Figure 3.

Functional Analysis of Warfarin-Treated Rat Epididymal Sperm and Male Fertility

(A–C) Body weights (A), organ-to-body weight ratios for the testes, epididymis, kidneys, and spleens (B) as well as the litter size (C) of rats from +15mg and +30mg warfarin-treated groups versus WT and VK1 supplemented control groups. (D) CASA quantification of the concentration of motile caput and cauda spermatozoa.

(E–H) The percentage of motile (E) and progressive (F) sperm and the comparison of VSL (G) and VAP (H) parameters of cauda sperm from warfarin-treated and control rats.

(I) Images of caput and cauda sperm during CASA experiments. Note the numerous granular vesicles in the contents of cauda epididymidis of the warfarin-treated rats but not in the controls nor the caput contents.

(J) Percentage of defective cauda sperm from warfarin-treated rats versus controls.

(K) H&E staining showing gross anatomy of epididymis of each group. Arrowheads indicate the detached epithelial cells in the lumen.

(L) Immunofluorescent labeling showing the reduced F-actin in the smooth muscle layer (arrows) in the basal compartment of warfarin-treated epididymis.

(M) Immunolabeling of γ-H2AX (red) in testicular sections showing normal spermatogenesis in the testes of each group. Nuclei are labeled in blue with DAPI.

Scale bars, 50 μm. Bar graph data herein and in subsequent figures show the means (±SD) of at least three rats or trials; ns: no significant difference; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.