LH Secretion In Ovariectomized Rats Research Articles

Involvement of Neurokinin Receptors in the Control of Pulsatile Luteinizing Hormone Secretion in Rats

It has recently been shown that neurokinin B, a tachykinin, is associated with GnRH pulse generation in sheep and goats. The aim of the present study was to clarify the role of tachykinin receptors in the control of LH secretion in rats. To this end, we evaluated the effect of CS-003, an antagonist for all three neurokinin receptors (NK1, NK2 and NK3 receptors), on pulsatile LH secretion in both sexes of rats with different routes of administration. Both oral and third ventricular administration of CS-003 suppressed LH secretion in both sexes of gonadectomized animals. Furthermore, intact male rats with oral administration of CS-003 showed decreased serum testosterone levels, which might be due to suppressed LH secretion. None of the three subtype-specific neurokinin receptor antagonists showed a significant effect on LH secretion in ovariectomized rats when each antagonist was singly administered. The present results suggest that neurokinins play a role in the control of pulsatile GnRH/LH secretion via multiple neurokinin receptors in both male and female rats.

Effect of treatment with the selective oestrogen receptor modulator LY117018-HCl on pituitary sensitivity to GnRH and subsequent ovulation

The effects of LY117018-HCl (LY; a benzothiophene similar to raloxifene) were examined on various reproductive parameters in female rats. Four-day cyclic rats were treated (10:00 h on dioestrus) with LY (0.01, 0.1, 0.5, 1, 2, 4 or 16 mg kg(-1) p.o.) and assessed for ovulation at oestrus. LY inhibited ovulation at doses as low as 0.5 mg kg(-1), and ovulation did not occur at doses of 4 and 16 mg kg(-1). LY (16 mg kg(-1)) reduced wet uterine mass and LH concentrations at the time of the expected ovulatory surge. Ovulation induced by hCG in pentobarbital-treated rats was not altered by LY treatment, indicating normal ovarian sensitivity to gonadotrophins. LY, however, completely blocked the effects of oestradiol (under either negative or positive feedback modes) on LH secretion in ovariectomized rats. GnRH secretion into hypophyseal portal blood during pro-oestrus was not affected by treatment with LY, whereas the concentrations of serum LH remained reduced. Finally, treatment with LY markedly reduced pituitary sensitivity to GnRH during pro-oestrus, as it completely blocked GnRH-induced LH secretion. These results demonstrate that LY inhibits oestradiol action in the uterus and prevents ovulation in normal cyclic rats. LY-induced inhibition of ovulation is not caused by an alteration of the ovarian response to gonadotrophins or an impairment of GnRH secretion at the hypothalamus, but by a reduction in the sensitivity of gonadotrophs to the stimulatory effects of GnRH during pro-oestrus.

Effect of treatment with the selective oestrogen receptor modulator LY117018-HCl on pituitary sensitivity to GnRH and subsequent ovulation

The effects of LY117018-HCl (LY; a benzothiophene similar to raloxifene) were examined on various reproductive parameters in female rats. Four-day cyclic rats were treated (10:00 h on dioestrus) with LY (0.01, 0.1, 0.5, 1, 2, 4 or 16 mg kg(-1) p.o.) and assessed for ovulation at oestrus. LY inhibited ovulation at doses as low as 0.5 mg kg(-1), and ovulation did not occur at doses of 4 and 16 mg kg(-1). LY (16 mg kg(-1)) reduced wet uterine mass and LH concentrations at the time of the expected ovulatory surge. Ovulation induced by hCG in pentobarbital-treated rats was not altered by LY treatment, indicating normal ovarian sensitivity to gonadotrophins. LY, however, completely blocked the effects of oestradiol (under either negative or positive feedback modes) on LH secretion in ovariectomized rats. GnRH secretion into hypophyseal portal blood during pro-oestrus was not affected by treatment with LY, whereas the concentrations of serum LH remained reduced. Finally, treatment with LY markedly reduced pituitary sensitivity to GnRH during pro-oestrus, as it completely blocked GnRH-induced LH secretion. These results demonstrate that LY inhibits oestradiol action in the uterus and prevents ovulation in normal cyclic rats. LY-induced inhibition of ovulation is not caused by an alteration of the ovarian response to gonadotrophins or an impairment of GnRH secretion at the hypothalamus, but by a reduction in the sensitivity of gonadotrophs to the stimulatory effects of GnRH during pro-oestrus.

