Studies on the mechanism for Cai-transients in sea urchin zygotes caused by refertilization and external application of sperm extract.

Sea urchin zygotes can be refertilized when they are deprived of the fertilization membrane and the hyaline layer. We have earlier reported that a transient increase of the intracellular Ca2+ concentration (Cai-transient) is induced in zygotes refertilized by sperm or treated with a sperm extract (spex) (M. Osawa et al., 1994, Dev. Biol. 166, 268-276). We investigated quantitative characteristics of the Cai-transient induced by sperm and spex, using a Ca2+ indicator, Indo-1. When sperm or spex was applied to zygotes, the peak value of the Cai-transient was 1.16 or 0.69 microM, respectively. Although these values were lower than the peak value of 1.95 microM measured during normal fertilization, the entire time courses of the three types of Cai-transients were similar. The Cai-transients during fertilization is known to be caused both by the IP3-induced Ca2+ release (IICR) and by a mechanism independent of IICR. The Cai-transients during refertilization and fertilization were not inhibited by an IP3 receptor inhibitor, heparin or by a G-protein inhibitor, GDPbetaS. However, heparin delayed the time courses of both Cai-transients. These results suggest that there may be two signal transduction pathways operating during refertilization, one dependent and the other independent of IICR. By contrast, both heparin and GDPbetaS inhibited the spex-induced Cai-transient. The IP3 content in spex-treated zygotes increased, and the spex-induced Cai-transient occurred even in the absence of external Ca2+. Cai-transient was not observed when spex was injected into zygotes. These data suggest that spex induces IICR in zygotes by activating certain cell surface receptors coupled to G-proteins.