Estrogen inhibition of dopamine release into hypophysial portal blood

The hypothesis that 17 beta-estradiol suppresses dopamine secretion into hypophysial portal blood was tested. Portal plasma concentrations of dopamine were significantly lower in proestrous rats (1.0 +/- 0.1 ng/ml; mean +/- SE) than in estrous rats (1.9 +/- 0.38 ng/ml). To deplete the animal of endogenous steroid hormones, proestrous rats were adrenalectomized (Adx) and ovariectomized (Ovx). Twenty-four hours later, hypophysial portal blood was collected for 60 min, and the plasma from this blood was analyzed for dopamine. Arterial plasma from these rats was assayed for 17 beta-estradiol and progesterone. The concentrations of dopamine in the portal plasma of sham-operated rats and bilaterally Adx-Ovx rats were similar to those in estrous animals. The concentration of dopamine in portal plasma of Adx-Ovs rats injected 24 h earlier with 50 micrograms 17 beta-estradiol was 1.0 +/- 0.31 ng/ml, which was comparable to that in proestrous animals but less than that in the estrous rats. The concentrations of 17 beta-estradiol in arterial plasma were as follows: 24 +/- 8.3 pg/ml in proestrous rats, 40 +/- 2.9 pg/ml in estrous rats, 10 +/- 1.3 pg/ml in Adx-ovx rats, and 96 +/- 17.3 pg/ml in Adx-Ovx rats injected with 50 micrograms 17 beta-estradiol. Twenty-four hours after injection of 25 micrograms 17beta-extradiol into Adx-Ovx rats, the plasma 17beta-estradiol levels were 51 +/- 7.4 pg/ml, and the dopamine concentrations in portal plasma were 1.9 +/- 0.57 ng/ml. It is concluded that an acute effect of 17 beta-estradiol is suppression of hypothalamic secretion of dopamine into hypophysial portal blood.     

Sex-related difference in the release of dopamine into hypophysial portal blood

The concentration of dopamine (DA) in pituitary stalk plasma of cycling female rats during diestrus was approximately 7 times that in stalk plasma of intact male rats, and the rate of DA synthesis in the median eminence of diestrous female rats was 5 times that in the median eminence of intact male rats. DA concentrations in pituitary stalk plasma of castrated adult male rats, orchiectomized as adults or as 1-day-old neonates, did not differ significantly from those of intact adult male rats. However, treatment of male rats with 17 beta-estradiol benzoate for 3 days resulted in a significant (P less than 0.005) increase in the concentration of DA in pituitary stalk plasma. DA concentrations in stalk plasma of adult female rats, ovariectomized as adults or treated with testosterone propionate (50 micrograms) on day 1 of life, did not differ appreciably from those of diestrous female rats. However, DA concentrations in stalk plasma of adult female rats that had been ovariectomized on day 14 of life were significantly (P less than 0.01) lower than those of diestrous female rats. In view of these results, it is concluded 1) that there is a sex-related difference in the release of DA from tuberoinfundibular neurons into hypophysial portal blood, and 2) that this difference is not due to a suppressive action of androgen on the secretion of DA in the male rat, but is a consequence of a stimulatory action of estrogen on the release of DA in the female rat.     

Immunoreactive somatostatin in rat hypophysial portal blood.

Somatostatin levels have been determined by RIA in hypophysial portal blood of pentobarbital-anesthetized male rats. In most animals, immunoreactive somatostatin (SRIF) levels were higher in hypophysial portal blood than in systemic blood. In euthyroid rats, the mean level was 158 +/- 27 pg/ml (n = 8); SRIF was undetectable (less than 30 pg/ml) in systemic blood of these rats. It is suggested that endogenous SRIF was not degraded during the collection of stalk blood, since synthetic SRIF is stable when incubated in rat serum during 4 min at 37 c and 2 h at 0 C, i.e. under the conditions the blood was kept during the collection. SRIF in hypophysial portal plasma had the same immunoreactivity with a specific antiserum against SRIF as did synthetic SRIF. Gel filtration of hypophysial portal plasma revealed two immunoreactive peaks, the major one corresponding to synthetic SRIF, the smaller one representing a larger molecular form. Thyroidectomy and excess of T4 did not modify the levels of SRIF in hypophysial portal blood, suggestinc SRIF is stable when incubated in rat serum during 4 min at 37 C and 2 h at 0 C, i.e. under the conditions the blood was kept during the collection. SRIF in hypophysial portal plasma had the same immunoreactivity with a specific antiserum against SRIF as did synthetic SRIF. Gel filtration of hypophysial portal plasma revealed two immunoreactive peaks, the major one corresponding to synthetic SRIF, the smaller one representing a large molecular form. Thyroidectomy and excess of T4 did not modify the levels of SRIF in hypophysial portal blood, suggesting that the feedback of thyroid hormones on TSH secretion does not involve changes in the secretion of SRIF by the hypothalamus.     

