Expression and possible roles of extracellular signal-related kinases 1-2 (ERK1-2) in mouse primordial germ cell development

J Reprod Dev. 2020 Oct 13;66(5):399-409. doi: 10.1262/jrd.2019-141. Epub 2020 May 16.

Abstract

In the present work, we described the expression and activity of extracellular signal-related kinases 1-2 (ERK1-2) in mouse primordial germ cells (PGCs) from 8.5-14.5 days post coitum (dpc) and investigated whether these kinases play a role in regulating the various processes of PGC development. Using immunofluorescence and immunoblotting to detect the active phosphorylated form of ERK1-2 (p-ERK1-2), we found that the kinases were present in most proliferating 8.5-10.5 dpc PGCs, low in 11.5 dpc PGCs, and progressively increasing between 12.5-14.5 dpc both in female and male PGCs. In vitro culture experiments showed that inhibiting activation of ERK1-2 with the MEK-specific inhibitor U0126 significantly reduced the growth of 8.5 dpc PGCs in culture but had little effect on 11.5-12.5 dpc PGCs. Moreover, we found that the inhibitor did not affect the adhesion of 11.5 dpc PGCs, but it significantly reduced their motility features onto a cell monolayer. Further, while the ability of female PGCs to begin meiosis was not significantly affected by U0126, their progression through meiotic prophase I was slowed down. Notably, the activity of ERK1-2 was necessary for maintaining the correct expression of oocyte-specific genes crucial for germ cells survival and the formation of primordial follicles.

Keywords: Extracellular signal-regulated kinases (ERKs); Gene expression; Meiosis; Oogenesis; Primordial germ cells.

MeSH terms

  • Animals
  • Butadienes / pharmacology
  • Cell Cycle
  • Cell Differentiation
  • Cell Movement
  • Cell Proliferation
  • DNA Primers / genetics
  • Enzyme Inhibitors / pharmacology
  • Female
  • Gene Expression Profiling
  • Germ Cells / cytology*
  • In Vitro Techniques
  • Male
  • Meiosis
  • Meiotic Prophase I
  • Mice
  • Mice, Transgenic
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Mitogen-Activated Protein Kinase 1 / biosynthesis*
  • Mitogen-Activated Protein Kinase 3 / biosynthesis*
  • Nitriles / pharmacology
  • Oocytes / metabolism
  • Oogenesis / genetics
  • Ovarian Follicle / cytology
  • Ovary / metabolism

Substances

  • Butadienes
  • DNA Primers
  • Enzyme Inhibitors
  • Nitriles
  • U 0126
  • Mapk1 protein, mouse
  • Mapk3 protein, mouse
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3