Fig. 4: Paternal dysbiosis induces F1 placental insufficiency.
From: Paternal microbiome perturbations impact offspring fitness
a, Left, heatmap showing DNA methylation levels across genomic features in sperm from control or nABX-treated males (n = 5 methylomes per condition) by whole-genome bisulfite-seq. Right, scatter plot of genome-wide DNA methylation (50 CpG tiles), with differential tiles highlighted. b, Heatmaps showing differential abundance of selected miRNA (left) and tRNA fragments (right) in pooled purified sperm from independent (n = 9) control or nABX males. c, PCA of transcriptomes from embryonic (E13.5) brain and placenta according to paternal exposure. d, PCA of placenta transcriptomes at E18.5 from independent litters. e, Volcano plot showing DEGs in E18.5 placenta sired by 6-week nABX-treated males. P value adjusted for multiple testing. f, Expression of key genes for placental development in E18.5 placenta. Bar indicates mean and whiskers are s.d. Each data point is an independent placenta (CON n = 5 placenta (3 litters); nABX n = 5 placenta (3 litters)). g, Ratio of fetal mass to placental mass at E18.5, depending on preconception paternal condition. Bar indicates mean, P value by unpaired two-tailed t-test (CON n = 82, nABX n = 53). h, Expression of biomarkers of pre-eclampsia (PE) in E18.5 placenta derived from nABX fathers. Bar indicates mean and whiskers are s.d. P value by multiple-testing corrected DESeq2. CON n = 5 placenta (3 litters); nABX n = 5 placenta (3 litters). i, Representative placenta sired by control or dysbiotic males (nABX), stained for DAPI (blue) and VE-cadherin (red) to demarcate the labyrinth zone (LZ). Quantification below shows LZ as a ratio of total area. Bars indicate mean ± 95% CI. Each data point is an independent placenta tissue section (CON n = 4 placenta (4 litters); nABX n = 6 placenta (6 litters)). JZ, junctional zone. P value by unpaired two-tailed t-test. j, Levels of placental growth factor (PLGF) protein (left), a marker of PE and the sFLT1/PLGF ratio (right), in F1 placenta depending on paternal regime. P value by two-tailed t-test. Bars indicate median. Each data point is an independent placenta (CON n = 12 placenta (7 litters); nABX n = 12 placenta (6 litters); avaABX n = 8 placenta (3 litters)). Asterisks indicate P values of 0.05 or less* and 0.0001 or less***. Scale bar, 100 μm. Prom, promoter.
