Nanog suppresses the expression of vasa by directly regulating nlk1 in the early zebrafish embryo

Biochimie. 2017 Nov:142:93-101. doi: 10.1016/j.biochi.2017.07.014. Epub 2017 Jul 29.

Abstract

Nanog is a homeodomain transcription factor that is essential for maintenance of pluripotency and self-renewal of embryonic stem cells (ESCs). In the present study, we demonstrate that zebrafish Nanog (zNanog) directly binds to the promoter region of zebrafish nlk1 (znlk1) by ChIP-Seq analysis and that it up-regulates the expression of znlk1 in fibroblast-like embryonic cells of Danio rerio (ZEM-2S cells) and in zebrafish embryos at 30% epiboly both at the mRNA and protein levels. In addition, compared with control (MO-C) embryos at 30% epiboly, the mRNA and protein expression of vasa and the numbers of vasa-positive cells were increased in embryos injected with zNanog morpholino (MO-zNanog). Further, injection of znlk1 mRNA into zNanog-depleted embryos restored the expression of vasa and the number of vasa-positive cells. These data indicated that zNanog up-regulates the expression of znlk1 through directly binding to the znlk1 promoter, thereby suppressing the expression of vasa. Vasa is a marker gene for PGCs. Our results suggest that zNanog plays a role in restraint of PGC cell number through regulating the expression of znlk1 in the early embryonic development. The current results provide fundamental information to support further investigation regarding the regulatory mechanism of zNanog during the development of PGCs.

Keywords: Embryonic development; PGCs; Zebrafish; zNanog; znlk1.

MeSH terms

  • Animals
  • Base Sequence
  • DEAD-box RNA Helicases / genetics*
  • Embryo, Nonmammalian / metabolism*
  • Gene Expression Regulation, Developmental*
  • HEK293 Cells
  • Humans
  • Mitogen-Activated Protein Kinases / genetics
  • Mitogen-Activated Protein Kinases / metabolism*
  • Nanog Homeobox Protein / metabolism*
  • Promoter Regions, Genetic / genetics
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Zebrafish / embryology*
  • Zebrafish Proteins / genetics*

Substances

  • Nanog Homeobox Protein
  • RNA, Messenger
  • Zebrafish Proteins
  • Mitogen-Activated Protein Kinases
  • nemo-like kinase, zebrafish
  • ddx4 protein, zebrafish
  • DEAD-box RNA Helicases