Extended Data Fig. 12: Paternal nABX exposure induces molecular and physiological responses in forthcoming placenta.
From: Paternal microbiome perturbations impact offspring fitness
(a) Principal component analysis of placental transcriptomes from independent litters sired by control or nABX-treated fathers. (b) Volcano plot showing differentially expressed genes (DEG; highlighted ochre for downregulated, green for upregulated) in nascent (E13.5) placenta derived from control of nABX-exposed sires. p-value adjusted for multiple testing. (c) Heatmap showing consistent changes in gene expression between independent placenta from different litters, depending on the microbial status of the father. (d) E13.5 placental staining with IB4, a marker for endothelial cells to reveal fetoplacental vascularization within the labyrinth. nABX-derived placental tissue displayed abnormal vasculization (arrows) and reduced total vascularization, quantified right. (e) Immunofluorescent staining to determine the prevalence of trophoblast cells (arrows indicate absence) in E13.5 placenta, with quantification shown right. (f) Increased number and size of placental infarctions (lesions, arrow) in E18.5 placenta derived from CON or nABX males. (g) E18.5 placental staining with the IB4 endothelial cell marker demonstrating impaired fetoplacental vascularization within the labyrinth zone (arrowheads indicate normal blood vessels, arrows indicate abnormal). Multiple placenta from three independent litters were examined for each group, with each sired by independent fathers (CON = 3; nABX = 3). Data in panel d-f were each collected from placenta samples (CON n = 30 subregions of labyrinth, N = 5 placenta (3 litters); nABX n = 27 subregions of labyrinth, N = 5 placenta (3 litters)) and from each placenta 5 tissue sections covering different subregions of the labyrinth were analysed. It should be noted that these samples are independent litters from Fig. 4. Bar indicates mean with 95% C.I. Data analysis using unpaired student’s two-tailed t-test. Scale bars: 50 μm (d,e), 20 μm (f) and 500 μm, 20 μm (g). Placental vascular network and trophoblastic cell count were analysed using QUPath-V0.2.3.
