Ovulation requires neural circuits in the brain to be sequentially exposed to estradiol and progesterone in the female rat. Estradiol-induced neuroprogesterone is essential for the luteinizing hormone (LH) surge and subsequently, ovulation to occur. The LH surge is regulated by gonadotropin-releasing hormone (GnRH) neurons in the diagonal band of Broca (DBB) which do not express progesterone receptors (PR), but are closely associated with anteroventral periventricular nucleus (AVPV) PR-expressing kisspeptin neurons. I tested the hypothesis that estradiol-induced neuroprogesterone activates AVPV kisspeptin neurons to trigger the LH surge. Inhibiting progesterone synthesis blocked estradiol induction of the LH surge that was rescued by subsequent treatment with either progesterone or DBB kisspeptin infusion. Estradiol treatment triggered a robust LH surge that was blocked by AVPV kisspeptin asODN infusion. These results support the hypothesis that neuroprogesterone induces kisspeptin release from AVPV neurons to activate DBB GnRH neurons and trigger the LH surge.