A role for protein kinase C during rat egg activation.
Upon sperm-egg interaction, an increase in intracellular calcium concentration ([Ca(2+)](i)) is observed. Several studies reported that cortical reaction (CR) can be triggered not only by a [Ca(2+)](i) rise but also by protein kinase C (PKC) activation. Because the CR is regarded as a Ca(2+)-dependent exocytotic process and because the calcium-dependent conventional PKCs (cPKC) alpha and beta II are considered as exocytosis mediators in various cell systems, we chose to study activation of the cPKC in the rat egg during in vivo fertilization and parthenogenetic activation. By using immunohistochemistry and confocal microscopy techniques, we demonstrated, for the first time, the activation of the cPKC alpha, beta I, and beta II during in vivo fertilization. All three isozymes examined presented translocation to the egg's plasma membrane as early as the sperm-binding stage. However, the kinetics of their translocation was not identical. Activation of cPKC alpha was obtained by the phorbol ester 12-O-tetradecanoyl phorbol-13-acetate (TPA) or by 1-oleoyl-2-acetylglycerol (OAG) but not by the calcium ionophore ionomycin. PKC alpha translocation was first detected 5-10 min after exposure to TPA and reached a maximum at 20 min, whereas in eggs activated by OAG, translocation of PKC alpha was observed almost immediately and reached a maximum within 5 min. These results suggest that, although [Ca(2+)](i) elevation on its own does not activate PKC alpha, it may accelerate OAG-induced PKC alpha activation. We also demonstrate a successful inhibition of the CR by a myristoylated PKC pseudosubstrate (myrPKCPsi), a specific PKC inhibitor. Our study suggests that exocytosis can be triggered independently either by a [Ca(2+)](i) rise or by PKC.
Eliyahu
Efrat
E
Department of Embryology and Teratology, Sackler Faculty of Medicine, Tel Aviv University, Ramat Aviv, Tel Aviv 69978, Israel.
Shalgi
Ruth
R
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002467:Cell Nucleus / Q000502:physiology; D003720:Densitometry; D004789:Enzyme Activation / Q000502:physiology; D020536:Enzyme Activators / Q000494:pharmacology; D005089:Exocytosis / Q000502:physiology; D005260:Female; D005455:Fluorescent Antibody Technique; D007527:Isoenzymes / Q000502:physiology; D008540:Meiosis / Q000187:drug effects / Q000502:physiology; D018613:Microscopy, Confocal; D010063:Ovum / Q000254:growth & development / Q000502:physiology*; D010312:Parthenogenesis / Q000502:physiology; D011247:Pregnancy; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D051381:Rats; D013084:Sperm-Ovum Interactions / Q000502:physiology*; D014178:Translocation, Genetic / Q000502:physiology
D020536:Enzyme Activators; D007527:Isoenzymes; D011493:Protein Kinase C; D002118:Calcium