWM (left half) and DGM (right half). Average gestational age (±SD) of ZikV-exposed vs CTL animals in DSP analysis = 150(±9) vs 156(±2) days; p = 0.14 by t test for DGM; 154(±8) vs 156(±2) days; p = 0.46 by t test for DWM. Source data are provided as a Source Data file.