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Initiation of Xenopus oocyte maturation by activation of the mitogen-activated protein kinase cascade
1Department of Genetics and Molecular Biology, Kyoto University, Japan.
Abstract:
Mitogen-activated protein kinase (MAPK) and MAPK kinase (MAPKK) are activated during Xenopus oocyte maturation concomitant with the activation of maturation promoting factor (MPF). We reported previously that an anti-MAPKK neutralizing antibody inhibited progesterone- or Mos- induced initiation of oocyte maturation. Here, we show that the injection of CL100 (also called MAPK phosphatase-1) into immature oocytes inhibited progesterone-induced oocyte maturation as well as MAPK activation and that injection of mRNA encoding a constitutively active MAPKK induced activation of histone H1 kinase and germinal vesicle breakdown in the absence of progesterone. Injection of recombinant STE11 protein (a yeast MAPKK kinase) also induced initiation of oocyte maturation. These data support the idea that the MAPKK/MAPK cascade plays an important role in oocyte maturation. Interestingly, injection of the active MAPKK mRNA or the STE11 protein resulted in induction and accumulation of Mos protein. Furthermore, in the presence of cycloheximide, the STE11-induced activation of MPF as well as the induction and accumulation of Mos was blocked, and the activation of MAPK was greatly reduced. The increase in Mos protein and the activation of MAPK by injecting cyclin A protein into immature oocytes were both blocked also by cycloheximide treatment. These results are consistent with an idea that there may exist a positive feedback loop consisting of Mos, the MAPKK/MAPK cascade, and MPF, which may be important for the initiation of oocyte maturation induced by progesterone.
Insights
The mitogen-activated protein kinase (MAPK) pathway is crucial for Xenopus oocyte maturation. A positive feedback loop involving Mos, MAPK, and maturation promoting factor (MPF) initiates this process.
Area of Science:
- Cellular Biology
- Developmental Biology
- Molecular Biology
Background:
- Mitogen-activated protein kinase (MAPK) and MAPK kinase (MAPKK) activate during Xenopus oocyte maturation alongside maturation promoting factor (MPF).
- Previous studies showed anti-MAPKK antibodies inhibit progesterone- or Mos-induced oocyte maturation.
Purpose of the Study:
- To investigate the role of the MAPKK/MAPK cascade in Xenopus oocyte maturation.
- To elucidate the potential positive feedback loop involving Mos, MAPKK/MAPK, and MPF.
Main Methods:
- Injection of CL100 (MAPK phosphatase-1) into immature oocytes.
- Injection of mRNA encoding constitutively active MAPKK.
- Injection of recombinant STE11 protein (yeast MAPKK kinase).
- Experiments conducted in the presence and absence of cycloheximide.
Main Results:
- CL100 injection inhibited progesterone-induced oocyte maturation and MAPK activation.
- Active MAPKK or STE11 induced oocyte maturation markers (histone H1 kinase, germinal vesicle breakdown) without progesterone.
- STE11 and active MAPKK induced Mos protein accumulation.
- Cycloheximide blocked STE11-induced MPF activation, Mos accumulation, and reduced MAPK activation.
- Cycloheximide also blocked cyclin A-induced Mos protein increase and MAPK activation.
Conclusions:
- The MAPKK/MAPK cascade is essential for Xenopus oocyte maturation.
- A positive feedback loop involving Mos, the MAPKK/MAPK cascade, and MPF is likely involved in initiating progesterone-induced oocyte maturation.