The effect of age and melatonin on the activity of the neuroendocrine reproductive system was studied in young cyclic (3-5 months-old), and old acyclic (23-25 month-old) female rats. Pituitary responsiveness to a bolus of GnRH (50 ng per 100 g body weight) was assessed at both reproductive stages in control and melatonin-treated (150 micrograms melatonin per 100 g body weight each day for 1 month) groups. After this experiment, female rats were treated for another month to study the influence of ageing and melatonin on the reproductive axis. Plasma LH, FSH, prolactin, oestradiol and progesterone were measured. A positive LH response to GnRH was observed in both control groups (cyclic and acyclic). However, a response of greater magnitude was observed in old acyclic rats. Melatonin treatment reduced this increased response in acyclic rats and produced a pituitary responsiveness similar to that of young cyclic rats. FSH secretion was independent of GnRH administration in all groups, indicating desynchronization between LH and FSH secretion in response to GnRH in young animals and during senescence. No effect on prolactin was observed. Significantly higher LH (3009.11 +/- 1275.08 pg ml(-1); P < 0.05) and FSH concentrations (5879.28 +/- 1631.68 pg ml(-1); P < 0.01) were seen in acyclic control rats. After melatonin treatment, LH (811.11 +/- 89.71 pg ml(-1)) and FSH concentrations (2070 +/- 301.62 pg ml(-1)) decreased to amounts similar to those observed in young cyclic rats. However, plasma concentrations of oestradiol and progesterone were not reduced. In conclusion, the results of the present study indicate that, during ageing, the effect of melatonin is exerted primarily at the hypothalamo-pituitary axis rather than on the ovary. Melatonin restored the basal concentrations of pituitary hormones and pituitary responsiveness to similar values to those observed in young rats.

The effects of a transient doe-litter separation on plasma prolactin, FSH and oestradiol concentrations, as well as the effect on LH response to exogenous GnRH administered at the time of artificial insemination, were determined in nursing rabbits. The effects on fertility, and litter size after parturition, as well as litter survival after doe-litter separation, were also studied. Control does (n = 12) had free access to nursing, whereas biostimulated does (n = 12) were separated from their litters for 48 h before artificial insemination. Plasma prolactin concentrations were decreased 24 h after the doe-litter separation (P < 0.05). The response of prolactin to suckling reached 10 times the basal values measured on day 10 after parturition (P < 0.0001). Increased oestradiol concentrations were found during the 48 h after the doe-litter separation: at 0 h, before artificial insemination (P< 0. 0001), 1.0-2.0 h after artificial insemination (P < 0.001), at 2.5 h (P < 0.05), 3.0 h (P < 0.01), and at 3.5 h (P < 0.05) after artificial insemination. Exogenous GnRH administered at the time of artificial insemination caused a greater LH response in does previously separated from their litters during 48 h (P < 0.01). The transient doe-litter separation did not affect plasma FSH concentrations, fertility, litter size or litter survival. These results suggest that a transient separation of nursing does from their litters before artificial insemination results in a decrease in plasma prolactin concentrations that could promote growth of follicular waves, and high steroidogenesis activity, leading to increased oestradiol concentrations and inducing higher sensitivity of the pituitary gland to exogenous GnRH. These findings associated to the absence of suckling episodes would lead to higher LH response and, therefore, exert a major effect on fertility.