ABSTRACT

The concentrations of dopamine, noradrenaline and their respective primary neuronal metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and 3,4-dihydroxyphenylethyleneglycol (DHPG) were measured in the hypophysial portal and peripheral plasma of sheep and rats by combined gas chromatography–mass spectrometry. Hypophysial portal and jugular blood samples were taken at 5- to 10-min intervals for 3–7 h from six conscious ovariectomized ewes. Blood was also collected for 30 min under urethane anaesthesia from the cut pituitary stalk from 16 pro-oestrous female and five intact male rats.

In ovariectomized ewes, noradrenaline concentrations were higher in hypophysial portal plasma than in peripheral plasma (6·6 ± 0·8 vs 2·2 ± 0·4 nmol/l). In contrast, dopamine was undetectable (<1 nmol/l) in the portal and peripheral plasma of all ewes. Plasma levels of DOPAC and DHPG in portal and jugular samples were similar. In all pro-oestrous female rats, plasma concentrations of dopamine were higher in portal blood than in jugular blood (8·0±1·4 vs 4·8± 0·6 nmol/l). Detectable concentrations of dopamine were measured in the portal plasma of two out of five male rats. Noradrenaline concentrations were higher in portal plasma than in peripheral plasma of both female (8·3 ± 1·7 vs 3·7 ± 0·6 nmol/l) and male (14·8± 2·7 vs 6·1± 1·2 nmol/l) rats.

These data show that noradrenaline, but not dopamine, is secreted into the long portal vessels in sheep. The results suggest that there are species differences in the secretion of hypothalamic dopamine into hypophysial portal blood.

Journal of Endocrinology (1989) 121, 141–147

The changes in adenohypophysial and hypothalamic content and in hypothalamic release of dopamine and thyrotrophin-releasing hormone (TRH) into the hypophysial portal system during the suckling-induced release of prolactin were investigated. An increase in peripheral plasma levels of prolactin was induced by mammary nerve stimulation in urethane-anaesthetized and by suckling in unanaesthetized lactating rats. In the unanaesthetized rat, suckling caused a decrease of dopamine levels in hypothalamus and adenohypophysis and a short-lasting small increase in hypothalamic TRH. Mammary nerve stimulation induced a transient decrease in dopamine levels and an increase in TRH levels in hypophysial stalk blood. To assess the significance of the observed changes in dopamine and TRH levels for prolactin release, these changes in dopamine and TRH were mimicked in lactating rats anaesthetized with urethane and pretreated with α-methyl-p-tyrosine (AMpT, a competitive inhibitor of catecholamine synthesis). Reducing hypothalamic dopamine secretion by treatment with AMpT increased peripheral plasma levels of prolactin from 15 to 477 ng/ml; an infusion with dopamine, resulting in plasma levels similar to those measured in hypophysial stalk plasma, reduced plasma levels of prolactin to 127 ng/ml. Neither a 50% reduction in dopamine infusion rate for 15 min nor administration of 100 ng TRH caused an appreciable change in plasma prolactin levels. However, when dopamine infusion was reduced by 50% for 15 min just before TRH was injected, then an increase in plasma levels of prolactin from 172 to 492 ng/ml was observed. Thus, the effectiveness of TRH in releasing prolactin in the lactating rat was enhanced when a transient decrease of dopamine levels occurred before treatment with TRH. It is concluded that the changes observed in dopamine and TRH levels in hypophysial stalk blood are involved in the suckling-induced prolactin release in an important manner.

