Neural substrates mediating the negative feedback effects of oestrogen on luteinising hormone-releasing hormone (LHRH) release were studied using the in vivo push-pull perfusion method in female rhesus monkeys. Twelve long-term ovariectomized female monkeys were implanted with Silastic capsules containing 17beta-oestradiol 2 weeks before the experiments and, on the day of the experiment, oestradiol benzoate (EB, 50 microg/kg) or oil was subcutaneously injected. Push-pull perfusate samples from the stalk-median eminence were collected in 10-min fractions from 4 h before to 18-20 h after EB or oil injection. LHRH and neuropeptide Y (NPY) levels in the same perfusates were measured by radioimmunoassay, and glutamate and GABA in the same perfusates were assessed by high-performance liquid chromatography (HPLC). The results indicate that EB significantly suppressed LHRH release (P < 0.005) starting within 2 h after EB, and continued for 18 h or until the experiment was terminated. Pulse analysis suggested that oestrogen suppressed the pulse amplitude, but not pulse frequency, of LHRH release. By contrast, EB did not alter any parameters (mean release, pulse amplitude or frequency) of pulsatile NPY release throughout the experiment. HPLC analysis further suggested that neither glutamate nor GABA levels in the stalk-median eminence were changed with oestrogen-induced LHRH suppression. Oil treatment did not alter LHRH, NPY, GABA and glutamate levels. It is concluded that oestrogen induces suppression of pulsatile LHRH release within 2 h, but the inhibitory effect of oestrogen on LHRH release does not appear to be mediated by NPY, GABAergic, or glutamatergic neurones.