Spontaneous and Fertilization-Induced Ca2+ Oscillations in Mouse Immature Germinal Vesicle-Stage Oocytes1

Abstract

Mouse germinal vesicle (GV)-stage oocytes not only show Ca2+ oscillations in response to fertilization but also exhibit spontaneous Ca2+ oscillations during meiotic maturation in vitro. Spontaneous Ca2+ oscillations were entirely suppressed by microinjection of heparin (25 microM final intracellular concentration), an antagonist of inositol trisphosphate (IP3) receptor, whereas fertilization-induced Ca2+ oscillations were only partially inhibited by heparin even at a high dosage of 600 microM. Inhibition of endogenous IP3 generation by antagonizing phospholipase C using U73122 (20 microM final concentration) also failed to suppress the generation of fertilization-induced Ca2+ transients, suggesting that the two types of Ca2+ oscillations do not have the same dependence on IP3-induced Ca2+ release. In addition, spontaneous Ca2+ oscillations require the presence of intact GV whereas fertilization-induced Ca2+ oscillations are independent of the GV but require cytoplasm, since enucleation eliminated only spontaneous Ca2+ oscillations but not fertilization-induced Ca2+ oscillations. These results suggest that IP3-induced Ca2+ release is the primary mechanism responsible for spontaneous Ca2+ oscillations. Sperm-induced Ca2+ oscillations, however, may employ more complex mechanisms during fertilization.