Simultaneous measurement of intracellular nitric oxide and free calcium levels in chordate eggs demonstrates that nitric oxide has no role at fertilization

At fertilization in sea urchin, the free radical nitric oxide (NO) has rece ntly been suggested to cause the intracellular Ca2+ rise responsible for eg g activation. The authors suggested that NO could be a universal activator of eggs and the present study was set up to test this hypothesis. Intracell ular NO and Ca2+ levels were monitored simultaneously in eggs of the mouse or the urochordate ascidian Ascidiella aspersa. Eggs were either fertilized or sperm extracts microinjected. Sperm-induced Ca2+ rises were not associa ted with any global, or local, change in intracellular NO, although we were able to detect NO produced by the addition of a NO donor. Furthermore, the NO synthase inhibitor N-G-nitro-L-arginine methyl ester had no effect on s perm-induced Ca2+ release but did block completely ionomycin-induced NO syn thase activation. Therefore, we suggest that the current data provide evide nce that NO has no role in the fertilization of these two chordate eggs. (C ) 2001 Academic Press.