Transient increase in prolactin secretion following hypothalamo-pituitary disconnection in ewes during anoestrus and the breeding season

Hypothalamic control of prolactin secretion was studied in ovariectomized ewes by comparing the effects of hypothalamo-pituitary disconnection (HPD) and sham-operation (sham-HPD). HPD caused a two-fold increase in plasma prolactin concentrations on days 1 and 7 following surgery during anoestrus and a tenfold increase during the breeding season. Thereafter, concentrations gradually declined to be similar to those in sham-HPD ewes by day 43 (breeding season) and day 145 (anoestrus). The maximum plasma prolactin response to HPD was similar during the two seasons (anoestrus: 128 +/- 15 bs breeding season: 118 +/- 13 micrograms/l). Sham-HPD had no effect on plasma prolactin concentrations. Prolactin pulse frequency was not affected by HPD, but increases in plasma prolactin concentrations were associated with increases in pulse amplitude. At the time of the normal anoestrus, plasma prolactin concentrations rose in both the HPD and sham-HPD ewes, raising the question of extra-hypothalamic regulation of seasonal changes in prolactin secretion. Plasma LH and FSH concentrations became undetectable in HPD ewes but were unaltered in sham-HPD ewes. We conclude that hypothalamic inhibition of pituitary prolactin secretion in the sheep can be demonstrated by HPD but that this effect is not sustained. This transience may indicate the additional requirement of hypothalamic-releasing factors in the control of prolactin release. In addition, the surgically isolated ovine pituitary of the HPD animal has an inherent pulsatile secretion of prolactin.     

Decreased prolactin responsiveness to thyrotrophin-releasing hormone and metoclopramide in hyperthyroidism

To investigate whether prolactin (Prl) responsiveness to thyrotrophin-releasing hormone (TRH) differs in thyrotoxic and normal individuals, serum Prl was determined before and after iv injection of 200 micrograms TRH in 10 patients with untreated thyrotoxicosis and also in 9 normal subjects. Both the maximal Prl increment after TRH and the total Prl response, represented by the Prl incremental area, were significantly larger in the normal subjects compared with the thyrotoxic (max Prl increment 56 +/- 11 vs 15 +/- 3 ng/ml, P less than 0.001; Prl incremental area 3071 +/- 522 vs 579 +/- 171, P less than 0.001; mean +/- SEM). The maximal Prl increase after 15 mg oral metoclopramide (MET) was also significantly larger in the normal (125 +/- 13 ng/ml) than in the thyrotoxic subjects (60 +/- 13 ng/ml, P less than 0.01). When 200 micrograms TRH was injected iv 90 min after oral administration of 15 mg MET, an additional Prl increase was observed in normal individuals (21 +/- 6 ng/ml, P less than 0.01). In thyrotoxic patients, however, iv TRH failed to induce a significant increase in Prl after oral priming with MET (0 +/- 3 ng/ml). When 7 thyrotoxic patients, made euthyroid by 125I-treatment, were investigated according to the same protocol as the one mentioned above, they displayed normal Prl responses to iv TRH and to oral MET. Furthermore, they showed a significant Prl response to iv TRH after oral priming with MET (20 +/- 8 ng/ml, P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

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.     

Effect of photoperiod and morphine on plasma prolactin concentrations and thyrotropin-releasing hormone secretion in the ewe

This study investigated the effects of daily photoperiod length and morphine on thyrotropin-releasing hormone (TRH) and prolactin secretion in sheep. Two groups of adult ewes were kept under either 16 h L:8 h D or 8 h L:16 h D photoperiods for approximately 60 days. Then the photoperiods were reversed and approximately 60 days later push-pull cannulae were implanted into the hypothalamic stalk-median eminence. After a 7-day recovery period, the ewes were subjected to hypothalamic perfusion and blood was collected at 15-min intervals from the jugular vein. Perfusates also were collected into 15-min fractions. The first 3 h of perfusion served as an equilibrium period. During the next 2 h, saline was infused into one jugular vein. This was followed by a 2-hour morphine infusion (1 mg.kg-1.h-1). Data from 13 ewes were analyzed for effect of photoperiod and drug on TRH concentrations in the perfusates and prolactin in the serum. Prolactin was significantly (p less than 0.01) higher under 16 h L:8 h D than 8 h L:16 h D and was greatly increased (p less than 0.001) by morphine infusion. TRH only tended to be higher (p = 0.05) under 16 h L:8 h D than under 8 h L:16 h D, but morphine infusion induced a rapid and highly significant (p less than 0.01) increase in TRH secretion. There were no photoperiod and drug interactions for either TRH or prolactin.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of modifying gonadotrophin-releasing hormone input before and after the oestrogen-induced LH surge in ovariectomized ewes with hypothalamo-pituitary disconnection

