ABSTRACT

It was recently reported that anterior pituitary tissue transplanted to an ectopic site contains measurable amounts of dopamine and noradrenaline. To examine the possibility of local catecholaminergic control of prolactin secretion from ectopic pituitaries, pituitary grafted and sham-operated female rats were submitted to several pharmacological treatments modifying catecholamine synthesis. Administration of a single dose of α-methyl-p-tyrosine (α-MPT) significantly reduced dopamine content in the graft, while noradrenaline content was not modified. Similar changes in the contents of dopamine and noradrenaline after α-MPT administration were observed in the hypothalamus and in the in-situ pituitary in both grafted and sham-operated rats. Plasma concentrations of prolactin were increased in both grafted and sham-operated rats after administration of α-MPT. A single injection of l-3,4-dihydroxyphenylalanine (l-DOPA) increased dopamine content in the ectopic pituitary gland without altering the noradrenaline content, and produced similar effects in the hypothalamus and in-situ pituitary of grafted and control rats. Plasma prolactin concentrations were decreased by l-DOPA in both pituitary grafted and control rats. Administration of dl-treo-dihydroxyphenylserine (DOPS) increased noradrenaline content in the ectopic pituitary and reduced plasma prolactin concentrations in pituitary grafted rats. In contrast, injection of DOPS to control rats increased both hypothalamic noradrenaline content and plasma prolactin concentrations. These results suggest that dopamine and noradrenaline present in the ectopic pituitary tissue have a role in mediating prolactin release from pituitary transplants.

J. Endocr. (1987) 113, 45–49

ABSTRACT

Nine-month-old female rats bearing an ectopic pituitary gland (from a litter-mate) under the right kidney capsule since day 30 of life and their sham-operated controls, were treated with a dopamine agonist (lysuride) or antagonist (metoclopramide). Plasma prolactin and LH levels were measured by double-antibody radioimmunoassays. Vaginal smears were taken before and during the treatment periods. Eight months after the operation, a significant (P < 0·01) increase in basal prolactin levels together with a significant (P < 0·05) reduction in LH values and permanent dioestrus occurred in the grafted animals when compared with controls. Lysuride treatment resulted in a marked reduction in plasma prolactin levels both in control and grafted rats over the whole 12 days of treatment, together with a partial restoration of plasma LH levels on day 1. From day 7 onwards a depression in LH values was again observed. Oestrous cycles were partially restored at the beginning of the treatment, but after 7 days dioestrus returned. Metoclopramide administration induced a significant (P< 0·001) increase in basal prolactin levels in both grafted and control rats. Basal plasma LH values were unaffected in controls when compared with vehicle-treated animals. An increase could be seen in hyperprolactinaemic rats after 7 or 12 days of treatment however. The LH response to the administration of LH releasing hormone (LHRH) was greater in the experimental and control metoclopramide-treated rats when compared with vehicle-treated rats. Vaginal smears were not altered in the control animals but there was a significant increase in the number of oestrous smears in grafted animals given the dopamine antagonist partially restoring the cycle pattern. After LHRH administration plasma prolactin levels decreased in vehicle-treated grafted and control animals, whereas only a tendency to lower values or no modification in basal levels was observed with lysuride or metoclopramide treatments. All these data suggest that increased plasma prolactin levels cannot modify LH secretion directly. This influence may be exerted, however, through increased hypothalamic and in-situ pituitary dopamine detected in hyperprolactinaemic animals.

J. Endocr. (1984) 100, 141–148

ABSTRACT

In a group of adult Soay rams housed indoors under an artificial light cycle of alternating 16-week periods of long and short days, there was a conspicuous longterm cycle in the peripheral plasma concentrations of β-endorphin and prolactin. The levels of β-endorphin were highest under short days and lowest under long days (15-fold change), and inversely related to the changes in the plasma levels of prolactin (120-fold change). The role of dopamine in the control of β-endorphin and prolactin was investigated in a series of experiments, conducted under both long and short days, in which rams were treated with dopamine receptor agonists (dopamine and bromocriptine) and antagonists (pimozide and sulpiride). Naloxone (opioid antagonist) was also administered to assess the additional involvement of endogenous opioids.