ABSTRACT

The effects of dopamine on GH release were investigated both in vivo in freely moving intact rats and in rats with a mediobasal hypothalamic lesion, and in vitro in a perifusion system using dispersed male rat pituitary cells kept in primary culture. In vivo, dopamine (5 mg/kg body weight) induced a rapid and very transient increase in plasma GH levels in lesioned but not in intact rats. This increase was markedly inhibited by a prior injection of the D₁ antagonist SCH 23390 (0·5 mg/kg) but not of the D₂ antagonist domperidone (0·5 mg/kg). The D, agonist SKF 38393 induced a dose-dependent stimulation of GH release in lesioned rats, and the effect obtained with a dose of 5 mg/kg was abolished by pretreatment with SCH 23390 (0·5 mg/kg). In vitro, dopamine (0·1 μmol/l) and SKF 38393 (0·1 μmol/l) provoked a rapid and reversible release of GH from superfused rat pituitary cells; this effect was markedly inhibited by simultaneous superfusion of SCH 23390 (1 μmol/l). These findings indicate that dopamine can stimulate basal GH release at the pituitary level and that this stimulation is mediated by D₁ but not by D₂ receptors. They also support the hypothesis that unidentified hypothalamic neurohormones may modulate this effect.

Journal of Endocrinology (1990) 127, 191–196

The effects of dopamine and thyrotrophin-releasing hormone (TRH) on prolactin release was studied in 14 intact and six pituitary stalk-sectioned (SS) female rhesus monkeys (Macaca mulatta). Baseline prolactin values were ninefold higher in SS animals (149 ± 16 ng/ml) than in intact animals (16 ± 1 ng/ml).

Prolactin release after intravenous administration of TRH in doses of 0,125,250, 500 and 1000 ng revealed that SS monkeys were more sensitive to the prolactin-releasing activity of this tripeptide than were intact animals. A significant (P < 0·05) increment in serum prolactin was observed in SS animals after injection of 125 ng TRH whereas 250 ng was required to raise prolactin levels in the circulation of intact animals significantly (P <0·05). Furthermore, at each comparable dose level of TRH, the increment in serum prolactin was distinctly greater in SS animals than in intact monkeys.

Infusion of dopamine at the rate of 10 μg/kg body weight per min significantly (P <0·05) lowered prolactin levels within 60 min in intact animals and no further decline was observed with 20 or 40 μg dopamine. Serum prolactin concentrations were not affected by saline infusion or by 5 μg dopamine. Infusion of dopamine at the rate of 10 μg/kg body wt per min also resulted in significant (P <0·01) suppression of serum prolactin in SS animals. This prolactin decrease was apparent within 40 min. Prolactin release after 500 ng TRH was less in these dopamine-treated SS monkeys than after an infusion of saline. Higher doses of dopamine (20 and 40 μg) did not cause a further decrease in basal serum prolactin concentrations, but these two dopamine treatments blocked the increase in prolactin elicited by 500 ng TRH.

The results suggest that the removal of hypothalamic influence, possibly related to the effects of dopamine, renders the pituitary gland more sensitive to the prolactin-releasing action of TRH.

Department of Experimental Endocrinology, Institute of Endocrinology, Medical Academy of Łódź, Dr Sterling sir. 3, 91–425 Łódź, Poland

(Received 22 May 1978)

Lloyd, Meares & Jacobi (1975) observed inhibition of mitotic activity in the anterior pituitary gland by the dopamine receptor agonist, bromocriptine, in oestrogen-treated male rats. This observation has been confirmed in our laboratory (Stępień, Wolaniuk & Pawlikowski, 1978). Suppression of mitotic activity in the pars intermedia of the rat pituitary gland by bromocriptine has also been observed (Rychter & Stępień, 1977). Furthermore, it has been found that the dopamine receptor blocker, pimozide, enhances mitotic activity in the rat anterior pituitary gland (Stępień et al. 1978). By the use of various ergot alkaloids, MacLeod & Lehmeyer (1973) succeeded in inhibiting the growth of transplantable rat pituitary tumours. There have also been observations suggesting an antiproliferative effect of bromocriptine on human pituitary tumours (Wass, Thorner, Morris, Rees, Mason, Jones &

