The cortical granules of hamster ova were ruptured in vitro by electrical stimulation (standard treatment was 150 V, 1·0 msec). Approximately 8 min were required for the cortical granule material so released to induce a complete zona reaction. The active material from the cortical granules was released into the medium from vitelli and was shown to act directly on the zona pellucida and on mouse, as well as on hamster, ova, i.e. its action is not species specific. The action of the cortical granule material was reversed by trypsin inhibitors. Such a tryspin-like protease, causing the zona reaction, was also released from vitelli by spermatozoa.