Dopamine injected i.v. (6·6 mg/kg every 10 min) did not significantly affect the mean plasma concentrations of β-endorphin and prolactin under either long or short days. Pimozide (0·08 mg/kg i.m. every 2 h) caused a large increase in the mean plasma concentrations of β-endorphin and prolactin under long days but not short days. Naloxone (1·6 mg/kg, i.v.), administered alone or in combination with dopamine or pimozide, had no effect on the mean plasma concentrations of β-endorphin and prolactin, except under short days when, combined with pimozide, it induced an increase in the plasma concentrations of the two polypeptides.

Bromocriptine (0·06 mg/kg, s.c.) caused a significant decrease in the plasma concentrations of both β-endorphin and prolactin; this effect was most marked at the times of increased secretion (under short days for β-endorphin and under long days for prolactin). Sulpiride (0·59 mg/kg, s.c.) produced the converse effect and caused an increase in the plasma concentrations of β-endorphin and prolactin with the amplitude and duration of the effect varying with the stage of the photoperiod-induced cycle.

From these results in the Soay ram, we conclude that dopamine inhibits β-endorphin and prolactin secretion by way of D₂ receptors under both long and short days. Endogenous opioids interact with dopamine, augmenting this inhibition under short days. Differences in the acute responses in the secretion of β-endorphin and prolactin, and the inverse relationship between β-endorphin and prolactin during the cycle, indicate that different regulatory systems involving dopamine influence the two pituitary polypeptides.

Journal of Endocrinology (1990) 127, 461–469

In view of the possible involvement of catecholamines in prolactin release, and hence in lactation, four biogenic compounds, L-noradrenaline bitartrate (Sigma), dopamine HC1 (Sigma), 5-hydroxytryptamine creatinine sulphate (5-HT) (May & Baker) and melatonin (Upjohn) were tested for their effect on the yield and composition of milk in the New Zealand White rabbit. Twenty-two lactating rabbits were used. The young were separated from their mothers on the 7th day of lactation and their number adjusted to between six and eight. Litters were allowed to suckle once each morning after their mothers had received 0·5 i.u. oxytocin i.v., and daily milk yield was calculated from the difference in weight of the litter before and after suckling. On the 9th day of lactation rabbits were anaesthetized with pentobarbitone sodium B.P. i.v., and under aseptic conditions a permanent stainless steel cannula was implanted into the brain so that the tip lay in the median

ABSTRACT

The changes in hypothalamic release of dopamine and thyrotrophin-releasing hormone (TRH) into the hypophysial portal vascular system during an oestrogen-stimulated surge of prolactin in ovariectomized rats were investigated.

A single injection of 5 μg oestradiol benzoate resulted in a reliable increase in the plasma levels of prolactin during the afternoon 3 days later. Anaesthesia did not block this afternoon surge of prolactin, although its magnitude was only half of that of unanaesthetized rats. Before and during this surge, hypophysial stalk blood was collected into methanol to analyse the hypothalamic release of dopamine and TRH. Immunoreactive TRH in these methanolic extracts eluted as a single peak with the same retention time as authentic TRH on reverse-phase high performance liquid chromatography. In comparison to the morning values, levels of dopamine decreased and those of TRH increased in hypophysial stalk blood by 50 and 240% respectively. These data indicate that hypothalamic dopamine and TRH may be involved in the afternoon surge of prolactin.

Daily treatment with parachlorophenylalanine, an inhibitor of serotonin synthesis, reduced the hypothalamic release of TRH by 50%, but did not prevent the afternoon surge of prolactin and TRH induced by oestradiol benzoate.

J. Endocr. (1985) 105, 107–112

ABSTRACT

Acetylcholine is known to stimulate the secretion of growth hormone and prolactin and the efflux of ⁸⁶Rb from bovine anterior pituitary cells: dopamine prevents the stimulation of ⁸⁶Rb efflux and of prolactin but not growth hormone secretion. The sensitivity of these responses to pertussis toxin has been determined.

