Figure 10

Manchette and acrosome formation based on α-tubulin and PNA immunofluorescence staining of testes squash preparations from WT and Dcaf17^−/− mice. Testes squash preparations from WT and Dcaf17^−/− testes from adult mice were stained using antibodies against α-tubulin (red) and lectin PNA (green) to analyze manchette and acrosome formation, respectively. Nucleus was stained with DAPI (blue). Representative fluorescence images of microtubule (red), acrosome (green) and nucleus (blue) staining and corresponding merge images of post-meiotic male germ cells at different steps of spermiogenesis are shown for both the genotypes (WT and Dcaf17^−/−) at similar stages. Different steps of sperimioenesis are depicted on the basis of nuclear and acrosome staining. WT post-meiotic germ cells show normal microtubule bundles (manchette) assembly (red), acrosome morphogenesis (green) and nuclear (blue) elongation during spermiogenesis. In the Dcaf17 mutant spermiogenic cells, the organization of microtubule bundles (red) around the nucleus (blue) is disrupted resulting in abnormal manchette formation, defective nuclear (blue) condensation and elongation, and abnormal acrosome (green) morphogenesis. Ectopic microtubules, defective acrosome and abnormal nuclear head shaping are noticeable in Dcaf17^−/− post-meiotic germ cells. Magnification 1000X. Scale bar: 10 µm. Schematic representations of microtubule bundles (red) assembly, acrosome (green) formation in relation to the nucleus (blue) shaping during spermiogenesis in wild-type and Dcaf17 KO spermatids are shown.