Stimulation of dopamine release into hypophysial portal blood by administration of progesterone

To test the hypothesis that progesterone is involved in the regulation of dopamine release into hypophysial portal blood, female rats were adrenalectomized and ovariectomized at 1200 h on the day of proestrus and, immediately after the operation, injected sc with sesame oil or progesterone. Approximately 24 h later, hypophysial portal blood was collected and the plasma from this blood was analyzed for dopamine. The concentration of dopamine in portal plasma from rats given sesame oil or progesterone (50 mg) was 1.8 +/- 0.24 or 6.0 +/- 1.2 ng/ml (mean +/- SE), respectively. At the time of collection of portal blood, the progesterone concentration in arterial plasma was less than 0.8 ng/ml in rats given sesame oil and was 93 +/- 11 ng/ml in animals given progesterone. Conversely, the circulating concentrations of PRL was less (25 +/- 3 ng/ml) in animals given progesterone and higher (42 +/- 5 ng/ml) in animals given sesame oil. Twenty-four hours after the injection of progesterone (50 mg) into intact proestrous rats, the dopamine concentration in portal plasma was 5.8 +4- 1.29 ng/ml, a concentration which was significantly (P less thann 0.01) higher than that in untreated proestrous rats (1.1 +/- 0.24 ng/ml) but only slightly higher than that in untreated estrous rats (3.7 +/- 1.0 ng/ml). It is proposed 1) that progesterone administered sc can lead to an increased secretion of dopamine into hypophysial portal blood and 2) that the increased concentration of dopamine in portal plasma of intact pregnant rats relative to that of proestrous rats is, in part, a consequence of augmented progesterone secretion.     

Immunoreactive somatostatin in rat hypophysial portal blood (author's transl)

Somatostatin levels have been determined by radioimmunoassay in hypophysial portal blood of pentobarbital-anesthetized male rats. In euthyroid rats, the mean level was 158 +/- 27 pg/ml (n = 8); somatostatin was undetectable (less than 30 pg/ml) in systemic blood of these rats. Thyroidectomy and excess of T4 did not modify the levels of somatostatin in hypophysial portal blood.     

Epinephrine in rat hypophysial portal blood is derived mainly from the adrenal medulla

This study was performed to determine if epinephrine (EPI) present in hypophysial portal blood has a peripheral or a central origin. Pituitary stalk and femoral arterial EPI plasma levels were simultaneously measured in rats anesthetized with thiopental or urethane. EPI was measured by two different methods: radioenzymatic assay or HPLC followed by electrochemical detection. In thiopental-anesthetized rats, levels were higher in hypophysial portal than in femoral arterial plasma with both methods. In urethane-anesthetized rats, no difference in EPI concentrations was found between peripheral and portal plasma. A rapid blood volume depletion (15 ml/kg in 3 min) evoked a marked elevation of pituitary stalk and arterial EPI plasma concentration while norepinephrine (NE) and dopamine (DA) levels were unaffected. Under both types of anesthesia, the increase of EPI levels was higher in peripheral than in portal plasma. After removal of the adrenal glands, EPI was undetectable or barely detectable in pituitary stalk and arterial plasma. The hemorrhage-induced stimulation of EPI secretion was blunted in portal and peripheral plasma of adrenalectomized rats. These findings indicate that: (1) hypophysial portal EPI plasma concentration is highly dependent on an adrenal source; (2) hypothalamic EPI is not released into portal circulation in response to acute hemorrhage, and (3) anesthesia markedly influences catecholamines plasma levels.     