The patterns of gonadotrophin-releasing hormone (GnRH) input to the pituitary gland that affect the expression of a positive-feedback event by oestrogen on LH secretion were investigated in ovariectomized ewes with hypothalamo-pituitary disconnection (HPD). In experiment 1, ovariectomized HPD ewes were given hourly i.v. pulses of 250 ng GnRH and an i.m. injection of 50 micrograms oestradiol benzoate (OB). The ewes were given a bolus pulse of 2.25 micrograms GnRH 16 h after injection of OB, followed by half-hourly pulses of 250 ng GnRH for 14 h (treatment A). The LH surge response was significantly (P less than 0.05) greater in these ewes compared with that in ewes given a continuous infusion of GnRH (250 ng/h) after the OB injection, followed by a continuous infusion of 500 ng GnRH/h after the bolus pulse of GnRH (treatment B). When no GnRH was administered after the OB injection, except for the bolus pulse of GnRH (treatment C), the surge response was significantly (P less than 0.05) reduced compared with that in treatment A, and was reduced compared with treatment B. These data suggest that GnRH pulses are important in the generation of the OB-induced LH surge, but that a baseline secretory component can prime the pituitary to some extent. In experiment 2, a doubling of the continuous infusion dose of GnRH used in treatment B to 500 ng/h before the bolus pulse of GnRH and to 1 micrograms/h afterwards (treatment D) gave a similar response compared with treatment A, suggesting that if the baseline input of GnRH is of sufficient magnitude, it can overcome the lack of pulsatile input. In experiment 3, halving the GnRH pulse amplitude used in treatment A from 250 to 125 ng (treatment E) did not reduce the LH surge response, implying that when the GnRH input is in a pulsatile mode, the amplitude of GnRH pulses is less important than the pulsatile nature per se. In experiment 4, removal of GnRH input after the bolus pulse of GnRH (treatment F) significantly (P less than 0.05) reduced the surge response compared with when pulses were maintained (treatment A), indicating that GnRH input is still required once the LH surge has been initiated. Collectively, these experiments show that several forms of GnRH delivery, both pulsatile and baseline, can result in the full expression of a positive-feedback response in ovariectomized ewes treated with oestrogen.     

Physiological control of growth hormone secretion by thyrotrophin-releasing hormone in the domestic fowl

Immature cockerels (4- to 5-weeks old) were passively immunized, with antiserum raised in sheep, against thyrotrophin-releasing hormone (TRH). The administration of TRH antiserum (anti-TRH) at doses of 0.5, 1.0 or 2.0 ml/kg lowered, within 1 h, the basal concentration of plasma GH for at least 24 h. The administration of normal sheep serum had no significant effect on the GH concentration in control birds. Although the GH response to TRH (1.0 or 10.0 micrograms/kg) was not impaired in birds treated 1 h previously with anti-TRH, prior incubation (at 39 degrees C for 1 h) of TRH (20 micrograms/ml) with an equal volume of anti-TRH completely suppressed the stimulatory effect of TRH (10 micrograms/kg) on GH secretion in vivo. These results suggest that TRH is physiologically involved in the hypothalamic control of GH secretion in the domestic fowl.     

Role of hypothalamic catecholamines in the regulation of luteinizing hormone and prolactin secretion in the ewe during seasonal anestrus

Separate studies with ewes have shown that catecholamines play an inhibitory role in the control of LH secretion during anestrus, and that there are structures in the lateral retrochiasmatic area (L-RCh), which could be involved in the regulation of gonadotrophin secretion. These observations have led to the hypothesis that the catecholaminergic structures in the L-RCh mediate the inhibition of pulsatile LH secretion by estradiol in the anestrous ewe. We tested this hypothesis by injecting 6-hydroxydopamine (6OH-DA) into the L-RCh of ovariectomized ewes during the anestrous season, and comparing the secretion of LH and prolactin in these animals with that in sham (injected with vehicle) and control (no injection) animals, in the presence and absence of exogenous estradiol. Finally, the effectiveness of the toxin was assessed by immunocytochemical techniques. When the ewes were treated with estradiol, LH pulse frequency was significantly lower in the controls (mean 1.1 pulses/4 h) and shams (0 pulses/4 h) than in the ewes treated with 6OH-DA (3.1 pulses/4 h). When the estradiol implants were removed, the frequencies increased to 5.1 pulses/4 h for the controls and 5.7 pulses/4 h in the ewes treated with 6OH-DA. These were not significantly different. Plasma prolactin levels were significantly reduced by 6OH-DA treatment. The 6OH-DA ewes recovered their response to estradiol by 14 weeks after the injection. The anatomical study at the end of the experiment revealed a difference between treated and control ewes of only 15% in the numbers of dopaminergic cells in the L-RCh.(ABSTRACT TRUNCATED AT 250 WORDS)     