Action of AT1 subtype angiotensin II receptors of the medial preoptic area on gonadotropins and prolactin release.

This study determined the effect of the selective angiotensin II (A II) AT1 receptor subtype antagonist losartan in the medial preoptic area (MPOA) of ovariectomized rats, treated with estrogen or untreated, on the release of gonadotropins (LH and FSH) and prolactin (PRL). The MPOA is sensitive to the action of A II and contains cell bodies of neurons producing luteinizing hormone-releasing hormone and a large density of estradiol receptors. Plasma FSH was not altered in any situation. However, losartan blocked and estradiol facilitated the stimulating and inhibitory effects of A II microinjection into the MPOA on LH and PRL secretion, respectively. The results indicate that these effects are mediated by AT1 receptors in the MPOA and that estradiol may modulate them. On the other hand, losartan itself reduced LH secretion in ovariectomized rats, indicating that the increase in the secretion of this hormone, after removal of the negative feedback caused by estradiol, is due, at least in part, to the action of A II on AT1 receptors of the MPOA.

Paraventricular norepinephrine release mediates glucoprivic suppression of pulsatile luteinizing hormone secretion.

Restriction of glucose availability by 2-deoxyglucose (2DG) suppresses pulsatile LH release. The aim of the present study was to determine whether norepinephrine (NE) release in the paraventricular nucleus (PVN) is involved in the glucoprivic suppression of LH secretion in ovariectomized rats. Twelve days after ovariectomy, animals were stereotaxically implanted with a guide cannula for microdialysis in the PVN. Two days later, the PVN was perfused continuously with Ringer's solution or Ringer's solution containing a catecholamine synthesis inhibitor, alpha-methyl-p-tyrosine (100 microM), through a microdialysis probe inserted in the guide cannula 2 h before the beginning of sampling, which lasted 3 h. Blood samples were collected every 6 min through an atrial cannula, and dialysates were collected every 20 min. One hour after the beginning of sampling, 2DG (400 mg/kg BW) was administered iv through the atrial cannula. Paraventricular NE levels significantly increased immediately after 2DG injection (P < 0.05), and both mean LH concentrations and the frequency of LH pulses decreased. By contrast, when alpha-methyl-p-tyrosine was administered into the PVN, 2DG did not produce an increase in paraventricular NE, and no depression of LH secretion occurred. These results suggest that the PVN mediates the glucoprivic suppression of LH pulses via the release of NE.

Involvement of excitatory amino acidergic inputs into the median eminence in regulating pulsatile LH secretion in ovariectomized rats

Involvement of excitatory amino acidergic inputs into the median eminence in regulating pulsatile LH secretion in ovariectomized rats

Effects of passive immunization against vasoactive intestinal peptide on serum prolactin and LH levels.

Recent findings suggest that vasoactive intestinal peptide (VIP) may be a physiological regulator of prolactin secretion and may also be involved in the control of LH secretion. In the present work we have studied the effect of the blockade of endogenous VIP by means of the injection of a specific rabbit anti-VIP serum, in male and female rats with hyperprolactinemia. The administration of 0.5 ml of the VIP antiserum in ovariectomized rats given an acute or chronic treatment with 17 beta-estradiol induced a significant decrease in serum prolactin and LH levels as compared with estrogenized-control rats injected with normal rabbit serum. Anti-VIP serum also reduced serum LH levels in ovariectomized rats not treated with estrogens. The administration of the same antiserum decreased serum prolactin levels in male rats implanted with 2 anterior pituitary glands under the kidney capsule. On the other hand, the injection of the anti-VIP serum in the morning in proestrus rats brought about an increase in serum prolactin and LH levels in the afternoon of the same day. These results confirm previous data showing that VIP is a stimulator of prolactin release, and may also participate in the control of LH secretion in ovariectomized rats acting as a facilitatory factor. During proestrus however, VIP may act in an opposite way, inhibiting, rather than stimulating, prolactin and LH release.

