Complementary Critical Functions of Zfy1 and Zfy2 in Mouse Spermatogenesis and Reproduction

PLoS Genet. 2017 Jan 23;13(1):e1006578. doi: 10.1371/journal.pgen.1006578. eCollection 2017 Jan.


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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Fertilization / genetics*
  • GPI-Linked Proteins / genetics
  • GPI-Linked Proteins / metabolism
  • Gene Deletion
  • Huntingtin Protein / genetics
  • Huntingtin Protein / metabolism
  • Male
  • Mice
  • Phosphoinositide Phospholipase C / genetics
  • Phosphoinositide Phospholipase C / metabolism
  • Phospholipase C delta / genetics
  • Phospholipase C delta / metabolism
  • Serine Endopeptidases / genetics
  • Serine Endopeptidases / metabolism
  • Spermatogenesis / genetics*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Y Chromosome / genetics


  • DNA-Binding Proteins
  • GPI-Linked Proteins
  • Htt protein, mouse
  • Huntingtin Protein
  • Transcription Factors
  • Zfy1 protein, mouse
  • Zfy2 protein, mouse
  • Phosphoinositide Phospholipase C
  • Phospholipase C delta
  • Plcd4 protein, mouse
  • Plcz1 protein, mouse
  • Prss21 protein, mouse
  • Serine Endopeptidases