Norepinephrine (NE) and dopamine (DA) actively inhibit the release of LH in anestrous ewes. This can be detected as an increase in LH pulse frequency following i.v. injection of NE and DA antagonists. The objective of this study was to determine the sites of these inhibitory actions in the ovine hypothalamus by using local administrations of the NE antagonist, phenoxybenzamine (PBZ), or the DA antagonist, pimozide (PIM), into specific hypothalamic areas. Each neurotransmitter antagonist was administered via a chronically implanted steel guide tube into either the preoptic area (POA), retrochiasmatic area (RCh), or the median eminence region (ME). Blood samples were taken every 15 min for 2 h before and 4 h during implantation of these drugs and were analyzed for LH and prolactin by RIA. Control (no treatment) samples were obtained similarly from the same animals on another day. Placement of PBZ into the POA significantly increased LH pulse frequency and mean LH concentrations over control values whereas PIM did not. In contrast, PIM significantly increased LH pulse frequency and mean LH concentrations when placed in the ME or in the RCh, but PBZ did not. No effects of PIM on prolactin concentrations were detected. These results suggest that an NE neural system operates in the POA and that a DA system acts in the medial basal hypothalamus (RCh or ME) to suppress GnRH pulse frequency in the ovary-intact anestrous ewe.

This study examined the role of two dopaminergic (DA) cell groups, the A-14 and A-15 DA groups, in the seasonal shift in the response of LH to estradiol negative feedback in ewes. Radiofrequency lesions were placed bilaterally, in the area of the A-15 or the ventromedial A-14 cell groups of ovariectomized ewes, while control animals underwent sham neurosurgery. The effect of estrogen was tested in anestrus by analyzing LH pulse patterns before and 3 and 10 days after the insertion of estradiol implants. To evaluate the effects of these lesions on DA inhibition of LH secretion, LH pulse patterns were compared before and after an iv injection of the DA antagonist pimozide on day 3 of estradiol treatment. LH pulses were also examined in these ewes during the breeding season before and 3 days after the insertion of estradiol implants. Also, the effect of the DA receptor agonist apomorphine was tested to determine any effect of lesions on DA receptors inhibitory to LH. Lesions in either the A-14 or A-15 area decreased, but did not completely abolish, estradiol inhibition of LH pulse frequency in anestrus. Both types of lesions also blocked the stimulatory effects of pimozide on LH pulse frequency in estradiol-treated ovariectomized anestrous ewes. During the breeding season, estrogen decreased LH pulse amplitude, but not frequency, in all groups. The DA receptor agonist apomorphine decreased LH pulse frequency in all groups. Furthermore, immunohistochemistry for tyrosine hydroxylase revealed catecholaminergic fibers apparently connecting the caudal A-14 and the rostral A-15 areas. These results suggest that both the A-14 and A-15 DA cell groups are involved in the inhibition of LH by estradiol in anestrous, but not breeding season, ewes. Seasonal shifts in the activity of these DA neurons may, thus, play a role in the annual reproductive cycle of the ewe.

Evidence suggests that endogenous opioid peptides (EOP) inhibit pulsatile luteinizing hormone (LH) secretion during both the luteal and follicular phases of the ovine estrous cycle. Further data from sheep and other species indicate that the hypothalamus is the primary site of action for this EOP inhibition. The purpose of the following experiments was to determine which areas of the hypothalamus are involved in the EOP inhibition of pulsatile LH secretion. Regularly cycling ewes (n = 10) were stereotaxically implanted with guide tubes into the preoptic area (POA) and medial basal hypothalamus (MBH). Implants containing the EOP antagonist WIN 44,441–3 (WIN) were placed into each of these areas. Blood samples were collected at 12-min intervals for 3 h before and during WIN administration in the luteal phase and for 4 h before and during WIN administration in the follicular phase of the estrous cycle. During the luteal phase, WIN implants in either area increased (p < 0.01) LH pulse frequency (POA 1.4 ± 0.3/3 h before vs. 3.1 ± 0.4/3 h during; MBH 1.1 ± 0.2/3 h before vs. 2.8 ± 0.5/3 h during). There was no effect on LH pulse amplitude. In contrast, during the follicular phase, WIN implants selectively increased (p < 0.01) LH pulse frequency when implanted in the POA (3.2 ± 0.4/4 h before vs. 5.2 ± 0.6/4 h during) while increasing (p < 0.05) only LH pulse amplitude when placed in the MBH (0.7 ± 0.2 ng/ml before vs. 1.4 ± 0.3 ng/ml during). These results suggest that although EOP act in both POA and MBH to inhibit LH secretion, different populations of EOP neurons may be activated during different phases of the estrous cycle.

It is now clear that estradiol acts in the brain to inhibit LH pulse amplitude in breeding season ewes, but the neural mechanisms underlying this action of estradiol have yet to be determined. In this study, we performed four experiments to examine the role of alpha-adrenergic neurons in the control of LH pulse amplitude in the ewe. In the first experiment, implantation of the alpha-adrenergic antagonist phenoxybenzamine into the preoptic area (POA) during the follicular phase of the ovine estrous cycle significantly increased LH pulse amplitude. A similar stimulatory effect of POA implants of this antagonist was observed in estradiol-treated ovariectomized (OVX+E) ewes, but phenoxybenzamine implants had no effect in the absence of estradiol. In contrast, systemic administration of phenoxybenzamine decreased LH pulse amplitude in both OVX and OVX+E ewes. Finally, POA implants of the alpha 1-adrenergic antagonist, prazosin, increased LH pulse amplitude in OVX+E ewes, whereas the alpha 2-adrenergic antagonist, yohimbine, had no effect in these animals. These results suggest that a noradrenergic (or adrenergic) system acts in the POA via alpha 1-adrenergic receptors to inhibit LH pulse amplitude in the ewe. They also raise the possibility that this system may be involved in the negative feedback action of estradiol during the breeding season.