Checkpoint control of the G2/M phase transition during the first mitotic cycle in mammalian eggs

Hum Reprod. 1999 Jun;14(6):1582-7. doi: 10.1093/humrep/14.6.1582.

Abstract

The high incidence of chromosomally abnormal human embryos is frequently assumed to be due to a lack of checkpoint controls operating during early embryogenesis. In our study we have analysed when these mechanisms first become functional. Mouse oocytes treated in late metaphase I with either of two different cyclin-dependent kinase inhibitors [butyrolactone 1 (BL1) or 6-dimethylaminopurine (6-DMAP)] form nuclei in the cytoplasm. BL1-treated eggs enter S-phase at 16-18 h post-treatment and, after completion of DNA synthesis, cleave to 2-cell stage embryos. 6-DMAP treatment results in the rapid initiation of DNA synthesis, its completion by 12 h and then arrest in the G2 phase. Thus, two different cell cycle stages can be obtained at the same time point after the initiation of treatment: G1- after BL1 and G2-staged nuclei after 6-DMAP treatment. That this approach greatly facilitates cell cycle studies has been shown by analysing checkpoint function during the first division. Whilst G2-staged eggs enter M phase within 2-3 h when 6-DMAP is washed out, the onset of M phase is delayed after their fusion to G1 (BL1) cells. Here M phase occurs only after the less advanced nucleus completes DNA replication. Our results indicate that checkpoints in mammalian eggs are functional during the first mitotic cycle.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • 4-Butyrolactone / analogs & derivatives
  • 4-Butyrolactone / pharmacology
  • Adenine / analogs & derivatives
  • Adenine / pharmacology
  • Animals
  • Cyclin-Dependent Kinases / antagonists & inhibitors
  • DNA / biosynthesis
  • Enzyme Inhibitors / pharmacology
  • Female
  • G1 Phase
  • G2 Phase*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Inbred CBA
  • Mitosis*
  • Oocytes / cytology*
  • Oocytes / metabolism
  • Zygote

Substances

  • Enzyme Inhibitors
  • N(6),N(6)-dimethyladenine
  • butyrolactone I
  • DNA
  • Cyclin-Dependent Kinases
  • Adenine
  • 4-Butyrolactone