Highly polarized EGF receptor tyrosine kinase activity initiates egg activation in Xenopus.
Progesterone-matured Xenopus oocytes are arrested at second metaphase but resume meiosis following fertilization. To explore the role of tyrosine kinase activity and phosphatidylinositol turnover in this activation process, we caused oocytes to express three types of human epidermal growth factor receptor (EGF-R), which differ in their ability to stimulate these biochemical processes. Following mRNA injection we found that receptor expression was highly polarized, with most receptors located on the animal hemisphere. Occupancy of the wild-type EGF-R in progesterone-matured oocytes resulted in full egg activation as indicated by an activation potential, increased intracellular-free Ca2+ ([Ca2+]i), fertilization envelope liftoff, and cortical contraction. Fura-2 imaging showed that the wave of EGF-mediated Ca2+ release started in the animal hemisphere and progressed completely around the cell. These responses required receptor tyrosine kinase activity. Matured oocytes expressing the c'973 EGF-R, which possesses kinase activity but only weakly stimulates phosphatidylinositol turnover, responded differently to EGF addition. Cortical contraction and fertilization envelope liftoff appeared normal, but there was no activation potential. Significantly, [Ca2+]i was only slightly elevated and was topologically restricted to the regions expressing receptors. Our results suggest that some aspects of egg activation can occur through a tyrosine kinase pathway. However, phosphatidylinositol hydrolysis appears necessary for both amplification and propagation of signals generated locally by activated EGF-R.
Yim
D L
DL
Section of Molecular and Cellular Biology, University of California, Davis 95616.
Opresko
L K
LK
Wiley
H S
HS
Nuccitelli
R
R
D000818:Animals; D004815:Epidermal Growth Factor / Q000378:metabolism; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism*; D005260:Female; D019204:GTP-Binding Proteins / Q000502:physiology; D006801:Humans; D008540:Meiosis / Q000502:physiology; D008564:Membrane Potentials; D009865:Oocytes / Q000187:drug effects / Q000201:enzymology* / Q000502:physiology; D010716:Phosphatidylinositols / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D010566:Virulence Factors, Bordetella / Q000494:pharmacology; D014981:Xenopus; D014982:Xenopus laevis
D010716:Phosphatidylinositols; D012333:RNA, Messenger; D010566:Virulence Factors, Bordetella; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D019204:GTP-Binding Proteins