Short-loop inhibition of thyrotrophin-releasing hormone-induced growth hormone secretion in fowl

The i.c.v. administration of 0.1 or 10 micrograms ovine (o)GH to 12- to 16-week-old hypothyroid chickens of a sex-linked dwarf (SLD) strain suppressed the basal plasma GH concentrations, measured 24 h afterwards. The GH response of the oGH-injected SLDs to TRH was suppressed, in a dose-related way, in comparison with that induced by TRH in birds given control injections (10 micrograms) of bovine serum albumin (BSA). Basal circulating concentrations of GH in euthyroid K strain birds of the same age were even lower than in the SLDs following injection of 10 micrograms oGH, and were not further reduced by oGH administration. The GH response to TRH in the K strain birds injected i.c.v. with 0.1 or 10 micrograms oGH was, nevertheless, suppressed in comparison with the BSA-injected K strain controls. The i.c.v. administration of oGH also suppressed circulating concentrations of LH and the LH response to TRH in the K strain birds. Twenty-four hours after i.c.v. administration of oGH (10 micrograms), the somatostatin (SRIF) content in the medial basal hypothalamus of 8-week-old euthyroid cockerels was greater than that in BSA (10 micrograms)-injected controls. At the same time, the binding of [3H]3-methyl-histidine2-TRH to the pituitary caudal and cephalic lobes of GH-injected birds was less than that in the controls. These results suggest that GH regulation in avian species is partly mediated by an inhibitory short-loop mechanism (mediated by hypothalamic SRIF and a down-regulation of pituitary TRH-binding sites) that suppresses basal and secretagogue-induced GH release.     

Anterior pituitary response to thyrotrophin-releasing hormone during open-heart surgery.

The anterior pituitary response to thyrotrophin-releasing hormone has been studied in 20 patients submitted to elective open-heart surgical procedures, and in six control patients submitted to closed mitral valvotomy. Standard non-pulsatile normothermic perfusion was used in all the open-heart cases. 400 microgram thyrotropin-releasing hormone was administered by intravenous injection during bypass, at 30 min post-bypass, and at 60 min post-bypass. The same dose (400 microgram) was given during closed mitral valvotomy (Control Group). Thyrotrophin-releasing hormone administration during bypass failed to produce a normal response from the anterior pituitary, in contrast to the normal response pattern seen in the control group (P less than 0.01). Thyrotrophin-releasing hormone given after the period of bypass produced responses within the normal range in the majority of patients. These results suggest that anterior pituitary hypofunction may exist during the period of extracorporeal circulation using non-pulsatile perfusion and that recovery of pituitary function is evident within the first hour post-extracorporeal circulation.     

Regulation of prolactin secretion during suckling: involvement of the hypothalamo-pituitary GABAergic system

Previous results demonstrated that GABA exerts a dual control on PRL secretion, one excitatory mediated in part by the impairment of the tubero-infundibular dopaminergic (TIDA) system function, the other inhibitory occurring at the level of the anterior pituitary (AP), where 3H-GABA and 3H-Muscimol (3H-M) recognition sites have been described. This report provides evidence for a physiological role of the tubero-infundibular GABAergic system (TI-GA-BA) on PRL secretion in the rat. In lactating rats separated for 4 h from their pups reinstitution of suckling for different periods resulted in an increase either in glutamic acid decarboxylase (GAD) activity in the mediobasal hypothalamus (MBH) or in AP-GABA content. Dynamic changes of the GABAergic function in the MBH-AP system seemed to have a certain degree of specificity because suckling did not affect GAD activity in the caudate nucleus. In lactating rats 2, 4, 8 and 24 h after removal of the offsprings AP-GABA concentrations and plasma PRL titers significantly decreased with respect to values present in rats never separated from their pups. Since it has been demonstrated that the PRL lowering effect of GABA is a receptor-mediated event, we have investigated the plasticity of AP-GABA receptors during suckling. The inhibitory action of GABA seems to be mediated mainly by the activation of the high affinity binding sites. This proposition is supported by the fact that in lactating rats, where only the high affinity receptor population is present, M was still able to decrease significantly plasma PRL concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)     

Seasonal influence on thyroidal response to thyrotrophin-releasing hormone in cattle and buffaloes

