Expression and functional analyses of ephrin type-A receptor 2 in mouse spermatogonial stem cells†

Biol Reprod. 2020 Feb 12;102(1):220-232. doi: 10.1093/biolre/ioz156.


Spermatogonial stem cells (SSCs) undergo continuous self-renewal division in response to self-renewal factors. The present study identified ephrin type-A receptor 2 (EPHA2) on mouse SSCs and showed that supplementation of glial cell-derived neurotrophic factor (GDNF) and fibroblast growth factor 2 (FGF2), which are both SSC self-renewal factors, induced EPHA2 expression in cultured SSCs. Spermatogonial transplantation combined with magnetic-activated cell sorting or fluorescence-activated cell sorting also revealed that EPHA2 was expressed in SSCs. Additionally, ret proto-oncogene (RET) phosphorylation levels decreased following the knockdown (KD) of Epha2 expression via short hairpin ribonucleic acid (RNA). Although the present immunoprecipitation experiments did not reveal an association between RET with EPHA2, RET interacted with FGFR2. The Epha2 KD decreased the proliferation of cultured SSCs and inhibited the binding of cultured SSCs to laminin-coated plates. The Epha2 KD also significantly reduced the colonization of testis cells by spermatogonial transplantation. EPHA2 was also expressed in human GDNF family receptor alpha 1-positive spermatogonia. The present results indicate that SSCs express EPHA2 and suggest that it is a critical modifier of self-renewal signals in SSCs.

Keywords: Epha2; FGF2; GDNF; spermatogenesis; spermatogonia.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult Germline Stem Cells / cytology
  • Adult Germline Stem Cells / metabolism*
  • Animals
  • Cell Proliferation / physiology
  • Glial Cell Line-Derived Neurotrophic Factor / metabolism
  • Male
  • Mice
  • Phosphorylation
  • Proto-Oncogene Mas
  • RNA, Small Interfering
  • Receptors, Eph Family / genetics
  • Receptors, Eph Family / metabolism*
  • Spermatogonia / cytology
  • Spermatogonia / metabolism*
  • Testis / metabolism*


  • Glial Cell Line-Derived Neurotrophic Factor
  • MAS1 protein, human
  • Proto-Oncogene Mas
  • RNA, Small Interfering
  • Receptors, Eph Family