Release of LH occurred in ovariectomized, oestrogen-primed rats when the medial preoptic area (mPOA) was electrically stimulated with monophasic square pulses of 1 ms duration (50 Hz, 150 μA, 15 s on and 15 s off for 30 min). Electrochemical stimulation of the anterior cingulate area applied immediately after the first 15 min period of stimulation in the mPOA completely prevented the rise in LH normally observed during the following 15 min. This effect was suppressed either by selective blockade of noradrenaline synthesis with diethyldithiocarbamate, or following systemic or intraventricular injection of the βadrenergic blocker, propranolol, whereas it did not change after systemic atropine, pimozide or phenoxybenzamine. Isoprenaline, a β-adrenergic agonist, injected into the third ventricle of rats stimulated in the mPOA mimicked the effect of the cortical stimulation, this effect was also blocked by propranolol. Intraventricular administration of propranolol or of isoprenaline had no effect on the release of LH induced by the injection of gonadotrophin releasing hormone, showing that their action is not directly on the pituitary gland. Intraventricular injection of noradrenaline, which failed to affect the release of LH induced by stimulation in the mPOA, inhibited this release when animals were pretreated with phenoxybenzamine. On the other hand, the LH-releasing potency of noradrenaline was greatly increased if the β-receptors were blocked.

From these results it may be concluded (1) that inhibition of the secretion of LH evoked by electrochemical stimulation of the anterior cingulate cortex is mediated by an adrenergic mechanism involving a β-receptor and (2) that noradrenaline exerts an inhibitory effect on the secretion of LH through a β-receptor in addition to the known facilitatory action through an α-receptor.

SUMMARY

Oestradiol benzoate (OB) injected into spayed rats induced high levels of serum prolactin (measured by radioimmunoassay) in the afternoon of the third day after the injection. When progesterone was injected into spayed rats primed 3 days before with OB, a release of prolactin was observed in the morning. These rises in serum prolactin were prevented either by treatment with p-chlorophenylalanine (PCPA), a compound which depletes 5-hydroxytryptamine (5-HT) from the brain or by injecting the 5-HT antagonist methysergide, into the third ventricle. The blockade of prolactin release by PCPA treatment of OB-injected rats was partially reversed by the administration of 5-hydroxytryptophan. These results indicate that 5-HT containing neurones mediate the effect of ovarian steroids on prolactin release. Further support of this view was provided by experiments showing that the injection of 5-HT into the third ventricle raised serum prolactin levels.

The rise in serum prolactin observed after the ovarian steroid treatment was blocked to a great extent by the injection of picrotoxin or strychnine. Picrotoxin also blocked the rise in serum prolactin induced by the injection of 5-HT into the third ventricle but failed to affect the rise in serum prolactin after i.v. injection of thyrotrophin-releasing hormone or α-methyl-p-tyrosine treatment. The results suggest that an inhibitory neuronal mechanism mediated by 5-HT neurones is involved in the release of prolactin induced by ovarian steroids.

SUMMARY

The release and synthesis of LH and FSH were studied in adenohypophyses from ovariectomized rats incubated for a period of 4 h in flasks containing 1 ml Eagle's medium. One hemipituitary was used as the experimental gland and the other half served as a control. Glands from ovariectomized untreated animals showed a spontaneous release of LH and FSH and the amount of hormones released (per mg gland) by both the hemipituitaries was not significantly different. Also the content of the hormones at the end of the incubation period was similar in both halves.

Gonadotrophin-releasing hormone (Gn-RH) added to the incubation medium stimulated the release of LH and FSH. A dose–response relationship was obtained between doses of 0·51 and 8·00 ng/ml medium. Although lower doses were required to increase the release of LH, the amount of FSH released was higher when expressed as a percentage of gland content. Pituitary glands from ovariectomized rats treated with 5 μg oestradiol benzoate 24 h before being killed showed an increase in sensitivity to Gn-RH, but the response decreased when oestrogen was injected 2 h before death. Also the addition of oestradiol-17β to the incubation medium inhibited LH and FSH release induced by Gn-RH.

Gonadotrophin-releasing hormone increased the spontaneous synthesis of LH and FSH observed in the incubated pituitaries. This effect of Gn-RH was stimulated by the injection of oestrogen into the donor animals whereas administration of oestrogen into the medium enhanced the synthesis of LH and partially inhibited that of FSH.

