Fig. 1: Metabolomic profiling reveals disruption in steroidogenesis in the testes of ZIKV-infected mice.

a, b Viral RNA copy numbers in both whole blood (a) and testes (b) were quantified in ZIKV-infected (days 0, 2, 4, 6, 8, or 10) mice using real-time quantitative PCR (qRT-PCR), and statistical significance was determined by comparing each infected group to the 0-day group. c PCA of metabolomics data of testes from Mock- (n  =  8) and ZIKV-infected (10 dpi) (n  =  8) mice. d Altered Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathways enriched by significantly altered metabolites (FDR  <  0.05) in the testes of ZIKV-infected (10 dpi) compared with the testes of Mock-infected mice. Stars represented pathways include Phenylalanine, tyrosine, and tryptophan biosynthesis (*1), and Alanine, aspartate, and glutamate metabolism (*2). The top 20 enriched pathways are shown. e Volcano plot indicating differential metabolites associated with steroid hormone biosynthesis in the testes of Mock- and ZIKV-infected (10 dpi) mice. Upregulated and downregulated metabolites (FDR  <  0.05 and fold change > 2) are colored in red and blue, respectively; others are colored in gray. f A schematic diagram of testosterone production in Leydig cells. g, h Testosterone levels of the serum (g) or testis homogenates (h) were analyzed in ZIKV-infected (days 0, 6, or 10) mice. i, j Progesterone levels of the serum (i) or testis homogenates (j) were analyzed in ZIKV-infected (days 0, 6, or 10) mice. k, l Computer-assisted sperm analysis [total (k) and motile (l)] on samples obtained from the cauda epididymis of ZIKV-infected (days 0, 6, or 10) mice immediately after euthanasia. m Histological analysis of the epididymis collected from Mock- or ZIKV-infected mice (10 dpi) stained with hematoxylin and eosin. Stars indicate spermatozoa in the cauda epididymis. Scale bar, 100 μm. The ocular lens was set to 10×, and the magnification values correspond to the objective lens used. n Representative light micrographs of sperms obtained from the cauda epididymis of Mock- or ZIKV-infected mice (10 dpi). Scale bar, 200 μm. The images in m, n are representative of three independent experiments. The data shown in a, b, g–l are the mean ± s.d (a, b, g–j: n = 5; k, l: n = 7); Statistical analysis of (a, b, g–l) was performed using a one-way analysis of variance (ANOVA), followed by Tukey’s multiple comparison test. Source data are provided in the Source Data file.