Hyperprolactinemia Decreases Naloxone Binding in the Arcuate Nucleus of Ovariectomized Rats

Hyperprolactinemia suppresses endogenous prolactin (PRL) secretion and inhibits LH release in ovariectomized rats. Opiate peptides appear to mediate the suppressive effects of hyperprolactinemia on both endogenous PRL and LH secretion. A mechanism by which hyperprolactinemia may change the ability of opiates to influence PRL and LH is by altering the density of opiate receptors. We, therefore, examined the effect of hyperprolactinemia on the density of naloxone binding sites in hypothalamic regions that are important in the regulation of PRL and LH secretion. Female rats, ovariectomized for 4 days, were treated with ovine prolactin (oPRL) every 8 h for 2 days, and naloxone binding sites were measured using autoradiographic procedures. oPRL treatment suppressed the concentration of naloxone binding sites throughout the arcuate nucleus but had no effect in the median eminence, suprachiasmatic nucleus, and medial preoptic nucleus. There is evidence that the tuberoinfundibular dopaminergic neurons of the arcuate nucleus are directly influenced through opiate receptors. We propose that the observed decrease in the density of opiate receptors may occur on dopaminergic neurons. This theory provides an explanation for a mechanism for the suppression of endogenous PRL and LH by hyperprolactinemia: a decrease in opiate receptors will decrease opiate suppression of dopamine neurons allowing dopamine activity to increase. Increase in dopamine release are known to decrease PRL and LH secretion in ovariectomized rats. Alternatively, decreased naloxone binding may result from homologous down-regulation of receptors due to increased opiate activity. If opiate activity increases, it may directly inhibit LHRH neurons and may suppress the activity of inhibitory neurons leading to increased dopamine activity.

Evidence for reorganization of the neuroendocrine centres regulating pulsatile LH secretion in rats receiving neonatal monosodium-L-glutamate treatment.

Previous research has shown that the combination of anterior hypothalamic deafferentation (AHD) and electrolytic lesions of the anterior part of the arcuate nuclei blocks pulsatile LH secretion in ovariectomized rats, but that neither lesion alone was effective. Furthermore, the combination of AHD with arcuate lesions produced by neonatal treatment with monosodium-L-glutamate (MSG) does not block pulsatile LH release. To distinguish between the various possible reasons for this result, AHD and electrolytic lesions of the arcuate nuclei were combined in rats which had been treated neonatally with MSG or saline of equal osmolarity. One week after brain surgery, venous catheters were installed and blood samples taken at 5-min intervals for up to 3 h for assay of plasma LH. Rats treated with MSG and bearing AHD and electrolytic lesions of the arcuate nuclei continued to show pulsatile secretion of LH. Where the electrolytic lesions were posterior and dorsal to the anterior arcuate nuclei, however, LH release was blocked whether AHD was performed or not, but only in rats treated with MSG. None of the lesion combinations blocked pulsatile LH secretion in rats treated with saline. These results suggest that neonatal MSG treatment leads to a reorganization of the hypothalamus such that the role normally played by the arcuate nuclei in the regulation of pulsatile LH secretion is taken over by other hypothalamic structures.

Stimulatory or inhibitory effects of angiotensin II upon LH secretion in ovariectomized rats: a function of gonadal steroids.

The effects of intraventricular infusions of artificial cerebrospinal fluid (aCSF) or angiotensin II (AII) on LH secretion were investigated in rats that had been ovariectomized for 8 days. In untreated ovariectomized rats, the mean whole blood concentration of LH as well as the amplitude, frequency, and nadir of the LH pulses were not affected by infusion of aCSF or 15 ng AII/h, but were suppressed in a dose-dependent fashion by infusion of AII at doses of 150 or 600 ng/h. The AII receptor antagonist, saralasin, blocked the inhibitory effect of AII, demonstrating the specificity of the response to AII. In ovariectomized rats pretreated with estradiol, infusion of AII did not modify mean blood LH levels. However, in ovariectomized rats pretreated with both estradiol and progesterone, infusions of AII at 150 or 600 ng/h produced dose-dependent increases in mean LH concentrations. The results demonstrate both inhibitory and stimulatory effects of AII upon LH secretion, the direction of the effect being determined by gonadal steroids.

Secretion of luteinizing hormone in ovariectomized adult rats treated neonatally with monosodium glutamate.

