Human globozoospermia-related gene spata16 is required for sperm formation revealed by CRISPR/Cas9-mediated mouse models

Fujihara, Y.; Oji, A.; Larasati, T.; Kojima-Kita, K.; Ikawa, M. Human Globozoospermia-Related Gene Spata16 Is Required for Sperm Formation Revealed by CRISPR/Cas9-Mediated Mouse Models. Int. J. Mol. Sci. 2017, 18, 2208. https://doi.org/10.3390/ijms1810220

Sperm-borne phospholipase C zeta-1 ensures monospermic fertilization in mice

Sperm entry in mammalian oocytes triggers intracellular Ca2+ oscillations that initiate resumption of the meiotic cell cycle and subsequent activations. Here, we show that phospholipase C zeta 1 (PLCζ1) is the long-sought sperm-borne oocyte activation factor (SOAF). Plcz1 gene knockout (KO) mouse spermatozoa fail to induce Ca2+ changes in intracytoplasmic sperm injection (ICSI). In contrast to ICSI, Plcz1 KO spermatozoa induced atypical patterns of Ca2+ changes in normal fertilizations, and most of the fertilized oocytes ceased development at the 1-2-cell stage because of oocyte activation failure or polyspermy. We further discovered that both zona pellucida block to polyspermy (ZPBP) and plasma membrane block to polyspermy (PMBP) were delayed in oocytes fertilized with Plcz1 KO spermatozoa. With the observation that polyspermy is rare in astacin-like metalloendopeptidase (Astl) KO female oocytes that lack ZPBP, we conclude that PMPB plays more critical role than ZPBP in vivo. Finally, we obtained healthy pups from male mice carrying human infertile PLCZ1 mutation by single sperm ICSI supplemented with Plcz1 mRNA injection. These results suggest that mammalian spermatozoa have a primitive oocyte activation mechanism and that PLCζ1 is a SOAF that ensures oocyte activation steps for monospermic fertilization in mammals.Nozawa, K., Satouh, Y., Fujimoto, T. et al. Sperm-borne phospholipase C zeta-1 ensures monospermic fertilization in mice. Sci Rep 8, 1315 (2018). https://doi.org/10.1038/s41598-018-19497-

Transgenic mouse lines expressing the 3xFLAG-dCas9 protein for enChIP analysis

Fujita, T, Kitaura, F, Oji, A, et al. Transgenic mouse lines expressing the 3xFLAG‐dCas9 protein for enChIP analysis. Genes Cells. 2018; 23: 318– 325. https://doi.org/10.1111/gtc.1257

Structural and functional insights into IZUMO1 recognition by JUNO in mammalian fertilization

Kato, K., Satouh, Y., Nishimasu, H. et al. Structural and functional insights into IZUMO1 recognition by JUNO in mammalian fertilization. Nat Commun 7, 12198 (2016). https://doi.org/10.1038/ncomms1219

CRISPR/Cas9 mediated genome editing in ES cells and its application for chimeric analysis in mice

Oji, A., Noda, T., Fujihara, Y. et al. CRISPR/Cas9 mediated genome editing in ES cells and its application for chimeric analysis in mice. Sci Rep 6, 31666 (2016). https://doi.org/10.1038/srep3166

Spermatozoa lacking Fertilization Influencing Membrane Protein (FIMP) fail to fuse with oocytes in mice

Fujihara, Y., Lu, Y., Noda, T., Oji, A., Larasati, T., Kojima-Kita, K., . . . Ikawa, M. (2020). Spermatozoa lacking fertilization influencing membrane protein (FIMP) fail to fuse with oocytes in mice. Proceedings of the National Academy of Sciences of the United States of America, 117(17), 9393-9400. doi:10.1073/pnas.191706011

Identification of multiple male reproductive tractspecific proteins that regulate sperm migration through the oviduct in mice

Fujihara, Y., Noda, T., Kobayashi, K., Oji, A., Kobayashi, S., Matsumura, T., . . . Ikawa, M. (2019). Identification of multiple male reproductive tractspecific proteins that regulate sperm migration through the oviduct in mice. Proceedings of the National Academy of Sciences of the United States of America, 116(37), 18498-18506. doi:10.1073/pnas.190873611

Modification of single-nucleotide polymorphism in a fully humanized CYP3A mouse by genome editing technology

Abe, S., Kobayashi, K., Oji, A. et al. Modification of single-nucleotide polymorphism in a fully humanized CYP3A mouse by genome editing technology. Sci Rep 7, 15189 (2017). https://doi.org/10.1038/s41598-017-15033-

Mice with Calr mutations homologous to human CALR mutations only exhibit mild thrombocytosis

Shide, K., Kameda, T., Kamiunten, A. et al. Mice with Calr mutations homologous to human CALR mutations only exhibit mild thrombocytosis. Blood Cancer J. 9, 42 (2019). https://doi.org/10.1038/s41408-019-0202-