Protein kinase C is required for the disappearance of MPF upon artificial activation in mouse eggs.
The aim of the present study was to investigate the implication of protein kinase C (PKC) in the mouse egg activation process. We used OAG (1-oleoyl-2-acetyl-sn-glycerol) as a PKC activator, calphostin C as a specific PKC inhibitor, and the calcium ionophore A23187 as a standard parthenogenetic agent. The exposure of zona-free eggs to 150 microM or 50 microM OAG for 10 min resulted in meiosis II completion in approximately 80% of instances. By contrast, at a lower concentration (25 microM), the PKC stimulator was ineffective as parthenogenetic agent. Shortly after the application of 150 microM OAG, the cytosolic Ca2+ concentration ([Ca2+]i) increased transiently in all the eggs examined, whereas after the addition of 50 microM OAG, [Ca2+]i remained unchanged for at least 20 min. During this period, the activity of M-phase promoting factor (MPF) dramatically decreased and most of the eggs entered anaphase except when the PKC was inhibited by calphostin C. Similarly, MPF inactivation and meiosis resumption were prevented in calphostin C-loaded eggs following treatment with A23187, even though the ionophore-induced Ca2+ signalling was not affected. Taken together, our results indicate that stimulation of PKC is a sufficient and necessary event to induce meiosis resumption in mouse eggs and strongly suggest that, in this species, the mechanism by which a transient calcium burst triggers MPF inactivation involves a PKC-dependent pathway.
Colonna
R
R
Department of Biomedical Science and Technology, University of L'Aquila, Italy.
Tatone
C
C
Francione
A
A
Rosati
F
F
Callaini
G
G
Corda
D
D
Di Francesco
L
L
D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D004075:Diglycerides / Q000494:pharmacology; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D007476:Ionophores / Q000494:pharmacology; D016200:Maturation-Promoting Factor / Q000378:metabolism*; D008540:Meiosis; D000090204:Mesothelin; D051379:Mice; D008856:Microscopy, Fluorescence; D009281:Naphthalenes / Q000494:pharmacology; D009865:Oocytes / Q000166:cytology* / Q000201:enzymology; D010312:Parthenogenesis / Q000187:drug effects; D011493:Protein Kinase C / Q000378:metabolism*; D008941:Spindle Apparatus / Q000187:drug effects
D004075:Diglycerides; D004791:Enzyme Inhibitors; D007476:Ionophores; C000715528:Msln protein, mouse; D009281:Naphthalenes; D000001:Calcimycin; C038237:1-oleoyl-2-acetylglycerol; D011493:Protein Kinase C; D016200:Maturation-Promoting Factor; C058819:calphostin C; D000090204:Mesothelin; D002118:Calcium