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Rats with a 4-day oestrous cycle were given a single dose of atropine (100, 300, 500 or 700 mg/kg body wt) at 13.00 h on the days of oestrus, dioestrus 1, dioestrus 2 or pro-oestrus and were autopsied on the next expected day of oestrus. The doses of atropine (in mg/kg body wt) necessary to block ovulation during the cycle were 300 at oestrus, 100 at dioestrus 1 or 2 and 700 at pro-oestrus. A single dose of atropine (100 mg/kg) at oestrus, dioestrus 1 or dioestrus 2 was given at 09.00, 13.00, 17.00 or 21.00 h, autopsy again being performed on the next expected day of oestrus. The ability of atropine to block ovulation appeared to have a circadian rhythm, with a maximum blockade at 13.00 h on dioestrus 1 and dioestrus 2 and a minimum at 21.00 h on the same days. Hormone replacement (human chorionic gonadotrophin at oestrus, dioestrus 1 or 2, oestradiol benzoate at dioestrus 2 or progesterone at pro-oestrus) re-established normal ovulation in rats whose ovulation was blocked with atropine (100 mg/kg) on dioestrus 1 at 13.00 h. When ovulation was blocked with atropine but no hormone replacement had been given, rats ovulated 24 h after the next expected day of oestrus.

Results obtained in these experiments suggest the existence of a circadian rhythm of gonadotrophin secretion thoughout the oestrous cycle and a close relationship between that rhythm and the cholinergic system.