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

Dev Biol. 2001 Jun 1;234(1):216-30. doi: 10.1006/dbio.2001.0252.

Abstract

At fertilization in sea urchin, the free radical nitric oxide (NO) has recently been suggested to cause the intracellular Ca(2+) rise responsible for egg activation. The authors suggested that NO could be a universal activator of eggs and the present study was set up to test this hypothesis. Intracellular NO and Ca(2+) 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 Ca(2+) rises were not associated 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 sperm-induced Ca(2+) release but did block completely ionomycin-induced NO synthase activation. Therefore, we suggest that the current data provide evidence that NO has no role in the fertilization of these two chordate eggs.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / metabolism*
  • Chordata, Nonvertebrate / physiology*
  • Enzyme Activation
  • Female
  • Fertilization / physiology*
  • Fluoresceins
  • Fura-2
  • Male
  • Mice
  • Microscopy, Fluorescence
  • Nitric Oxide / metabolism*
  • Nitric Oxide Synthase / metabolism
  • Ovum / physiology*
  • Sea Urchins
  • Swine
  • Urochordata
  • Vertebrates / physiology*

Substances

  • 4,5-diamino-2-(3,6-dihydroxy-9H-xanthen-9-yl)benzoic acid
  • Fluoresceins
  • Nitric Oxide
  • Nitric Oxide Synthase
  • Calcium
  • Fura-2