Estrogen (E2) stimulates GnRH surges by coupling a daily neural signal to neuronal circuitries governing GnRH release. We have hypothesized that E2 promotes this coupling process by inducing expression of neuronal transcription factors, which are subsequently activated by neurotransmitter-mediated mechanisms representing the daily neural signal. These experiments tested the specific hypothesis that the progesterone receptor (PR) functions in this manner, viz. as an E2-induced factor whose activation is necessary for the stimulation of GnRH surges. Two complimentary experiments were performed to determine whether activation of hypothalamic PRs is obligatory for the stimulation of GnRH surges by E2. In the first, the effects of a PR antagonist on GnRH and LH surges were assessed in ovariectomized (OVX), E2-primed rats. Rats were OVX on diestrous day 2, treated with 30 microg estradiol benzoate or oil vehicle, sc, and then administered either oil vehicle or the type I antiprogestin, ZK98299 at 0900 h on proestrus. GnRH release rates and plasma LH levels were determined in each animal by microdialysis of median eminence and atrial blood sampling, respectively. Estrogen, but not oil vehicle, treatment evoked robust and contemporaneous GnRH and LH surges in animals that received no PR antagonist on proestrus. Additional treatment with ZK98299, however, completely blocked both GnRH and LH surges. In a second experiment, specific involvement of anteroventral periventricular (AVPV) PRs in E2-induced GnRH surges was assessed. Additional groups of OVX, E2-primed rats were fitted with intracerebroventricular cannulas, and PR antisense oligonucleotides were infused into the third ventricle adjacent to the AVPV to prevent expression of PR in this periventricular region. Animals infused with PR antisense oligos did not exhibit any LH surges, whereas surges were observed in saline-, missense-, and sense oligo-treated controls. Immunohistochemistry confirmed the effectiveness of PR antisense oligonucleotides in blocking PR expression. These findings provide direct support for the hypothesis that activation of PRs, specifically those in hypothalamic regions including the AVPV, is an obligatory event in the stimulation of GnRH surges by E2.