To determine whether pulsatile gonadotropin secretion occurs in the fetus, plasma immunoreactive ovine LH (oLH) concentrations were determined by homologous RIA in serial samples obtained in 51 chronically catheterized fetuses between 79 and 140 days gestation. Pulsatile secretion of LH was detected in 39 fetuses, with a peak amplitude ranging from 1.2-11.5 ng/ml (mean +/- SD, 4.5 +/- 2.3 ng/ml). The mean plasma oLH concentration between pulses was 0.4 +/- 0.23 ng/ml. The mean amplitudes of the LH pulses were similar in the age range studied, and a sex difference was not detected. The estimated interpulse interval (determined by dividing the total number of hours during which blood samples were obtained by the number of pulses demonstrated) was 2.4 h between 91 and 110 days gestation. The pattern of plasma oFSH concentration was examined in 20 fetuses between 98 and 123 days gestation. Six fetuses with gestational ages between 106 and 115 days had FSH pulses that were equal or more than 2 times the sensitivity of the RIA. This study demonstrates a pulsatile mode of LH secretion by the ovine fetal pituitary as early as 81 days gestation and provides indirect evidence for pulsatile LRF secretion by the fetal hypothalamus and an operative LRF pulse generator by midgestation, the earliest stage in gestation studied.