Figure 3. BA-diet induces male hypofertility associated with testicular defects in Fxrα−/− males.

(a) Spermatozoa count in the heads of the epidydimis of wild-type and Fxrα−/−males exposed during 1 month to control or BA-diets (n = 10–20 per group). Control diet groups were arbitrarily fixed at 100%. (b) Relative percentage of sterile males; each male was bred with 2 C57Bl/6J females to analyze their reproductive capacity (n = 10–15 per group). (c) Relative testis weight normalized to body weight in wild-type and Fxrα−/− mice fed a control or BA-diet for 1 month (n = 10–20 per group). Control diet groups were arbitrarily fixed at 100%. (d) Representative micrograph of H&E-stained testis of Fxrα−/− male fed a BA-diet for 1 month. Arrows indicate tubes with complete loss of germ cells. Quantification of the number of completely destructured tubules per 100 tubules (n = 10–20 per group). (e) Germ cell proliferation in wild-type and Fxrα−/− mice exposed to control or BA-diets analyzed by PCNA staining. The number of PCNA-positive cells is indicated as the number of positive cells per 100 seminiferous tubules (n = 5–10 per group). (f ) Apoptosis in wild-type and Fxrα−/− mice exposed to control or BA-diets (n = 10–20 per group) analyzed by TUNEL staining. For the quantification of TUNEL analyses, the number of tubules with TUNEL-positive cells is indicated as the number of positive tubes per 100 seminiferous tubules (n = 10–20). Control-diet–treated mice were arbitrarily fixed at 100% for each genotype. (g) Relative intra-testicular testosterone levels in Wt and Fxrα−/−mice fed a control or BA-diet for 1 month (n = 7–10 per group). (h) Testicular mRNA accumulation of Tsx and Rhox5 normalized to β-actin mRNA levels in wild-type and Fxrα−/− mice fed a control or BA-diet for 1 month (n = 6–10 per group). Control diet groups were arbitrarily fixed at 100% for each genotype. In all of the panels, data are expressed as the means ± standard error of the mean. Statistical analysis: *P < 0.05.