Mad2 is required for inhibiting securin and cyclin B degradation following spindle depolymerisation in meiosis I mouse oocytes

Reproduction. 2005 Dec;130(6):829-43. doi: 10.1530/rep.1.00856.

Abstract

Mad2 is a pivotal component of the spindle assembly checkpoint (SAC) which inhibits anaphase promoting complex/cyclo-some (APC/C) activity by sequestering Cdc20 thereby regulating the destruction of securin and cyclin B. During mitosis, spindle depolymerisation induces a robust Mad2-dependent arrest due to inhibition of securin and cyclin B destruction. In contrast to mitosis, the molecular details underpinning the meiosis I arrest experienced by mouse oocytes exposed to spindle depolymerisation remain incompletely characterised. Notably, the role of Mad2 and the fate of the anaphase-marker, securin, are unexplored. As shown previously, we find that spindle depolymerisation by nocodazole inhibits first polar body extrusion (PBE) and stabilises cyclin B and cyclin-dependent kinase 1 activity in mouse oocytes. Here we show that stabilisation of cyclin B in nocodazole can be sustained for several hours and is associated with stabilisation of securin. These effects are SAC-mediated as, in oocytes depleted of the majority of Mad2 by morpholino antisense, securin and cyclin B are destabilised and 15% of oocytes undergo PBE. This reflects premature APC/C activation as a mutant form of cyclin B lacking its APC/C degradation signal is stable in Mad2-depleted oocytes. Moreover, homologues do not disjoin during the prolonged meiosis I arrest (> 18 h) induced by nocodaozole indicating that a non-cleavage mechanism is insufficient on its own for resolution of arm cohesion in mammalian oocytes. In conclusion, when all kinetochores lack attachment and tension, mouse oocytes mount a robust Mad2-dependent meiosis I arrest which inhibits the destruction of securin and cyclin B.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carrier Proteins / metabolism*
  • Cell Cycle Proteins / antagonists & inhibitors
  • Cell Cycle Proteins / metabolism*
  • Cells, Cultured
  • Cyclin B / metabolism*
  • Cyclin B1
  • Female
  • Fluorescent Antibody Technique
  • Histones / metabolism
  • Immunoblotting / methods
  • Mad2 Proteins
  • Meiosis / physiology*
  • Mice
  • Mice, Inbred Strains
  • Microtubules / drug effects
  • Microtubules / metabolism
  • Myelin Basic Protein / metabolism
  • Nocodazole / pharmacology
  • Oligonucleotides, Antisense / pharmacology
  • Oocytes / cytology
  • Oocytes / metabolism*
  • Securin
  • Spindle Apparatus / metabolism*
  • Time Factors

Substances

  • Carrier Proteins
  • Ccnb1 protein, mouse
  • Cell Cycle Proteins
  • Cyclin B
  • Cyclin B1
  • Histones
  • Mad2 Proteins
  • Mad2l1 protein, mouse
  • Myelin Basic Protein
  • Oligonucleotides, Antisense
  • Securin
  • Nocodazole