Calcium, calcium release receptors, and meiotic resumption in bovine oocytes

Abstract

During maturation, mammalian oocytes undergo a series of changes that prepare them for fertilization. These events are regulated by kinases, most notably histone H1 and mitogen-activated protein kinase. Intracellular calcium ([Ca2+](i)) oscillations participate in oocyte signaling, and it has been postulated that they play a role in oocyte maturation. In these studies we investigated the association of Ca2+, Ca2+ channels, and activation of kinases in in vitro-maturating bovine oocytes. BAPTA-AM, a Ca2+ chelator, inhibited oocyte maturation and delayed activation of kinases, although spontaneous [Ca2+](i) rises were not observed in control oocytes loaded with fura-2, a Ca2+ indicator. The ability of the 1,4,5,- inositol trisphosphate receptor (InsP3R) to release Ca2+, monitored after the addition of thimerosal and myo-inositol 1,4,5-trisphosphate (InsP3), increased as maturation progressed. This may be associated with a similar increase, monitored by Western blotting, in the density of the type I InsP3R isoform during oocyte maturation. Injection of heparin, an InsP3R antagonist, blocked oocyte maturation and activation of kinases. The density of the ryanodine receptor, another Ca2+ channel, may be 30- to 100-fold lower than that of the InsP3R in bovine oocytes. Thus, our results demonstrate that [Ca2+](i) participates in the progression of meiosis and that the InsP3R may be responsible for the majority of Ca2+ release during maturation and fertilization.