A 1-kb human CDCA8 promoter directs the spermatogonia-specific luciferase expression in adult testis

Gene. 2023 May 25:866:147350. doi: 10.1016/j.gene.2023.147350. Epub 2023 Mar 9.

Abstract

Cell division cycle associated 8 (CDCA8) is a component of the chromosomal passenger complex and plays an essential role in mitosis, meiosis, cancer growth, and undifferentiated state of embryonic stem cells. However, its expression and role in adult tissues remain largely uncharacterized. Here, we studied the CDCA8 transcription in adult tissues by generating a transgenic mouse model, in which the luciferase was driven by a 1-kb human CDCA8 promoter. Our previous study showed that this 1-kb promoter was active enough to dictate reporter expression faithfully reflecting endogenous CDCA8 expression. Two founder mice carrying the transgene were identified. In vivo imaging and luciferase assays in tissue lysates revealed that CDCA8 promoter was highly activated and drove robust luciferase expression in testes. Subsequently, immunohistochemical and immunofluorescent staining showed that in adult transgenic testes, the expression of luciferase was restricted to a subset of spermatogonia that were located along the basement membrane and positive for the expression of GFRA1, a consensus marker for early undifferentiated spermatogonia. These findings for the first time indicate that the CDCA8 was transcriptionally activated in testis and thus may play a role in adult spermatogenesis. Moreover, the 1-kb CDCA8 promoter could be used for spermatogonia-specific gene expression in vivo and the transgenic lines constructed here could also be used for recovery of spermatogonia from adult testes.

Keywords: CDCA8; In vivo imaging; Luciferase; Promoter; Spermatogonia; Transgenic model.

MeSH terms

  • Adult
  • Animals
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • Humans
  • Luciferases / metabolism
  • Male
  • Mice
  • Mice, Transgenic
  • Spermatogenesis / genetics
  • Spermatogonia* / metabolism
  • Testis* / metabolism

Substances

  • Luciferases
  • CDCA8 protein, human
  • Cell Cycle Proteins