These results provide evidence for a dual effect of oestrogen on the release of LH and FSH induced by Gn-RH. They also show that synthesis of gonadotrophic hormones was favoured by oestrogen or by increased gonadotrophin release.

SUMMARY

The effects of interruption of amygdaloid and hippocampal afferents to the hypothalamus on ovulation and on the release of luteinizing hormone (LH) were studied in rats. Animals acutely deprived on the morning of pro-oestrus of amygdaloid influences either by transection of the stria terminalis (ST) or lesions of the amygdala, failed to show the expected ovulation. Control lesions had no effect. In contrast, rats with long-term transection of either the ST, the cortico-hypothalamic tract (CHT), or both, had regular oestrus cycles and ovulated normally. Studies of ovulation which was induced after the administration of ovarian steroids revealed a differential response to oestrogen and progesterone. In animals with long-term transection of the ST the ovulatory response to oestrogen was similar to that induced in non-lesioned rats but fewer ovulations were induced by progesterone administration. The decreased ovulatory response after progesterone treatment was attributed to activation of inhibitory influences originating in the hippocampus, since administration of picrotoxin or section of the fornix re-established a normal response. Long-term interruption of the limbic efferents did not alter the increased levels of plasma LH found after ovariectomy. However, the release of LH induced by progesterone in ovariectomized rats primed with gonadal steroids was decreased after transection of the ST and increased after interruption of the CHT. These results are consistent with the concept that the amygdala and the hippocampus exert a modulating influence on gonadotrophin release; the influence of the amygdala being facilitatory and that of the hippocampus inhibitory.

SUMMARY

The melanocyte-stimulating hormone (MSH) content of whole toad pituitary glands decreased upon treatment with reserpine. With daily injections the values remained low for 1 week but regained normal levels after 2 weeks in spite of an increased secretion of MSH as indicated by darkening of the skin. In rats a drop in pituitary MSH content also occurred after reserpine injection but normal values were found after 7 and 14 days of treatment.

MSH-releasing factor found in stalk-median eminence tissue of normal male rats was not present in the reserpine-injected animals, but after 7 days of treatment an increase in MSH-release-inhibiting factor (MSH-R-IF) was demonstrated. MSH-R-IF was also found to have increased in female castrated rats after 2 days of treatment with reserpine.

It is concluded that reserpine permits the secretion of pituitary MSH by blocking the release of MSH-R-IF, which accumulates in the hypothalamic neurones.

SUMMARY

Different regions of the midbrain were stimulated by electrolytic deposition of iron from stainless steel unipolar electrodes. Electrochemical stimulation of the ventral tegmental area (VTA), the raphe nuclei or the peri-aqueductal grey in rats in pro-oestrus was effective in preventing spontaneous ovulation. No blockade of ovulation was observed by stimulating other mesencephalic structures. After stimulation in the VTA there was a significant decrease of the elevated serum levels of luteinizing hormone (LH) found after an injection of progesterone into pro-oestrous rats or into ovariectomized—oestrogen-primed animals. In contrast, electrochemical stimulation of the dorsal mesencephalic tegmentum, lateral and inferior to the peri-aqueductal grey, in rats in which spontaneous ovulation had been blocked by continuous illumination, resulted in an ovulatory response. Serum LH was found to increase in gonadectomized—oestrogen-treated male and female rats after stimulating this area. No significant changes in serum follicle-stimulating hormone were found after stimulation in the ventral or dorsal tegmentum. It is concluded that both stimulatory and inhibitory influences on the release of gonadotrophin can be evoked by stimulating mesencephalic structures. The stimulatory and inhibitory effects could depend on the activation of noradrenergic and serotoninergic systems, respectively.

SUMMARY

Serum follicle-stimulating hormone (FSH) concentration was found to be higher in neonatal female rats than in adults. The values increased to a maximum on day 15 and decreased thereafter. Ovariectomy soon after birth or at 5 days of age induced a significant rise in serum FSH concentration 9 days later. Administration of 10 μg oestradiol benzoate (OB) lowered FSH concentration in both intact and spayed animals.

Progesterone (1 mg) injected 3 days after priming female rats with a single dose of 10 μg OB induced, on the same day, a significant rise in serum FSH concentration in animals older than 22 days of age. In younger animals progesterone reversed the effect of OB. The facilitatory effect of progesterone occurred when the hormone was given in the afternoon but not when it was given in the morning.

Male rats, although showing the negative feedback effect of OB injection, failed to show the positive feedback effect of progesterone.