The effect of thyrotrophin-releasing hormone (TRH) on the circulating plasma levels of tri-iodothyronine (T3) and thyroxine (T4) was determined in the same group of animals (four cattle and four Murrah buffaloes) during hot dry (HD), hot humid (HH) and cold environmental conditions. Plasma T3 and T4 concentrations were measured during 2 h before and up to 12 h after the administration of TRH (200 micrograms i.v.). In the preinjection period in both cattle and buffaloes T3 levels were significantly lower in HH conditions. No significant difference in basal (preinjection) T3 levels was observed during HD and cold seasons in cattle. The highest T3 levels were obtained in buffaloes during HD season with intermediate values during the cold months. Plasma T4 levels in these animals were reversed during HD and HH months. In both cattle and buffaloes there was a biphasic response of T3 and T4 to TRH treatment and this varied with time and in size. The season significantly affected the T3 response to TRH in cattle and buffaloes but the T4 response differed in the two species. The ratio of T4/T3 was higher during HH condition compared with other seasons in both cattle and buffaloes. The climate significantly affected the thyroidal response to TRH.     

Enhanced plasma thyrotrophin response to thyrotrophin-releasing hormone following oestradiol administration in man

The plasma TSH response to 100 μg of TRH i.v. was studied in six normal women and in seven men before and after the administration of 10.0 mg of oestradiol valerate for 5 consecutive days. Before oestrogen treatment the females appeared more responsive than males. After treatment the response remained unchanged in females while appearing enhanced in males. Oestrogen treatment did not modify either the basal plasma TSH concentration or the serum thyroxine, the labelled tri-iodothyronine uptake and the calculated free thyroxine index. It is concluded that oestrogens play a role in modulating the secretion of TSH from the human pituitary.     

Prolactin short-loop feedback and prolactin inhibition of luteinizing hormone secretion during the breeding season and seasonal anoestrus in the ewe

In seasonally breeding mammals, plasma prolactin (PRL) concentrations vary on an annual basis with levels high in summer and low in winter. In this study of the ewe, we determined, first, whether PRL secretion is regulated by short-loop feedback and, second, whether the high summer levels of PRL are due to a change in sensitivity or loss of this feedback loop. Because the high summer levels of PRL coincide with the period of seasonal anoestrus in the ewe and could therefore be involved in the seasonal suppression of gonadotrophins, the effects of intracerebroventricular PRL on pulsatile LH secretion were also determined. Ovary intact ewes received intracerebroventricular injections of ovine PRL (oPRL; 50 micrograms) or anti-PRL serum. From 3 to 13 h after central administration of oPRL, plasma PRL concentrations were significantly reduced compared with the vehicle-treated controls. In contrast, following injection of anti-PRL serum, plasma PRL levels increased significantly. To determine whether there was a seasonal change in sensitivity to PRL feedback, a series of experiments were conducted in July and November when PRL concentrations are high and low, respectively. At each time of year, ovariectomized oestradiol-implanted ewes were injected intracerebroventricularly with 10 and 50 micrograms oPRL with control animals receiving the vehicle. At both times of year there was clear evidence of PRL short-loop feedback with no indication that sensitivity was reduced in the July trial. Luteinizing hormone (LH) pulse frequency, pulse amplitude and mean LH were not affected by intracerebroventricular oPRL at either time of year.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pineal melatonin secretion drives the reproductive response to daylength in the ewe

This study was conducted to determine whether the pineal indoleamine melatonin mediates the effects of photoperiod on the capacity of estradiol to inhibit LH secretion in the ewe. Patterns of serum melatonin were characterized in pineal-intact ovariectomized ewes treated with sc Silastic estradiol implants and exposed to 90-day alternations between long and short photoperiods. High fluctuating levels of serum melatonin were found during the night, with the duration of elevated serum levels corresponding to the length of the dark period. Transfer from long to short photoperiods caused a rapid change in the duration of nightly melatonin secretion and reduced the negative feedback potency of estradiol upon LH secretion during the natural anestrous season. In pinealectomized ewes, the night-time rise of melatonin was absent, and transfer from long to short days failed to reverse the capacity of estradiol to inhibit LH secretion during anestrus. Nightly infusions of melatonin restored patterns of this indoleamine similar to those observed in pineal-intact ewes exposed to the 90-day alternation between long and short days. The melatonin infusions also restored the reproductive response to the inductive photoperiod: in every ewe, the negative feedback effects of estradiol upon LH secretion were diminished after transfer from long to short days. The amplitude and latency of this escape matched those of pineal-intact animals. We conclude that the pineal mediates the reproductive response of the ewe to inductive photoperiods through its daily rhythm of melatonin secretion.