Inositol trisphosphate-induced calcium release in the generation of calcium oscillations in bovine eggs

Abstract

Bovine eggs exhibit repetitive rises in intracellular calcium concentration ([Ca2+]i) in response to fertilization. The signaling pathways and Ca2 release mechanisms involved in their generation are not well characterized. This study examined the presence of a GTP-binding protein (G-protein) signaling pathway as well as the role of inositol 1,4,5-trisphosphate (InsP3) receptor (InsP3R)-mediated Ca2+ release and ryanodine receptor (RyR)-mediated Ca2+ release, the two Ca2+ receptors/channels most often thought to participate in the generation of [Ca2+]i oscillations, by injecting appropriate agonists and antagonists and monitoring their effects on Ca2+ release and pronucleus formation, injection of guanosine 5'-0-(3-thiotriphosphate) (GTP gamma [S]), which promotes G-protein-mediated phosphoinositide turnover, induced, at high concentrations, repetitive [Ca2+]i rises. Low concentrations of GTP gamma [S] were ineffective. Injection of inositol trisphosphorothioate (InsP3S3), a nonmetabolizable analogue of InsP3, evoked an immediate Ca2+ release followed by [Ca2R]i oscillations. The GTP gamma [S]- and InsP3S3-induced oscillations showed a rapid attenuation in amplitude and were terminated in about 30-60 min. Thimerosal, a thiol oxidizing agent, caused repetitive rises in [Ca2+]i by sensitizing Ca2+ injection-induced Ca2+ release. Injection of ryanodine, which stimulates Ca(2+)-induced Ca2+ release via the RyR, did not induce [Ca2+]i oscillations; and eggs into which it was preinjected exhibited normal [Ca2+]i oscillations in response to thimerosal. Preinjection of heparin, a competitive InsP3R antagonist, blocked in a dose-dependent manner the Ca2+ response to InsP3 and thimerosal, and its injection into fertilized oscillating eggs inhibited [Ca2+]i oscillations in all eggs.(ABSTRACT TRUNCATED AT 250 WORDS)