While the ovary produces the majority of estradiol (E2) in mature female primates, extra-ovarian sources contribute to E2 synthesis and action, including brain E2 regulating hypothalamic gonadotropin-releasing hormone (GnRH). In ovary-intact female rodent models, aromatase inhibition (AI) induces a PCOS-like hypergonadotropic hyperandrogenism due to absent E2-mediated negative feedback. In order to examine the role of extra-ovarian E2 on nonhuman primate gonadotropin regulation, the present study employs letrozole to elicit AI in adult female marmoset monkeys. Sixteen female marmosets (Callithrix jacchus) (>2yrs) were randomly assigned to ovary intact or ovariectomy (OVX) conditions and subsequently placed on a daily oral regimen of either ~200µl vehicle alone (ovary intact Control, n=3, OVX, n=3) or 1 mg/kg/day Letrozole in vehicle (ovary intact AI, n=4; OVX+AI, n=6). Blood samples were collected every 10 days and plasma chorionic gonadotropin (CG) and steroid hormone levels were determined by validated RIA and LC-MS/MS, respectively. Ovary intact AI-treated and OVX females exhibited elevated CG (p<0.01, p=0.004, respectively) compared with controls, and after 30 days, OVX+AI females exhibited a suprahypergonadotropic phenotype (p=0.004) compared to ovary intact+AI and OVX females. Androstenedione, A4 (p=0.03) and testosterone, T (p=0.05) were also elevated in ovary intact AI-treated females above all other groups. The current study thus confirms in a nonhuman primate that E2 depletion and diminished negative feedback in ovary-intact females engages hypergonadotropic hyperandrogenism. Additionally, we demonstrate that extra-ovarian estrogens, possibly neuroestrogens, contribute to female negative feedback regulation of gonadotropin release.