The release of gonadotrophin-releasing hormone (Gn-RH), in response to electrical stimulation of the mediobasal part of the hypothalamus incubated in vitro, was studied in both male and female rats. In male rats significant release of Gn-RH occurred during the 10-min experimental period only when the incubated tissue was stimulated at frequencies of 10 Hz or greater. There was no release when stimulated at 5 Hz. There was also no release of hormone when the mediobasal hypothalami were incubated in a Ca2+ free medium. The amount of Gn-RH released during a 10-min incubation period increased progressively as the frequency of stimulation was raised from 10--100 Hz. During short (4-min) incubation periods the effectiveness of each stimulus pulse for Gn-RH release also increased with the frequency of stimulation. However, when stimulated for 10 min there was no increase in hormone released per stimulus pulse when frequency of stimulation was raised above 10 Hz. The amount of Gn-RH released in response to stimulation at 50 Hz was greater in male rats than in females. For the females, there was no significant difference between the amounts of Gn-RH released at dioestrus and pro-oestrus. In both male and female rats gonadectomised 4 weeks prior to hypothalamic incubation, the response to electrical stimulation at 50 Hz was reduced when compared with intact controls. Indeed, for the females there was no longer a statistically significant increase in the amount of Gn-RH in the incubation medium after 50 Hz stimulation. Ovariectomised female rats, injected twice daily for 3.5 days with 20 microgram of oestradiol benzoate released Gn-RH in response to 50 Hz stimulation in the same amounts as intact control animals. By contrast, there was no recovery of Gn-RH release to normal levels in castrated male rats similarly treated with 1.25 mg testosterone propionate.