Figure 5.
Effect of Dnhd1 deficiency on typical multiple morphological abnormalities of the flagella phenotypes and infertility in male mice
(A) The mean number of pups per litter was nine in Dnhd1+/+ male mice, whereas all Dnhd1‒/‒ male mice were completely infertile. ∗∗∗ indicates p < 0.001. n = 3.
(B) Testis sizes were comparable between Dnhd1+/+ and Dnhd1‒/‒ male mice at 8 weeks of age.
(C) Testis weights were comparable between Dnhd1+/+ and Dnhd1‒/‒ male mice at 8 weeks of age. NS, not significant. n = 3.
(D) Sperm concentration comparison between Dnhd1+/+ and Dnhd1‒/‒ male mice (63.7 × 106/mL and 20.3 × 106/mL, respectively. ∗∗ indicates p < 0.01). n = 3.
(E) Sperm motility comparison between Dnhd1+/+ and Dnhd1‒/‒ male mice (70.5% and 28%, respectively. ∗∗ indicates p < 0.01). n = 3.
(F) Sperm progressive motility comparison between Dnhd1+/+ and Dnhd1‒/‒ male mice (40% and 4%, respectively. ∗∗∗ indicates p < 0.001). n = 3.
(G) Morphology of spermatozoa from Dnhd1+/+ and Dnhd1‒/‒ male mice. When compared with the normal morphology of spermatozoa obtained from Dnhd1+/+ male mice, Dnhd1‒/‒ male mice spermatozoa exhibited normal head morphologies, but aberrant flagellar morphologies, which were consistent with the clinical phenotypes observed in men harboring bi-allelic DNHD1 variants.
(H) Dnhd1 localization in spermatozoa from Dnhd1+/+ and Dnhd1‒/‒ male mice revealed by immunofluorescence staining. Anti-DNHD1 (red) localized at the sperm axoneme from Dnhd1+/+ male mice, whereas DNHD1 (red) staining was almost absent in the sperm flagella of Dnhd1‒/‒ male mice. Anti-a-tubulin indicated the flagella (green) and the nuclei of spermatozoa were DAPI labeled (blue). Scale bars: 5 mm.