Extended Data Fig. 2: Growth and molecular phenotypes in F1 offspring from nABX-induced dysbiotic fathers.
From: Paternal microbiome perturbations impact offspring fitness
(a & b) Growth curves for F1 offspring fathered by control or nABX-treated sires, stratified for (a) male offspring (CON n = 87; nABX n = 96) and (b) female offspring (CON n = 85; nABX n = 85). Shown right is a representative higher-resolution timepoint (P15) of offspring body weights. Significance by a hierarchical (nested) two-tailed t-test that considers the number of litters (N) as degrees of freedom (CON = 25; nABX=28 litters). (c) Growth curves and P15 for all F1 offspring (male & female) combined (P3 n = 172/181 (CON/nABX), N = 26/28 fathers; P15 n = 164/180 (CON/nABX)). Significance by a hierarchical (nested) two-tailed t-test. (d) Distribution of fitted F1 offspring body weights at P15. There is a population shift leading to an increase in severe growth restriction (SGR) individuals amongst nABX-derived offspring. Body-weight categories are determined by Z-score relative to controls: < −3 = severe growth restriction (SGR); −3:−2 = growth restriction (GR); 2:3 = overweight (OW); >3 = obese (OB). (e) Representative images showing the range of F1 offspring growth phenotypes from nABX-treated sires at P17, spanning from normal to severe growth restriction (SGR). Obese individuals were not observed. (f) Additional phenotype comparisons between control and nABX-derived F1 offspring at indicated postnatal (P) days. Shown are three independent pairs, each from a different litter, with control and nABX born on same day. (g) Volcano plot showing significant DEGs (highlighted ochre for downregulated, green for upregulated) in brown adipose tissue (BAT) of offspring sired by dysbiotic fathers. The top enriched gene ontology terms for up- and down- regulated DEGs are shown above. p-value adjusted for multiple testing. (h) Heatmap showing the top 50 upregulated and top 50 downregulated genes in BAT from independent F1 offspring. Each column is an F1 individual from either a control or nABX-exposed father; circles indicate no observable phenotype whilst triangles indicate a physiological phenotype (SGR) observed in that individual. Data obtained from 3 litters per paternal treatment group. (i) Volcano plot showing significant differentially expressed genes in brain from F1 offspring. P-value adjusted for multiple testing. (j) Heatmap showing expression of the top 50 upregulated and downregulated genes in brain of F1 offspring according to paternal treatment. Each column is an F1 individual from either a control or nABX-exposed father; circles indicate no observable phenotype whilst triangles indicate a physiological phenotype (SGR) observed in that individual. Data obtained from 3 litters per paternal treatment group.