SUMMARY

Cyclic female rats were treated with reserpine (5 mg/kg) on the 2nd day of dioestrus (15.00 h) or on the day of pro-oestrus (11.30 h) to produce a total or partial depletion of brain amines. Ovulation was inhibited in 18 out of 21 rats. Intrahypothalamic infusions of 0·4–80 μg dopamine or noradrenaline on the afternoon of pro-oestrus did not restore ovulation in reserpine-treated rats. In normal pro-oestrous rats, 80 μg dopamine significantly inhibited ovulation, whereas 80 μg noradrenaline had no significant effect. Electrochemical stimulation of the basal hypothalamus or the administration of luteinizing hormone induced ovulation only in those animals treated with reserpine at pro-oestrus. The results indicate that reserpine exerts a marked peripheral effect when given on the 2nd day of dioestrus and has a partial effect on the ovaries when administered at pro-oestrus. The failure of monoamines to induce ovulation suggests that some aminergic synapses involved in gonadotrophin release may lie outside the medial basal hypothalamus. In addition, high levels of dopamine appear to exert an inhibitory action on the release of pituitary gonadotrophins.

ABSTRACT

Dopamine acts directly on the pituitary to modulate gonadotrophin (GtH) secretion in goldfish (Carassius auratus). In the light of this important role for dopamine in the regulation of goldfish reproduction, this investigation was designed to evaluate the receptor specificity of this dopamine inhibition and to describe the use of domperidone, a specific dopamine D₂-receptor antagonist, in the manipulation of pituitary function in goldfish. To investigate the specificity of dopamine inhibition of GtH secretion, selected dopamine receptor antagonists were injected i.p. to block dopamine receptors thereby increasing GtH secretion as reflected by increased serum concentrations of GtH. Serum GtH levels were significantly increased by the active stereoisomer (−)-sulpiride in a dose-related fashion; (+)-sulpiride had no effect. Comparison of dopamine antagonists at low doses indicated that only domperidone and pimozide caused significant increases in serum concentrations of GtH. Dopamine antagonists potentiated the action of a gonadotrophin-releasing hormone analogue (GnRH-A) with an order of potency of domperidone = pimozide > metoclopramide = fluphenazine. [³H]Domperidone, injected i.p. with unlabelled domperidone, entered the blood and achieved maximum concentrations 12 h after injection, but did not accumulate in the brain in appreciable amounts. Gonadal ³H radioactivity was usually equal to or in excess of blood radioactivity, while [³H]domperidone was highly concentrated in the pituitary in a time-dependent fashion, with maximal accumulation occurring 24 h after injection. The time-course of pituitary accumulation of [³H]domperidone correlated well with the temporal increase in serum GtH levels in response to i.p. injected domperidone or domperidone plus an analogue of LHRH. Domperidone increased serum concentrations of GtH in a dose-related fashion; an analogue of salmon GnRH (sGnRH-A) increased the sensitivity and magnitude of the serum GtH response to domperidone. Serum concentrations of GtH were increased by sGnRH-A in a dose-related fashion; a low dose of domperidone substantially increased the sensitivity of the serum GtH response to sGnRH-A.

These results indicate that dopamine inhibits GtH secretion from the goldfish pituitary by acting through a specific mechanism mediated by a dopamine D₂ receptor. Domperidone increased serum concentrations of GtH, potentiated the action of gonadotrophin-releasing hormones and did not pass into the brain after i.p. injection into goldfish. The data also suggest that dopamine and GnRH, although acting through different receptors, influence the effect of each other on GtH release.

