Normal ovarian development is dependent on stimulation of the gonadotropic hormones, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), as well as some regulatory factors locally produced in ovary, e.g. 17beta-estradiol (E2) and gonadotropin-releasing hormone (GnRH), mediated by their respective receptors. In order to elucidate the potential roles of LH, FSH, E2 and GnRH-I during early follicular development in prepubertal ducks, mRNA expression of LH-R, FSH-R, ER-β and GnRH-I in ovaries of 1-day-old (D1), 30-day-old (D30), 60-day-old (D60) and 90-day-old (D90) ducks was measured with semi-quantitative RT-PCR using β-actin as an internal standard. The ovary index (the ratio of ovary weight/body mass) did not change from D30 to D90, while the ovary weight and serum E2 levels rose progressively, indicating the prepubertal development of the ovary. Ovarian expression of FSH-R, LH-R, ER-β and GnRH-I mRNA changed greatly during this period. Abundance of FSH-R and ER-β mRNA went up gradually from D1 to D60, followed by a decline on D90. LH-R and GnRH-I mRNA expression increased from D1 to D90, reaching a peak at D90. These results indicate that the developing ovary is highly responsive to the regulation of FSH during the early stage, while close to the onset of sexual maturation, the ovary is likely more responsive to LH. In addition, the expression of GnRH-I and ER-β mRNA in the ovary suggest that GnRH-I and E2 are involved in the regulation of prepubertal follicular development in the ovary of ducks.