Treatment of bovine anterior pituitary cells in primary culture with pertussis toxin (18 h, 100 ng/ml) did not modify the stimulation of prolactin secretion by acetylcholine, but prevented its inhibition by dopamine. In lactotrophs, dopamine but not acetylcholine receptors are therefore coupled to secretion through a pertussis toxin substrate. The stimulation of ⁸⁶Rb efflux by acetylcholine was also unaffected by pertussis toxin and, again, its inhibition by dopamine was prevented.

Treatment of the cells with pertussis toxin enhanced the secretion of growth hormone in response to acetylcholine. Nitrendepine (1 μmol/l) prevented the cholinergic stimulation of growth hormone but not prolactin secretion from these cells. Acetylcholine increased the cytoplasmic calcium concentration and this rise was enhanced by treatment of the cells with pertussis toxin. Nitrendepine partially inhibited the rise in calcium caused by acetylcholine, and prevented the enhancement of the rise following pertussis toxin treatment.

Cholinergic stimulation of growth hormone therefore depends on calcium entry through nitrendepine-sensitive channels, whereas stimulation of prolactin secretion does not, and in somatotrophs a pertussis toxin substrate may limit calcium entry through these channels. These different sensitivities of somatotrophs and lactotrophs to pertussis toxin and nitrendepine may reflect differences in the properties of the predominant calcium currents in the two cell types.

J. Endocr. (1988) 116, 393–401

ABSTRACT

Thyrotrophin-releasing hormone (TRH) stimulates GH secretion in domestic fowl by actions at pituitary and central nervous system sites. The possibility that this central action might be mediated by hypothalamic catecholamines or indoleamines was therefore investigated. When TRH was administered into the lateral ventricles of anaesthetized fowl the concentration of 3,4-dihydroxyphenylacetic acid (DOPAC, a metabolite of dopamine (DA)) in the medial basal hypothalamus (MBH) was increased within 20 min. The concentrations of MBH noradrenaline (NA), DA, serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA) were, however, unaffected by the intracerebroventricular (i.c.v.) administration of TRH, although the MBH concentrations of somatostatin and TRH were concomitantly reduced. A rapid increase in DA release into MBH extracellular fluid and its metabolism to DOPAC was also observed after i.c.v. or i.v. administration of TRH, in birds in which the MBH was perfused in vivo with Ringer's solution. Microdialysate concentrations of NA, 5-HT and 5-HIAA were not, however, affected by central or peripheral injections of TRH. Diminished GH responses to i.v. TRH challenge occurred in birds pretreated with reserpine (a catecholamine depletor), α-methyl-paratyrosine (a DA synthesis inhibitor) and pimozide (a DA receptor antagonist). These results therefore provide evidence for the involvement of a hypothalamic dopaminergic pathway in the induction of GH release following the central or peripheral administration of TRH. In contrast with its inhibitory actions at peripheral sites, DA would appear to have a central stimulatory role in regulating GH release in birds.

Journal of Endocrinology (1993) 138, 225–232

ABSTRACT

Pituitary glands of grassfrog (Rana pipiens), bullfrog (Rana catesbeiana), clawed toad (Xenopus laevis) and two species of terrapin (Chrysemys picta and Pseudemys scripta) were incubated in medium containing hypothalamic extract (HE), thyrotrophin releasing hormone (TRH), somatostatin, dopamine, or combinations of these treatments. Prolactin and GH concentrations in the medium were determined by densitometry after polyacrylamide-gel electrophoretic separation. Hypothalamic extract stimulated secretion of both hormones in all species tested. Thyrotrophin releasing hormone stimulated secretion of prolactin and GH, showing a biphasic pattern of response. Dopamine had little effect alone, but inhibited HE-and TRH-stimulated release of prolactin, but not GH, in both amphibia and reptiles. Somatostatin by itself had no apparent effect on release of hormones, but it inhibited HE- and TRH-stimulated release of GH from both amphibian and reptilian pituitary glands. These results indicate that factors affecting mammals and birds also interact in the regulation of secretion of prolactin and GH in lower vertebrate species.

