Figure 2
AU040320-deficient spermatozoa exhibit globozoospermia-like features. (A) SEM images of wild-type and AU040320-deficient spermatozoa demonstrate the rounded nuclear shape and lack of acrosome in all spermatozoa analysed (100% spermatozoa of each animal; n = 5). (B) Immunostaining of dissociated epididymal spermatozoa shows that cells from AU040320-deficient mice have rounded nuclei (DAPI, blue) instead of characteristic sickle shape, lack an acrosome (PNA, green) and tails (acetylated tubulin, red) coiled around the sperm head. (C) Mitochondrial midpiece (MitoTracker, red) in mutant spermatozoa is disarranged instead of forming the normal elongated thin structure at the base of the cell nucleus and extending with the flagellum (acetylated tubulin, green). (D) Ultrastructure of spermatozoa obtained with TEM reveals the absence of the acrosome in rounded-head mutant spermatozoa in all cases, with disarranged mitochondria and coiled tail in some instances (right panel, arrowheads). Additional examples of images from these analyses are shown in Supplementary Fig. S2. Ac, acrosome; m, mitochondria; n, nucleus. (E) Normal ciliary axoneme is detected in spermatozoa from AU040320 KO mice, with the microtubular organisation containing the 9 outer doublets and the central pair of microtubules. Scale bars: (A,D) 2 μm; (B,C) 5 μm; (E) 100 nm.