Corticotropin-releasing factor secretion increases in rat hypophysial portal blood during insulin-induced hypoglycemia

To study the possible involvement of hypothalamic corticotropin-releasing factor (CRF) in the stimulation of adrenocorticotropic hormone (ACTH) release caused by insulin-induced hypoglycemia (IIH), we measured CRF secretion in hypophysial portal blood (HPB) in rats anesthetized with sodium thiopental after injection of insulin. Before treatment, systemic ACTH levels (952 +/- SE 143 pg/ml; n = 12) were well above normal values, probably reflecting the anesthetic and surgical stress consecutive to the preparation for portal blood collection. Insulin injection induced a significant increase of ACTH release within 15 min (1,588 +/- 168 vs. 741 +/- 144 pg/ml; n = 6, in vehicle-injected rats) which lasted for at least 1 h. CRF levels in HPB were 857 +/- SE 168 pg/ml (n = 13) during the first-hour pretreatment collection. Vehicle injection did not modify CRF secretion (759 +/- 142 pg/ml; n = 6). Insulin injection provoked a significant increase in CRF release (1,449 +/- 257 pg/ml; n = 7). These data suggest that an increased hypothalamic CRF secretion is responsible for the stimulation of pituitary ACTH release following IIH. The possible involvement of central neuromediators in the IIH-induced CRF production is discussed.     

Stimulation by bombesin of immunoreactive somatostatin release into rat hypophysial portal blood

Bombesin was injected into the cerebral ventricle of male rats anesthetized with urethane to study its effect on plasma levels of immunoreactive somatostatin (IRS) in hypophysial portal and jugular blood. An intraventricular injection of bombesin (0.2 and 2 micrograms/rat) caused a significant and dose-related increase in plasma IRS in hypophysial portal blood but not in jugular blood. Although bombesin placed into the cerebral ventricle is known to stimulate glucagon and epinephrine release, an iv injection of glucagon (100 micrograms/100 g BW) or epinephrine (2.5 micrograms/100 g BW) did not cause any significant changes in plasma IRS levels in hypophysial portal and jugular blood, suggesting that these substances do not mediate bombesin stimulation of portal IRS release. Pretreatment with naloxone (75 micrograms/100 g BW, iv) failed to affect the portal IRS release induced by bombesin (2 micrograms/rat), indicating that the opiate receptor is not likely to be involved in this reaction. To ascertain whether IRS released by bombesin into hypophysial portal blood is biologically active, the effect of bombesin on the plasma GH level was then examined. Bombesin (2 micrograms/rat) injected intraventricularly completely suppressed the rise of plasma GH after the intraventricular injection of beta-endorphin (1 microgram/rat) or the iv injection of prostaglandin E1 (5 micrograms/100 g BW). Bombesin thus appears to stimulate the secretion of IRS, and probably biologically active somatostatin as well, from the hypothalamus into hypophysial portal blood, thereby inhibiting GH release from the anterior pituitary.     

Relationship between dopamine release into hypophysial portal blood and prolactin release after morphine treatment in rats

To address the issue of whether, after morphine treatment, the reduced release of dopamine (DA) into portal blood is entirely responsible for the increased prolactin (PRL) release, the following study was conducted. The concentration of DA in plasma from a single portal vessel of untreated, ovariectomized rats was 1.85 +/- 0.33 ng/ml (mean +/- SE). Treatment of ovariectomized rats with alpha-methyl-p-tyrosine (alpha MT) caused a 91% reduction in the concentration of DA in portal plasma. Infusion of DA (0.4 microgram/min/kg BW) into a jugular vein of rats pretreated with alpha MT restored the DA concentration in portal plasma to that seen in untreated rats. Injection of morphine sulfate elicited a marked increase in the concentration of PRL in plasma. Infusion of DA at rates of 0.4 and 0.8 microgram/min/kg BW suppressed by 52 and 75%, respectively, the secretion of PRL after morphine treatment. Infusion of DA had no effect on the release of PRL induced by intracerebroventricularly administered beta-endorphin. Although treatment of rats with alpha MT caused release of PRL that was similar to that seen in rats treated with morphine, infusions of DA at 0.4 and 0.8 microgram/min/kg BW into alpha MT-treated, ovariectomized rats suppressed the secretion of PRL by 89 and 96%, respectively. Thus, after morphine treatment, the decreased release of DA into portal blood is not in itself sufficient to account for the increase seen in the secretion of PRL. It is suggested that morphine and opiate-like peptides induce the release of a hypothalamic substance(s) that stimulates PRL release.     

