We have investigated the effects of agonists and antagonists to γ-aminobutyric acid-A (GABA-A) and GABA-B receptors microinjected (1 μl) into the preoptic area (POA) on LH in ovariectomized (OVX) ewes with or without estrogen (E) treatment during the breeding season. Guide tubes were placed into the preoptic area of OVX ewes using lateral ventriculograms for localization of the target area. Doses of 10 μg of the GABA agonists muscimol (GABA-A) and baclofen (GABA-B) or the GABA antagonists bicuculline (GABA-A) and phaclofen (GABA-B) were injected into the POA of tame conscious animals. Jugular venous blood was collected at 10-min intervals for 3 h, the injection of GABA drug or vehicle was given, and samples were collected for a further 3 h. The plasma samples were assayed for LH. On completion of the experiments, the brains were sectioned to locate the site of injection. In the first year, 17 ewes were used, of which 16 had correct guide tube placement. In OVX sheep, both muscimol and bicuculline injections caused suppression of plasma LH concentrations, with a cessation of pulses in many instances. When OVX sheep were treated with 1.0cm Silastic implants (sc) of E for at least 7 days, there was a decrease in LH interpulse interval. In these sheep, muscimol and bicuculline injections had effects on plasma LH concentrations similar to those in OVX sheep. In addition, bicuculline was injected into the POA of OVX ewes treated with 2.0-cm E implants. Despite E treatment resulting in reduced plasma LH levels, due to an increase in the interpulse interval, bicuculline injection further suppressed plasma LH levels. Neither baclofen nor phaclofen injection had any effect on plasma LH secretion in either OVX ewes or OVX ewes given 1.0-cm E implants. In the second year, eight sheep were used, all of which had correct guide tube placement. These sheep were treated with 2.0-cm E implants and injected with phaclofen. Phaclofen had no effect on LH secretion. These results suggest that GnRH secretion is regulated by GA-BAergic neurons at the level of the GnRH cell bodies in the POA and that during the breeding season, this is effected by GABA-A and not GABA-B receptors. Our data do not support a role for preoptic GABA in mediating the negative feedback of E on GnRH secretion, since antagonists did not ameliorate the suppressive effects of E. On the other hand, GABA inputs to GnRH neurons (or afferents) may have some integral function in regulating the pulsatile secretion of GnRH in sheep.