The role of Src family kinases in egg activation.
The Src family kinases (SFKs) are believed to mediate some of the early events of egg activation at fertilization--intracellular Ca2+ increase and resumption of the second meiotic division (RMII). SFKs are both necessary and sufficient for triggering intracellular Ca2+ increase in eggs of sea urchin, sea star, Xenopus etc, but their role in mammalian eggs is not entirely determined. In this study we examined the involvement of SFKs in the events leading to Ca2+ increase in rat eggs and demonstrated their involvement in RMII. Microinjecting mRNAs of active forms of Fyn or c-Yes but not of c-Src, into ovulated eggs, triggered RMII without evoking Ca2+ increase. A specific SFKs inhibitor (SU6656) or dominant-negative (DN) forms of Fyn or c-Yes were unable to block Ca2+ oscillations rather, modulated them, in fertilized eggs or in parthenogenetically activated eggs. Moreover, inhibiting SFKs activity blocked RMII and decreased the level of cyclin B1 degradation. Our results imply participation of SFKs in the signal transduction pathway leading to egg activation, but not in the one leading to Ca2+ increase. We propose that SFKs act downstream to Ca2+ increase at the level of M-phase promoting factor (MPF).
Reut
Tomashov-Matar
TM
Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel-Aviv University, Ramat-Aviv 69978, Tel-Aviv, Israel.
Mattan
Levi
L
Dafna
Tchetchik
T
Ruth
Kaplan-Kraicer
KK
Ruth
Shalgi
S
D000818:Animals; D020013:Calcium Signaling / Q000187:drug effects; D019926:Cyclin B / Q000378:metabolism; D056744:Cyclin B1; D005089:Exocytosis / Q000187:drug effects; D005260:Female; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D008297:Male; D008540:Meiosis / Q000187:drug effects; D000090204:Mesothelin; D051379:Mice; D008845:Microinjections; D050505:Mutant Proteins / Q000378:metabolism; D010063:Ovum / Q000166:cytology / Q000187:drug effects / Q000201:enzymology* / Q000502:physiology*; D051966:Phospholipase C gamma / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D014176:Protein Biosynthesis / Q000187:drug effects; D011691:Puromycin / Q000494:pharmacology; D018075:RNA, Complementary / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D013094:Spermatozoa; D013449:Sulfonamides / Q000494:pharmacology; D019061:src-Family Kinases / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism*
C534699:CCNB1 protein, human; C534700:Ccnb1 protein, mouse; C534701:Ccnb1 protein, rat; D019926:Cyclin B; D056744:Cyclin B1; D007211:Indoles; C000715528:Msln protein, mouse; D050505:Mutant Proteins; D018075:RNA, Complementary; C416927:SU 6656; D013449:Sulfonamides; D011691:Puromycin; D019061:src-Family Kinases; D051966:Phospholipase C gamma; D000090204:Mesothelin