Chronic exposure to oestrogens is known to inhibit the secretion of luteinising hormone (LH) in rats, leading to anovulation. Hypothalamic catecholamines, norepinephrine and dopamine play an important role in LH regulation. However, the effects of chronic exposure to low levels of oestradiol on hypothalamic catecholamines have not been investigated thoroughly. In the present study, adult female Sprague-Dawley rats were either sham implanted or implanted with 17β-oestradiol (E2) pellets (20 ng/day) for 30 (E-30), 60 (E-60) or 90 (E-90) days. E2 exposure affected oestrous cyclicity and ovarian morphology in a duration-dependent manner. There was no change in oestrous cyclicity in E-30 rats; however, 75% of E-60 and 95% of E-90 rats were acyclic (P < 0.05). Cycling rats from E-30 or the control group were killed at different time points on the afternoon of pro-oestrous. E-30 rats in oestrous, constant oestrous rats in the E-60 and E-90 groups and a group of old constant oestrous (OCE) rats were killed at 12.00 h. LH was measured in the serum by radioimmunoassay. Individual hypothalamic nuclei that are involved in LH regulation were microdissected and analysed for norepinephrine and dopamine levels using high-performance liquid chromatography/electrochemical detection. Norepinephrine levels in the hypothalamic nuclei increased significantly in control and E-30 groups during the afternoon of pro-oestrous, which was accompanied by a rise in LH levels (P < 0.05). On the day of oestrous, norepinephrine concentrations in hypothalamic nuclei and serum LH were significantly lower in E-60, E-90 and OCE rats compared to E-30 and control rats. On the other hand, dopamine levels declined significantly in one hypothalamic nucleus. These results indicate that chronic E2 exposure affects hypothalamic catecholamine and serum LH levels in a duration-dependent manner. This coincides well with the loss of cyclicity observed in these animals. These results suggest that repeated exposure to endogenous oestrogens could play a role in reproductive senescence.