Endogenous GABA receptor ligands in hypophysial portal blood

The concentrations of endogenous gamma-aminobutyric acid (GABA) receptor ligands were measured in hypophysial portal plasma by a sensitive and specific radioreceptor assay. Portal plasma contained higher concentrations of GABA receptor ligands than peripheral controls, although this increase was not due to authentic GABA but an unknown low-affinity ligand. Evidence is presented to support the hypothesis that the dipeptide homocarnosine may represent at least a part of this unknown. Electrical stimulation of median eminence evoked a massive (7.8-fold) increase in the rate of release of GABA (but not the low-affinity component) into portal blood. These results suggest that GABA and possibly other low-affinity ligands such as homocarnosine could serve as factors involved in the central control of anterior pituitary function.     

Release of prolactin is independent of the secretion of thyrotrophin-releasing hormone into hypophysial portal blood of sheep

In order to determine whether pituitary prolactin release was directly related to the secretion of TRH into hypophysial portal blood, serial portal and jugular venous blood samples were collected from seven lactating and three non-lactating ewes. In another experiment, samples were collected from five ovariectomized ewes while being exposed to an audio-visual stress and then later administered with chlorpromazine. Secretion of TRH was pulsatile in all ewes and independent of prolactin secretion; TRH pulses coincided with significant increases in prolactin secretion in only 15% of cases and only 29% of prolactin pulses were associated with TRH pulses. Sixty-seven per cent of suckling bouts were associated with increases in prolactin secretion, but only 22% of these were associated with significant increases in TRH secretion. Chlorpromazine increased prolactin levels fourfold but did not affect portal concentrations of TRH. Audio-visual stress was not a reliable method of causing prolactin release in this model. Mean portal concentrations of TRH and jugular concentrations of prolactin were not significantly correlated. These results show that hypothalamic TRH and pituitary prolactin are secreted independently in the sheep, implying that increases in prolactin release caused by suckling or chlorpromazine are not the direct result of increased TRH secretion.     

Oxytocin and vasopressin in rat hypophysial portal blood: experimental studies in normal and Brattleboro rats

Oxytocin (OT) and vasopressin (VP) were measured by radioimmunoassay in hypophysial portal and peripheral blood from male Wistar rats and heterozygous and homozygous Brattleboro rats anaesthetized with urethane. In Wistar rats the concentrations of OT and VP were about 50 times greater than the concentrations in peripheral blood, whether or not the pituitary gland was left in situ during collection, and also considerably greater than the reported concentrations of the peptides in the cerebrospinal fluid. The release of both peptides was increased significantly by a lesion of the supraoptico-hypophysial tract that led to diabetes insipidus, but which left intact the external layer of the median eminence (ME). Concentrations of VP were undetectable in plasma from homozygous Brattleboro rats, but the portal plasma concentrations of VP in heterozygous Brattleboro rats were not significantly lower than in Wistar rats. The concentrations of OT in portal plasma from both types of Brattleboro rat were significantly higher than in Wistar rats. The output of VP and OT into hypophysial portal blood of Wistar rats was not significantly affected by electrical stimulation of the suprachiasmatic, supraoptic or paraventricular nuclei or the ME using two types of stimuli, one of which produced an increase in peripheral plasma concentrations of VP and OT in intact rats and a significant increase in the release of LH-releasing hormone into hypophysial portal blood. The output of VP and OT into portal blood was also not significantly affected by either adrenalectomy with or without injection of dexamethasone or the injection of either the 5-hydroxytryptamine (5-HT) synthesis blocker, parachlorophenylalanine, or the 5-HT uptake blockers, alaproclate or zimelidine. These results show that large amounts of OT as well as VP are released into hypophysial portal blood from fibres of the hypothalamo-neurohypophysial system that terminate in the external layer of the ME. Although distinct from the fibres that terminate in the pars nervosa (PN), the findings in Brattleboro rats show that the VP fibres of the ME system originate in neurones with a genomic mechanism for VP synthesis similar to that of the VP neurones that project to the PN. The lack of effect of adrenalectomy and the administration of 5-HT synthesis and uptake blockers must be interpreted with caution since the results obtained with electrical stimulation suggest that when the pituitary stalk is cut the release of OT and VP into portal blood approaches a maximum and may therefore be difficult to alter by experimental manipulation.(ABSTRACT TRUNCATED AT 400 WORDS)     

The corticotropin-releasing factor release in rat hypophysial portal blood is mediated by brain catecholamines

In order to study the involvement of the hypothalamic corticotropin-releasing factor (CRF) in catecholamine-induced adrenocorticotropin (ACTH) secretion, we have measured CRF levels in rat hypophysial portal blood (HPB) after the pharmacological destruction of the ventral noradrenergic bundle (VNAB), using 6-hydroxydopamine (6-OHDA) stereotaxically injected into the VNAB. CRF levels in HPB were measured by radioimmunoassay, and the effects of 6-OHDA injection were controlled by the determination of catecholamine concentrations in the total hypothalamus. VNAB lesions induced a dramatic decrease in norepinephrine and epinephrine hypothalamic concentration. The CRF levels in HPB were also significantly reduced. These results suggest that central catecholamines exert a direct stimulatory control on the CRF release and play a major role in stress-induced ACTH secretion.     

