Role of guanylyl cyclase in fertilisation of sea urchin eggs

Abstract

A transient increase in cytosolic free calcium ion concentration ([Ca2+]i) (Ca2+-transient) takes place in the early stages of fertilisation of sea urchin eggs as well as in other animal eggs (Miyazakiet al., 1993). This transient increase in [Ca2+]ipropagates across the egg as a Ca2+wave, which is thought to be a necessary and sufficient event for egg activation (Whitaker & Swarm, 1993). In sea urchin eggs, the rise in [Ca2+], is caused by release of Ca2+from the endoplasmic reticulum (ER) via one or both of two pathways: (a) inositol 1,4,5-trisphosphate (IP3) and the inositol 1,4,5-trisphosphate receptor/channel (IP3R) or (b) cADP-ribose (cADPR) and/or cGMP and the ryanodine receptor/channel (RyR) (Berridge, 1993). The signalling pathways from sperm to ER of eggs are not yet fully explained. Recent evidence from two lines of experiments has excited more controversy. First, intracellular injection of SH2 domain of phospholipase Cγ, which produced IP3, completely inhibited the increase in [Ca2+]i(Carrollet al., 1999). Another series of experiments showed that nitric oxide (NO) gas was produced in sperm during their acrosome reaction and in eggs during fertilisation, and that the intracellular injection of NO synthase caused egg activation (Epel, this supplement). NO gas is expected to stimulate the production of cGMP by activating soluble guanylyl cyclase (Garthewaite, 1991). Thus, it seems that direct measurements of the second messenger candidates during sea urchin fertilisation are essential to an understanding of the calcium signalling pathway. We previously measured the IP3, cGMP and cADPR contents of sea urchin eggs, and compared the time courses of their changes with that of the [Ca2+]ichange (Kurodaet al., 1997). We now examine further the involvement of guanylyl cyclase in the Ca2+signalling pathway at fertilisation of sea urchin eggs.