title: Complementary Critical Functions of Zfy1 and Zfy2 in Mouse Spermatogenesis and Reproduction.
abstract: The mammalian Y chromosome plays a critical role in spermatogenesis. However, the exact functions of each gene in the Y chromosome have not been completely elucidated, partly owing to difficulties in gene targeting analysis of the Y chromosome. Zfy was first proposed to be a sex determination factor, but its function in spermatogenesis has been recently elucidated. Nevertheless, Zfy gene targeting analysis has not been performed thus far. Here, we adopted the highly efficient CRISPR/Cas9 system to generate individual Zfy1 or Zfy2 knockout (KO) mice and Zfy1 and Zfy2 double knockout (Zfy1/2-DKO) mice. While individual Zfy1 or Zfy2-KO mice did not show any significant phenotypic alterations in fertility, Zfy1/2-DKO mice were infertile and displayed abnormal sperm morphology, fertilization failure, and early embryonic development failure. Mass spectrometric screening, followed by confirmation with western blot analysis, showed that PLCZ1, PLCD4, PRSS21, and HTT protein expression were significantly deceased in spermatozoa of Zfy1/2-DKO mice compared with those of wild-type mice. These results are consistent with the phenotypic changes seen in the double-mutant mice. Collectively, our strategy and findings revealed that Zfy1 and Zfy2 have redundant functions in spermatogenesis, facilitating a better understanding of fertilization failure and early embryonic development failure.
authors.affiliation: Comprehensive Reproductive Medicine, Regulation of Internal Environment and Reproduction, Systemic Organ Regulation, Graduate School, Tokyo Medical and Dental University (TMDU), Bunkyo, Tokyo, Japan.|Department of Systems BioMedicine, Tokyo Medical and Dental University (TMDU), Bunkyo, Tokyo, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Nakasuji
authors.affiliation: RIKEN BioResource Center, Tsukuba, Ibaraki, Japan.
authors.forename: Narumi
authors.initials: N
authors.lastname: Ogonuki
authors.affiliation: Department of Systems BioMedicine, Tokyo Medical and Dental University (TMDU), Bunkyo, Tokyo, Japan.
authors.forename: Tomoki
authors.initials: T
authors.lastname: Chiba
authors.affiliation: Department of Systems BioMedicine, Tokyo Medical and Dental University (TMDU), Bunkyo, Tokyo, Japan.
authors.forename: Tomomi
authors.initials: T
authors.lastname: Kato
authors.affiliation: Division of Rare Cancer Research, National Cancer Center Research Institute, Chuo, Tokyo, Japan.
authors.forename: Kumiko
authors.initials: K
authors.lastname: Shiozawa
authors.affiliation: Department of Reproductive Biology, National Center for Child Health and Development, Setagaya, Tokyo, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Yamatoya
authors.affiliation: Molecular Biology Division, Faculty of Pharmaceutical Sciences, Nagasaki International University, Sasebo, Nagasaki, Japan.
authors.forename: Hiromitsu
authors.initials: H
authors.lastname: Tanaka
authors.affiliation: Division of Rare Cancer Research, National Cancer Center Research Institute, Chuo, Tokyo, Japan.
authors.forename: Tadashi
authors.initials: T
authors.lastname: Kondo
authors.affiliation: Department of Reproductive Biology, National Center for Child Health and Development, Setagaya, Tokyo, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Miyado
authors.affiliation: Comprehensive Reproductive Medicine, Regulation of Internal Environment and Reproduction, Systemic Organ Regulation, Graduate School, Tokyo Medical and Dental University (TMDU), Bunkyo, Tokyo, Japan.
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Miyasaka
authors.affiliation: Comprehensive Reproductive Medicine, Regulation of Internal Environment and Reproduction, Systemic Organ Regulation, Graduate School, Tokyo Medical and Dental University (TMDU), Bunkyo, Tokyo, Japan.
authors.forename: Toshiro
authors.initials: T
authors.lastname: Kubota
authors.affiliation: RIKEN BioResource Center, Tsukuba, Ibaraki, Japan.
authors.forename: Atsuo
authors.initials: A
authors.lastname: Ogura
authors.affiliation: Department of Systems BioMedicine, Tokyo Medical and Dental University (TMDU), Bunkyo, Tokyo, Japan.|Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, United States of America.
authors.forename: Hiroshi
authors.identifier: 0000-0002-5215-8745
authors.initials: H
authors.lastname: Asahara
mesh_terms: D000818:Animals; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005306:Fertilization / Q000235:genetics*; D058851:GPI-Linked Proteins / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D000071058:Huntingtin Protein / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D054801:Phosphoinositide Phospholipase C / Q000235:genetics / Q000378:metabolism; D054803:Phospholipase C delta / Q000235:genetics / Q000378:metabolism; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism; D013091:Spermatogenesis / Q000235:genetics*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014998:Y Chromosome / Q000235:genetics
chemical_list: D004268:DNA-Binding Proteins; D058851:GPI-Linked Proteins; C506812:Htt protein, mouse; D000071058:Huntingtin Protein; D014157:Transcription Factors; C501342:Zfy1 protein, mouse; C501343:Zfy2 protein, mouse; D054801:Phosphoinositide Phospholipase C; D054803:Phospholipase C delta; C518031:Plcd4 protein, mouse; C499234:Plcz1 protein, mouse; C456340:Prss21 protein, mouse; D012697:Serine Endopeptidases