Acute and chronic stress-like levels of cortisol inhibit the oestradiol stimulus to induce sexual receptivity but have no effect on sexual attractivity or proceptivity in female sheep.

Stress-like levels of cortisol inhibit sexual receptivity in ewes but the mechanism of this action is not understood. One possibility is that cortisol interferes with the actions of oestradiol to induce sexual receptivity. We tested this hypothesis in 2 experiments with ovariectomised ewes that were artificially induced into oestrus by 12 days of i.m. injections of progesterone followed by an i.m. injection of oestradiol benzoate (ODB) 48 h later. In Experiment 1, ewes were randomly allocated to the following groups: saline infusion+25 µg ODB, saline infusion+50 µg ODB, cortisol infusion+25 µg ODB or cortisol infusion+50 µg ODB (n=5 per group). Saline or cortisol was infused i.v. for 40 h beginning at the ODB injection. In Experiment 2, ewes were infused with saline or cortisol (n=5 per group) for 5h beginning 1h before ODB injection. In both experiments, ewe sexual behaviour (attractivity, proceptivity and receptivity) was quantified every 6h. Blood samples were also collected. The cortisol infusion yielded plasma concentrations of cortisol similar to those seen during psychosocial stress. In both experiments, cortisol suppressed receptivity index (number of immobilisations by ewe/courtship displays by ram) and the number of times ewes were mounted but had no effect on attractivity or proceptivity, irrespective of the dose of ODB (Experiment 1). Cortisol also suppressed LH pulse amplitude. These results suggest that both an acute (5h) and chronic (40 h) infusion of cortisol inhibit oestradiol-induced sexual receptivity in ewes and that increasing the dose of ODB does not overcome the inhibitory effects of cortisol.

Evidence that RF-amide related peptide-3 is not a mediator of the inhibitory effects of psychosocial stress on gonadotrophin secretion in ovariectomised ewes.

It is well known that stress inhibits normal reproductive function, including gonadotrophin secretion; however, the mechanisms and mediators involved are largely unknown. Stress impairs the secretion of luteinising hormone (LH), and it has been suggested that the RF-amide gonadotrophin-inhibitory hormone (GnIH), known as RF-amide related peptide-3 (RFRP-3) in mammalian species, may mediate this inhibitory effect of stress. If this is the case, the GnIH/RFRP system would likely be up-regulated during stress. We tested this hypothesis in ovariectomised ewes using a psychosocial stressor: isolation/restraint. Ewes were randomly allocated to control or stress (n=5 per group). Isolation/restraint stress was imposed for 90 min after control sampling for 4 h, whereas control ewes were sampled continuously for 5.5 h. All ewes were then euthanased and brains were collected. As expected, plasma concentrations of cortisol were increased significantly (P<0.05) by stress and plasma concentrations of LH were significantly (P<0.05) reduced. Immunohistochemistry and in situ hybridisation were conducted for RFRP-3 peptide and RFRP mRNA expression, respectively, in the paraventricular nucleus/dorsal medial hypothalamus region of the hypothalamus. There was no significant effect of stress on RFRP-3 peptide or mRNA levels, with no differences between control or stress ewes. Furthermore, there was no difference in the number of RFRP-3 cells double-labelled for Fos between control and stress ewes and there was no difference in the cellular expression of RFRP mRNA between groups. These results indicate that the GnIH/RFRP system is not activated by psychosocial stress in ewes, suggesting that it is an unlikely mediator of the effects of stress on LH secretion.

Psychosocial stress suppresses attractivity, proceptivity and pulsatile LH secretion in the ewe.

Various stressors suppress pulsatile secretion of luteinizing hormone (LH) in ewes and cortisol has been shown to be a mediator of this effect under various conditions. In contrast, little is known about the impact of stress and cortisol on sexual behavior in the ewe. Therefore, we tested the hypothesis that both psychosocial stress and stress-like levels of cortisol will reduce the level of attractivity, proceptivity and receptivity in addition to suppressing LH secretion in the ewe. In Experiment 1, a layered stress paradigm of psychosocial stress was used, consisting of isolation for 4 h with the addition of restraint, blindfold and noise of a barking dog (predator stress) at hourly intervals. This stress paradigm reduced LH pulse amplitude in ovariectomized ewes. In Experiment 2, ovariectomized ewes were artificially induced into estrus with progesterone and estradiol benzoate treatment and the layered stress paradigm was applied. LH was measured and sexual behavior was assessed using T-mazes and mating tests. Stress reduced pulsatile LH secretion, and also reduced attractivity and proceptivity of ewes but had no effect on receptivity. In Experiment 3, ewes artificially induced into estrus were infused with cortisol for 30 h. Cortisol elevated circulating plasma concentrations of cortisol, delayed the onset of estrus and resulted in increased circling behavior of ewes (i.e. moderate avoidance) during estrus and increased investigation and courtship from rams. There was no effect of cortisol on attractivity, proceptivity or receptivity during estrus. We conclude that psychosocial stress inhibits LH secretion, the ability of ewes to attract rams (attractivity) and the motivation of ewes to seek rams and initiate mating (proceptivity), but cortisol is unlikely to be the principal mediator of these effects.