J. Endocr. (1987) 114, 449–458

ABSTRACT

We have studied the effect of dopamine together with agonist and antagonist drugs of different specificities on the release of TRH from the perfused, intact hypothalamus of the adult rat in vitro. Dopamine produced a dose-related stimulatory effect on TRH release with maximal effect being achieved at 1 μmol/l (increase over basal, 118 ±16·5 (s.e.m.) fmol TRH; P <0·001 vs basal). This effect was mimicked by the specific D₂-agonist drugs bromocriptine (0·1 μmol/l) and LY 171555 (0·1 μmol/l) (increase over basal values, 137·5±13·75 fmol and 158·6± 10·7 fmol respectively; P <0·001 vs basal), but not by the D₁-agonist SKF 38393A. The stimulatory effect of dopamine (1 μmol/l) was blocked in a stereospecific manner by the active (d) but not by the inactive (l) isomers of the dopamine antagonist butaclamol. Similar blockade was achieved with the specific D₂-antagonist domperidone (0·01 μmol/l) whereas the D₁-antagonist SCH 23390 was only effective when used at a concentration 100 times greater. Lower concentrations (0·01 μmol/l) of this D₁ -antagonist did not block the stimulatory effect of dopamine. High-performance liquid chromatography characterization of the material secreted within the hypothalamus showed one single peak of immunoreactive material which coeluted with synthetic TRH. These data suggest that dopamine exerts a stimulatory role in the control of hypothalamic TRH release by acting at specific D₂-receptors.

J. Endocr. (1987) 115, 419–424

ABSTRACT

Dopamine and morphine modulate GH and LH release, probably at a hypothalamic locus. To investigate this in more detail, we studied the influence of these substances on somatostatin and LH-releasing hormone (LHRH) release from rat hypothalamic fragments in vitro. Hypothalamic fragments were incubated in Earle's medium. After 60 min of preincubation, medium from two 20-min incubations was collected and somatostatin and LHRH levels measured by radioimmunoassay. Dopamine (10 nmol/l–0·1 mmol/l) induced a progressive increase (r = 0·41; P <0·01) in basal somatostatin levels. K ⁺ (30 mmol/l)-induced somatostatin release was also increased (r = 0·54; P <0·01) by increasing doses of dopamine. Metoclopramide (10 μmol/l) blocked the dopamine (1 μmol/l)-induced increase in somatostatin release. No significant relationship between dopamine and LHRH was found either basally or after K ⁺ (30 mmol/l) stimulation. Basal somatostatin was negatively correlated (r = −0·63; P <0·01) with morphine concentrations. No significant correlation was found after K⁺ (30 mmol/l) depolarization. Basal LHRH release was not influenced by morphine, while K ⁺(30 mmol/l)-induced release was significantly lower than controls only at a concentration of 10 nmol/l. These results suggest that dopamine and morphine act at a hypothalamic level to modulate GH release through alterations in somatostatin secretion. Dopamine and morphine have no consistent effect on hypothalamic LHRH release.

J. Endocr. (1985) 106, 317–322

ABSTRACT

Experiments were carried out on the antagonistic effects of opiates on the inhibition by dopamine of prolactin secretion from rat anterior pituitary glands. Dose–response and time-course experiments were carried out using both static incubation of paired hemipituitary glands and perifusion of whole glands. Dopamine (10–1000 nmol/l) was found to have an inhibitory effect on prolactin secretion, but at a lower concentration (0·1 nmol/l) a small stimulation was observed. Against an inhibition established with 100 nmol dopamine/l in static incubation, the three opiates under study, morphine sulphate, Leu⁵enkephalin and d-Ala²,Met⁵-enkephalin (DAME), had a maximum antagonistic effect at 50–1000 nmol/l in a 90-min incubation. Morphine and DAME were rather more effective than Leu⁵-enkephalin, possibly because of degradation of the latter. Naloxone reversed the effect of morphine. All three opiates showed little effect on dopamine-inhibited prolactin secretion in a perifusion system. The data accord with previous suggestions that prolactin secretion may be stimulated both by very low concentrations of dopamine and by opiates acting to reverse the inhibition exerted by higher dopamine concentrations. It should be noted that both morphine and the enkephalins have similar effects on prolactin secretion, despite their normal specificity for different opiate receptors; their actions on the pituitary may thus be rather non-specific.

J. Endocr. (1986) 109, 313–320