Hydrogen peroxide induces Src family tyrosine kinase-dependent activation of Xenopus eggs.
Fertilization is accompanied by a rapid and transient calcium release in eggs, which is required for the onset of zygotic developmental program or 'egg activation'. Recently, it was found that Src family tyrosine kinase (SFK)-dependent phospholipase C (PLC) activity is necessary for the calcium transience in fertilized Xenopus eggs. The present study demonstrates that hydrogen peroxide (H2O2) stimulates protein-tyrosine phosphorylation in Xenopus eggs, which occurs primarily in the egg cortex of the animal hemisphere as revealed by indirect immunofluorescence study. Egg SFK was found to be upregulated by H2O2 while the SFK-specific inhibitor PP1 effectively blocked H2O2-induced tyrosine phosphorylation. As in fertilized eggs, PLCgamma, but not Shc, was tyrosine-phosphorylated in H2O2-treated eggs. H2O2 also caused inositol 1,4,5-trisphosphate (IP3) production and sustained calcium release. After limited application of H2O2, elevated SFK activity and tyrosine phosphorylation were quickly reversed. Under such conditions, eggs showed cortical contraction and dephosphorylation of p42 MAP kinase, both of which are indicative of egg activation. These egg activation events, as well as H2O2-induced IP3 production and calcium release, were sensitive to PP1 and PLC inhibitor U-73122. Together, the present study demonstrated that H2O2 can mimic, at least in part, early events of Xenopus egg activation that require an SFK-dependent PLC pathway.
Sato
K
K
Laboratory of Molecular Biology, Biosignal Research Center, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan. Kksato@kobe-u.ac.jp
Ogawa
K
K
Tokmakov
A A
AA
Iwasaki
T
T
Fukami
Y
Y
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004963:Estrenes / Q000494:pharmacology; D005306:Fertilization; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D015151:Immunoblotting; D020935:MAP Kinase Signaling System; D008954:Models, Biological; D010766:Phosphorylation; D010940:Plant Proteins / Q000378:metabolism; D011760:Pyrrolidinones / Q000494:pharmacology; D013050:Spectrometry, Fluorescence; D013997:Time Factors; D010738:Type C Phospholipases / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism; D015854:Up-Regulation; D014981:Xenopus / Q000196:embryology*; D019061:src-Family Kinases / Q000378:metabolism*
D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; D004963:Estrenes; C083693:PP1 protein, Physcomitrella patens; D010940:Plant Proteins; D011760:Pyrrolidinones; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; D014443:Tyrosine; D006861:Hydrogen Peroxide; D019061:src-Family Kinases; D010738:Type C Phospholipases