Analysis of the p21 ras system during development of meiotic competence in Xenopus laevis oocytes.
The ability of Xenopus oocytes to undergo insulin- or insulin-like growth factor 1-induced meiotic maturation develops during oogenesis, with cells 1.0 mm in diameter or larger responding in a size-dependent manner. Since insulin-induced oocyte maturation was shown previously to be p21 ras-dependent, experiments were performed to test whether a deficiency in the p21 ras system might account for meiotic incompetence in small oocytes (less than or equal to 0.9 mm diameter). Both small and large oocytes were found to contain comparable levels of membrane-associated p21, as determined by protein immunoblotting. Treatment of both small and large oocytes with 2 microM insulin for 2 hr increased endogenous levels of membrane-associated p21 by approximately 70%. Stimulation of microinjected p21-membrane association by insulin was observed to be both time- and concentration-dependent in large oocytes with an EC50 of 50 nM. In addition, comparable levels of GTPase activating protein were measured in extracts prepared from oocytes ranging from 0.8 to 1.3 mm in diameter. Therefore, the p21 system is apparently not limiting during oogenesis, and expression of some other cellular component must account for development of meiotic competence in Xenopus oocytes.
Davis
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Department of Biological Sciences, University of Denver, University Park, Colorado 80208.
Sadler
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D000818:Animals; D005260:Female; D020690:GTPase-Activating Proteins; D007328:Insulin / Q000494:pharmacology; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D008540:Meiosis / Q000187:drug effects / Q000235:genetics*; D009865:Oocytes / Q000254:growth & development; D009866:Oogenesis / Q000187:drug effects / Q000235:genetics*; D011506:Proteins / Q000378:metabolism; D016283:Proto-Oncogene Proteins p21(ras) / Q000235:genetics*; D014982:Xenopus laevis / Q000235:genetics* / Q000254:growth & development; D020703:ras GTPase-Activating Proteins
D020690:GTPase-Activating Proteins; D007328:Insulin; D011506:Proteins; D020703:ras GTPase-Activating Proteins; D007334:Insulin-Like Growth Factor I; D016283:Proto-Oncogene Proteins p21(ras)