J. Endocr. (1984) 102, 175–180

ABSTRACT

The effect of supramaximal electric field stimulation on [³H]dopamine (DA) release by rat adrenal capsuleglomerulosa preparations was studied using a microvolume perfusion system. When the tissues were preloaded with [³H]DA, a considerable amount of [³H]DA and [³H]noradrenaline (NA) were released in response to field stimuli. Reserpinization, calcium removal or tetrodotoxin blocking of Na⁺ influx all completely inhibited the stimulation-evoked release of DA/NA, indicating that the radioactivity released is of neuronal and vesicular origin. In the adrenal cortex, a substantial proportion of tyrosine hydroxylase and dopamine-β-hydroxylase immunoreactive nerve fibres and varicosities were observed around the zona glomerulosa. DA-containing nerves were not seen in the adrenal cortex; however, the same immunocytochemical procedures clearly demonstrated dopaminergic nerve cells and fibres in the substantia nigra and the striatum respectively, and cells of the adrenal medulla. Like the NA release from noradrenergic varicosities in the zona glomerulosa, the DA release from noradrenergic endings is not subject to negative feedback modulation through DA₂ receptors since apomorphine, a DA₂-receptor agonist, and sulpiride, a selective DA₂-receptor antagonist, failed to affect the release. After in-vivo i.v. administration of [³H]DA, the glomerulosa content of DA and NA and the in-vitro release of [³H]DA and [³H]NA of zona glomerulosa both increased, indicating that the local varicose axon terminals were able to accumulate DA from the circulation, convert it into NA and release it in response to neural activity. This local arrangement of noradrenergic axon terminals, able to take up DA from the circulation and release it or convert it into NA, provides the possibility of a fine tuning of local circulation and aldosterone synthesis in the zona glomerulosa.

Journal of Endocrinology (1993) 139, 213–226

ABSTRACT

The effect of GH-releasing factor(1–44) (GRF) alone, or together with somatostatin (SRIF), dopamine (DA), vasoactive intestinal peptide (VIP) or cycloheximide was studied in a total of ten human somatotrophinomas using a static cell culture system. Growth hormone-releasing factor (2·0×10⁻⁸ mol/l) significantly (P<0·05) stimulated GH release from nine out of ten tumours over 4-h incubations, and a dose-related effect (2·0×10⁻¹⁰−2·0×10⁻⁸ mol/l) was observed in five tumours thus studied.

Repeated GRF (2·0×10⁻⁸ mol/l)-mediated GH release was seen during 96% (n = 25) of experiments performed on six tumours over 4 h and up to 27 days in culture. Growth hormone-releasing factor (2·0×10⁻⁸ mol/l) also stimulated GH release from five out of seven somatotrophinomas during 60-min incubations.

Somatostatin (6·1×10⁻⁹ mol/l) completely inhibited GRF-induced GH secretion from four tumours studied over 4 h, but in each case there was significant (P<0·05) 'rebound' of GH release from cultures exposed to both GRF and SRIF during a subsequent recovery period. Dopamine suppressed basal GH release from two out of four tumours, but in each case had a greater inhibitory effect on GRF-mediated GH release. Vasoactive intestinal peptide directly stimulated GH release from two out of three tumours, and the effects were additive to maximal stimulatory doses of GRF. Cycloheximide significantly (P< 0·01) enhanced GRF-stimulated release of GH during a 60-min incubation, but inhibited both basal and GRF-stimulated release over 4 and 8 h. We conclude that a static cell culture system can be used successfully to study the action of GRF on human somatotrophinoma tissue for up to 27 days in culture. Human somatotrophinomas seem to be only rarely unresponsive to the stimulatory action of GRF, which is itself significantly modulated by SRIF, DA and VIP. De-novo protein synthesis appears to be necessary for the full expression of GRF activity on human somatotrophinoma tissue.

J. Endocr. (1985) 105, 269–276