The protein YWHAE (14-3-3 epsilon) in spermatozoa is essential for male fertility

Andrology. 2021 Jan;9(1):312-328. doi: 10.1111/andr.12865. Epub 2020 Aug 5.


Background: Spermatogenesis is a complex biological process highlighted by synthesis and activation of proteins that regulate meiosis and cellular differentiation occur during spermatogenesis. 14-3-3 proteins are adaptor proteins that play critical roles in kinase signaling, especially for regulation of cell cycle and apoptosis in eukaryotic cells. There are seven isoforms of the 14-3-3 family proteins encoded by seven genes (β, ε, γ, η, θ/τ, ζ and σ). 14-3-3 isoforms have been shown to have many interacting partners in several tissues including testis.

Objective: While it is known that 14-3-3 proteins are expressed in the functions of testis and spermatozoon, the role for each of the seven isoforms is not known. In this study, we investigated the roles of 14-3-3η and 14-3-3ε isoforms in spermatogenesis.

Materials and methods: To study the in vivo function of 14-3-3η and 14-3-3ε in spermatogenesis, we generated testis-specific and global knockout mice for each of 14-3-3η and 14-3-3ε isoforms (CKO and GKO, respectively). Computer-assisted semen analysis was used to assess sperm motility, while immunohistochemical studies were conducted to check spermatogenesis.

Results: Although both 14-3-3η and 14-3-3ε isoforms were present in mouse testis, only the expression of 14-3-3ε, but not 14-3-3η, was detected in spermatozoa. Mice lacking 14-3-3η were normal and fertile while 14-3-3ε CKO and GKO males showed infertility. Low sperm count with higher abnormal spermatozoa was seen in 14-3-3ε CKO mice. The motility of 14-3-3ε knockout spermatozoa was lower than that of the control. A reduction in the phosphorylation of both glycogen synthase kinase 3 and PP1γ2 was also seen in spermatozoa from 14-3-3ε CKO mice, suggesting a specific role of 14-3-3ε in spermatogenesis, sperm motility, and fertility.

Discussion and conclusion: This is the first demonstration that of the seven 14-3-3 isoforms, 14-3-3ε is essential for normal sperm function and male fertility.

Keywords: 14-3-3; YWHA; infertility; sperm motility; spermatogenesis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • 14-3-3 Proteins / genetics
  • 14-3-3 Proteins / metabolism*
  • Adenosine Triphosphate / metabolism
  • Animals
  • Female
  • Fertility*
  • Glycogen Synthase Kinase 3 / metabolism
  • Male
  • Mice, Knockout
  • Mitochondria / metabolism
  • Protein Phosphatase 1 / metabolism
  • Sperm Motility
  • Spermatogenesis*
  • Spermatozoa / abnormalities
  • Spermatozoa / metabolism*
  • Testis / metabolism


  • 14-3-3 Proteins
  • Ywhae protein, mouse
  • Adenosine Triphosphate
  • Glycogen Synthase Kinase 3
  • Ppp1cc protein, mouse
  • Protein Phosphatase 1