It is concluded that the central nervous system—hypophysial mechanism responsible for FSH secretion is ready to function before puberty. In female rats initiation of puberty probably depends on the activation of that mechanism by appropriate facilitatory ovarian steroid signals.

SUMMARY

In immature female rats serum luteinizing hormone (LH) concentration, as measured by radioimmunoassay, was found to be higher at 10 or 15 days of age than thereafter. Animals ovariectomized soon after birth or at 5 days of age showed a significant rise in serum LH levels 10 days later.

A positive feedback effect on LH secretion was observed on the day following a single injection of oestradiol benzoate (OB) in 28-day-old rats but not in younger animals. However, in animals primed with OB a second dose of OB 2 days later resulted in a significant rise in serum LH levels even in rats of 22 days of age.

Progesterone (1 mg) injected 3 days after the injection of a single dose of OB induced, a few hours later, a significant rise in serum LH concentration. This effect was observed from the 22nd day of age but not in younger animals. The magnitude of the response to progesterone, as revealed by the serum LH levels, sharply decreased at the time of puberty.

It is concluded that the mechanisms responsible for the tonic release of LH are ready to function at the time of birth or shortly thereafter, while those involved in the phasic release mature around 22 days of age.

The effect of frontal hypothalamic deafferentation on the release of LH and FSH was studied in ovariectomized rats. Frontal cuts were placed just in front of the arcuate nucleus, at the posterior border of the optic chiasma (RCS), at the level of the anterior commissure (POS) and in front of the optic chiasma (PCS). Animals with RCS and POS cuts showed vaginal smears with persistent cornification; the other groups had irregular cycles.

The concentrations of LH and FSH in the serum increased after ovariectomy in deafferentated animals, but after 4 weeks the levels were lower than in the animals without hypothalamic lesions except for the PCS group. The more caudally that the cuts were located, the lower were the concentrations of hormones in the serum. The injection of repeated doses of oestradiol benzoate resulted in a decrease in serum gonadotrophin of both rats without hypothalamic lesions and RCS rats. Although a greater decrease was observed in the lesioned than in the intact rats, it is believed that such an effect does not indicate an increased sensitivity of deafferentated animals to this steroid.

The stimulatory effect of progesterone on LH and FSH release was studied in ovariectomized rats primed with oestradiol benzoate. The responses were unchanged in PCS animals but failed to occur in POS and RCS rats. Measurement of the level of gonadotrophin-releasing hormone in frontal hypothalamic slices from RCS animals showed a decreased level behind the cut and an increased one in front of it, suggesting that perikarya located in front of the section were sending their axons to the mediobasal hypothalamus. It is believed that the blockade of the stimulatory effect on gonadotrophins by frontal hypothalamic deafferentation is due to the transection of these axons. Cuts placed immediately in front of the arcuate nucleus, however, permitted progesterone-induced gonadotrophin release because of incoming neurones containing gonadotrophin-releasing hormone, which end in structures immediately rostral to the cut. The results indicate that effects of both inhibitory and stimulatory ovarian steroid feedback are impaired by frontal hypothalamic deafferentation.

SUMMARY

The concentration of prolactin in serum after oestrogen and progesterone injection into spayed rats was measured by radioimmunoassay.

After a single injection of 5 μg oestradiol benzoate (OB) into long-term ovariectomized rats, serum prolactin concentrations showed a circadian rhythm with high levels in the afternoon and almost no changes in the morning. Peaks of prolactin occurred 2, 3 and 4 days after the injection. Below a dose of 1 μg OB, the response was dose-dependent, but the response was then maximal.

In spayed rats primed with 5 μg OB, the injection of 2 mg progesterone 2, 3 or 4 days later resulted in a significant increase in serum prolactin. This response, in contrast to that of oestrogen, occurred in the morning and in the evening and was found to be dose-dependent. The rise in serum prolactin after injection of 1 mg progesterone also showed a close relationship to the priming dose of OB. Progesterone had no effect in spayed, untreated animals. Maximal levels of prolactin were attained 3–4 h after the s.c. injection of progesterone. The release of prolactin which can be induced either by OB or by progesterone was blocked by the administration of progesterone injected 1 day before the expected release would occur. These results indicate that progesterone exerts both facilitatory and inhibitory effects on prolactin secretion. Male rats were found to be less sensitive to the ovarian steroid treatment.

It is suggested that oestrogen could be responsible for the rise in prolactin observed at pro-oestrus and progesterone for the increase in prolactin in pseudopregnancy and pregnancy.