Fig. 3: Testicular responses to gut microbiota dysbiosis indicate a regulatory gut–germline axis.
From: Paternal microbiome perturbations impact offspring fitness
a, Boxplot showing testis mass to body weight ratio after 6 weeks of nABX treatment in males. Bar represents median and whiskers indicate 5th–95th percentile. P value by unpaired two-tailed t-test (CON n = 31; nABX n = 32). b, Representative haematoxylin and eosin stained histological sections of testes from control and nABX males. Seminiferous tubules from dysbiotic males show incidence of vacuoles formed by loss of epithelium and absence of mitotic (spermatogonial) compartments. Asterisks indicate abnormal tubules. c, Quantification of abnormal testis tubules in control and dysbiotic males. P value by nested Mann–Whitney test (CON n = 54 sections, nested into N = 4 males; nABX n = 72, N = 5; P = 0.032). Bar represents median and whiskers indicate 5th–95th percentile. d, Quantification of epithelial thickness of seminiferous tubules. P value by nested t-test (CON n = 854 tubules, N = 4 males; nABX n = 1,061, N = 4; P = 0.016). Bar represents median and whiskers indicate 5th–95th percentile. e, PCA of untargeted metabolomics profiles from independent testis of control or dysbiotic males (6 wk, 6 wk + 4 rec) and after gut microbiome recovery (6 wk + 8 rec). f, Volcano plot highlighting differentially abundant metabolites in testes after 6 weeks of nABX and during recovery to restore the gut microbiome. P value by two-tailed t-test adjusted for multiple testing. g, MA ((M (log2 ratio) and A (mean average)) plot showing gene expression changes in testes from independent (n = 5) nABX males. h, Quantitation of leptin hormone level in testes (left) and circulating plasma (right) of dysbiotic males after 6 weeks of nABX, by ELISA. Bar indicates mean with 95% CI whiskers. P value by unpaired two-tailed t-test (CON n = 9; nABX n = 9). i, Testes features in males that are leptin deficient (ob/ob) for 6 weeks and wild-type controls (WT n = 3; ob/ob n = 3). Left, haematoxylin and eosin stained tubule sections; stars indicate abnormal histo-architecture. Right, testis weight, P value by unpaired two-tailed t-test and bar indicates mean with 95% CI. j, Transcriptome PCA of blastocysts derived from leptin-deficient or control fathers. Data points represent single-embryos from duplicate independent IVF experiments, each with triplicate independent fathers and littermate fathers as controls. Scale bars, 100 μm (left panel) or 50 μm (right panel) (b); 50 μm (i). FC, fold change.