Direct measurement of blood pressure within the long hypophysial portal blood vessels

The microvascular pressures that perfuse the anterior pituitary gland with blood were not known. We now report the direct measurement of these pressures in the urethane-anesthetized rat. The infundibular stalk and ventral surface of the anterior pituitary gland were surgically exposed via a parapharyngeal approach and a micropressure transducer inserted into the lumen of hypophysial portal vessels under direct microscopic observation. A Weiderhielm-type servo-controlled pressure system was used to record the pressures. Continuous pressure recordings up to 30 min in duration were made in long hypophysial portal vessels ranging in diameter from 10 to 50 micron in adult, female Sprague-Dawley rats. The mean pressure recorded from these vessels was 4.0 cm H2O (2.7 mm Hg). A small increase in systemic pressure produced by a rapid saline infusion into a cannulated femoral vein resulted in a mirrored but much greater magnitude increase in pressure to the hypophysial portal vessels. This finding suggests that pressure within the portal vessels is in some instances closely coupled to systemic blood pressure. The low pressures recorded in hypophysial portal vessels correlate well with pressures measured in the hepatic portal vasculature. The porosity of fenestrated capillaries surrounding anterior pituitary cells is hemodynamically essential, since the low hydrostatic pressures alone would be inappropriate for rapid and thorough exchange.     

Presence of corticotropin releasing factor-like immunoreactivity in hypophysial portal blood

Hypophysial protal blood was collected from pentobarbital anesthetized male rats, and the content of corticotropin releasing factor-like immunoreactivity (CRF-LI) was measured using an RIA developed for synthetic ovine CRF. The concentration of CRF-LI in portal plasma was 104.9 +/- 9.7 pM. This concentration is in the range which has been shown to stimulate ACTH and beta-endorphin secretion in vitro. These data support the hypothesis that a molecule with the same or similar structure as synthetic ovine CRF is a physiologically significant hypothalamic releasing factor.     

Dopamine in hypophysial portal plasma of the rat during the estrous cycle and throughout pregnancy

Catecholamine levels in hypophysial portal plasma were determined in pregnant and non-pregnant female rats as well as in intact and castrated male rats, using a radioenzymatic assay for the simultaneous determination of dopamine, norepinephrine, and epinephrine in 50 mul of plasma. Portal and arterial blood were collected from anesthetized rats at 7 mul/min for 60 min. During the collection, blood was kept at 0 C, a temperature at which endogenous catecholamines were relatively stable. Dopamine was present in high concentrations in hypophysial portal plasma thorughout pregnancy, attaining a level near 20 ng/ml on the 20th day of gestation. Dopamine levels in arterial plasma from the same rats were low or undetectable (0.4--0.8 ng/ml1. Norepinephrine and epinephrine was undetectable (less than 0.6 ng/ml) in portal as well as arterial plasma from these rats. The major catecholamine in extracts of the hypothalamus from pregnant rats was norepinephrine, whereas that in the posterior pituitary was dopamine. Dopamine levels in portal plasma collected during proestrus, estrus, diestrus 1, and diestrus 2, were 1.32 +/- 0.21 (mean +/- SE), 3.87 +/- 0.96, 3.11 +/- 0.73, and 2.3 +/- 0.45, respectively. Dopamine in portal plasma from intact and from castrated male rats was approximately 0.6 ng/ml. Norepinephrine and epinephrine were not detectable in either portal or arterial plasma from these animals. It is concluded 1) that dopamine is secreted into hypophysial portal blood in significant quantities during pregnancy, 2) that hypothalamic secretion of dopamine in cyclic rats is greatest during the day of estrus and early diestrus and at least on the day of proestrus, and 3) that these findings support the view that dopamine of hypothalamic origin may have an important role in the regulation of anterior pituitary function.     

Levels of dopamine and thyrotrophin-releasing hormone in hypophysial stalk blood during an oestrogen-stimulated surge of prolactin in the ovariectomized rat

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 micrograms 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.