The pure inhibitor of cAMP-dependent protein kinase initiates Xenopus laevis meiotic maturation. A 4-step scheme for meiotic maturation.
The availability of the pure inhibitor of cAMP-dependent protein kinase prompted a re-examination of the inhibitor-induced meiotic maturation of Xenopus laevis oocytes. Injection of the inhibitor (1.5 microM) triggered 100% germinal vesicle breakdown faster than progesterone and slower than the maturation-promoting factor: at 0.15 microM, the inhibitor still triggered 100% meiosis, but with a much slower kinetics. In contrast, injection of 24 microM calmodulin resulted in less than 50% GVBD, and results were variable from female to female. Combined injection of inhibitor and calmodulin failed to show any synergism, which does not favour hypotheses according to which calmodulin acts by activation of cyclic nucleotide phosphodiesterase. The net effect of the inhibitor is to decrease the concentration of the free catalytic sub-unit of cAMP-dependent protein kinase, fully dissociated in the unstimulated oocyte, as shown by the absence of effect of pretreatment with cholera toxin on the inhibitor-induced maturation. After such decrease by about 1 microM, a maturation protein, Mp-P, is dephosphorylated by phosphoprotein phosphatases. Dephospho-Mp triggers the synthesis of MPF in cycloheximide-sensitive steps. Finally, MPF triggers GVBD in steps insensitive to cycloheximide. Evidence for such a 4-step scheme--fall in cAMP levels, then in C sub-unit levels, dephosphorylation of Mp leading to the synthesis of MPF and finally MPF-triggered GVBD--is presented and discussed.
D000818:Animals; D002147:Calmodulin / Q000494:pharmacology; D002772:Cholera Toxin / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005260:Female; D006133:Growth Substances / Q000494:pharmacology; D007700:Kinetics; D016200:Maturation-Promoting Factor; D008540:Meiosis / Q000187:drug effects*; D009865:Oocytes / Q000254:growth & development*; D010063:Ovum / Q000254:growth & development*; D011374:Progesterone / Q000494:pharmacology; D047428:Protein Kinase Inhibitors; D014982:Xenopus laevis
D002147:Calmodulin; D006133:Growth Substances; D047428:Protein Kinase Inhibitors; D011374:Progesterone; D002772:Cholera Toxin; D003513:Cycloheximide; D016200:Maturation-Promoting Factor