We have studied the possible effects of monosodium glutamate (MSG) on LH secretion in ovariectomized rats. In experiment 1 MSG-treated and control rats were given oestradiol benzoate at noon and 72 h later half the rats in each group were given a second injection of oestradiol benzoate or progesterone. Blood samples were taken immediately before and 6 h after these i.m. injections. At 78 h there were no significant differences in plasma LH concentration measured in the two groups of rats given progesterone or in the two groups given a second injection of oestradiol benzoate although for both MSG-treated and control rats progesterone produced a significantly (P less than 0.01) greater LH surge than did oestradiol benzoate. In experiment 2 100 ul blood samples were collected at 5-min intervals for up to 3 h from MSG-treated and control rats. For rats showing more than one pulsatile discharge of LH, peak and trough values for plasma LH concentrations were no significantly influenced by MSG treatment. However the mean pulse height was significantly (P less than 0.001) greater in the MSG-treated group than in control rats. Pulsatile release stopped more quickly in the MSG-treated rats and their mean plasma LH concentration after 120 min of blood sampling was significantly (P less than 0.05) lower than that obtained in the control animals. Thus, although some aspects of LH secretion seem to be significantly different in MSG-treated rats, these effects may result from the greater sensitivity of the MSG-treated animals to experimental manipulation.

Role of alpha-adrenergic mechanism in regulating tonic luteinizing hormone release in conscious ovariectomized rats.

The role of brain catecholamines in controlling tonic LH secretion was studied in unanesthesized, freely moving, ovariectomized rats. Pretreatment with reserpine (10 mg/kg), given 24 h before the experiment, significantly suppressed pulsatile LH secretion in these rats. Intraperitoneal administration of clonidine (1 mg/kg), an alpha-adrenergic stimulating agent, significantly raised plasma LH levels in reserpine-treated rats. On the other hand, the administration of apomorphine (1.5 mg/kg), a dopamine receptor-stimulating agent, had no effect on the plasma LH levels. These findings suggest an excitatory role of noradrenaline, operating through alpha-adrenergic receptors, in pulsatile LH secretion in ovariectomized rats.

Effect of manipulation of brain dopaminergic or serotoninergic systems on basal pulsatile LH release and perisuprachiasmatic-induced suppression of pulsatile LH release in ovariectomized rats.

This study examined the possible involvement of dopamine (DA) and/or serotonin (5-HT) in regulating basal pulsatile LH release, and in mediating perisuprachiasmatic (peri-SCN)-induced suppression of pulsatile LH secretion in ovariectomized rats. Unanesthetized animals were bled continuously through jugular vein cannulae for a 1½-h prestimulation period, 1½ h of stimulation, and up to 3/4 h after stimulation. Control rats were untreated or given <i>d</i>-butaclamol, the inactive stereoisomer of <i>d</i>-butaclamol. Other groups were pretreated either with pimozide or <i>d-</i>butaclamol to block DA receptors, parachlorophenylalanine (PCPA) or metergoline to inhibit 5-HT synthesis or block 5-HT receptors, respectively, or PCPA + pimozide to affect both 5-HT and DA systems. Prior to stimulation, regardless of whether DA and/or 5-HT systems were interfered with, no significant differences existed in mean blood LH levels, LH interpulse intervals, or the magnitude or rate of increase in blood LH levels for individual LH episodes. This suggests that DA and/or 5-HT may not be essential parts of the neurotransmitter systems regulating basal pulsatile LH secretion. During peri-SCN stimulation significant decreases in mean blood LH levels occurred in all groups. This was due entirely to a lengthening in the interval between LH pulses, since no group showed a decrease in the magnitude or rate of increase in blood LH levels for those LH pulses that did occur during stimulation. These results indicate that peri-SCN-induced suppression of pulsatile LH release is not mediated by known DA and/or 5-HT neuronal inputs to this region, and suggest that the peri-SCN area may be one brain region that influences, in a suppressive manner, only the periodicity of the pulsatile LH release process in the rat.

Modulatory effects of the amygdala on oestrogen-induced LH secretion in ovariectomized rats.

An increase in LH secretion was induced in ovariectomized oestradiol benzoate-primed rats 5 h after a second injection of oestradiol benzoate. Lesions stereotaxically placed in the cortical and basomedial amygdala of steroid-primed rats abolished this rise. The results provide evidence for a facilitatory action of the amygdala upon LH release and an involvement of this region of the limbic system in oestrogen-feedback mechanisms.