An experiment was carried out to investigate the effect of a range of estradiol (E2) doses (0.1-6.5 micrograms/g body wt/day) on vitamin D metabolism and the plasma levels of growth hormone (GH) and prolactin (PRL) in the growing chick. Doses of 0.5-0.7 microgram/g E2, which are insufficient to raise the plasma calcium level, did induce an increase in growth rate, an increase in 25-hydroxyvitamin D 1 alpha-hydroxylase (1-hydroxylase) and 24-hydroxylase activities, and an increase in plasma GH level. These parameters leveled off or fell over the dose range 1-2 micrograms/g E2 but there was evidence of a second peak in 1-hydroxylase activity at 6 micrograms/g E2. At this high dose rate, the plasma Ca level rose to 8 mM, as it does in the laying hen; 24-hydroxylase activity, growth rate, and plasma GH and plasma PRL levels all decreased. It was concluded that the dose response to estrogen in the growing chick is not linear and, in the case of 1-hydroxylase activity, may even be biphasic.

The basal release of prolactin from cockerel anterior pituitary glands in vitro declined between 1 and 7 weeks of age, to a level less than that released by pituitary glands from 18 week old (adult) cockerels and hens. Basal growth hormone (GH) release increased between 1 and 7 weeks of age but had declined in adults to a level similar to that released from 4 weeks old cockerels. The responsiveness of the pituitary gland to hypothalamic stimulation, using hypothalami from 8 week old broiler fowl, was also age-related. Prolactin release was considerably higher from pituitaries of 1 week old cockerels compared to the other age groups. Stimulation of GH release by the hypothalamus was higher from pituitaries of both 1 and 7 week old cockerels compared to the other groups of birds. The increase in release of prolactin following incubation with thyrotrophin releasing hormone (TRH) declined between 1 and 7 weeks, but increased slightly in adult birds, whereas the increase in release of GH following TRH was higher from pituitaries of both 1 and 7 week old cockerels. Hypothalamic prolactin (Prl) releasing activity, measured as the ability of the hypothalamus to stimulate hormone release from 8 week old broiler fowl anterior pituitary glands, declined with the age of the donor cockerels. The hypothalami from adult hens secreted significantly more Prl releasing activity than did adult cockerel hypothalami. The secretion of GH releasing activity decreased markedly with the age of the donor bird. These results suggest that maturational patterns of hormone secretion in fowl are partly due to changes in autonomous hormone release, to changing patterns of hypothalamic activity and to differences in pituitary responsiveness to provocative stimuli.

The plasma concentration of prolactin in immature cockerels was increased between 10 and 40 min after the intravenous administration of prostaglandin (PG) F2 alpha (200 micrograms/kg body weight). Lower doses had no effect on plasma prolactin concentration. The addition of PGF2 alpha (10(-9) to 10(-6) M) to incubation media had no effect on the basal release of pituitary prolactin but reduced the release of prolactin from pituitary-hypothalamus co-incubations. The addition of noradrenaline (10(-7) M), serotonin (10(-7) M), acetylcholine (10(-6) M) or histamine (10(-6) M) to the co-incubation increased the hypothalamus-induced prolactin release, although these effects were not observed in the presence of 10(-7) M PGF2 alpha. The in vitro release of pituitary prolactin was increased by adding chicken hypothalamic extract in the presence or absence of PGF2 alpha. These results suggest a dual effect on PGF2 alpha of prolactin secretion in the fowl; its stimulation in vivo may result from a peripheral action.

Young cockerels injected 24 h earlier with 0.9% saline,para-chorophenylalanine (pCPA, brain serotonin depletor) or alpha-methylpara-tyrosine (AMPT, brain catecholamine depletor) were deprived of access to water for 24 h. Plasma prolactin concentrations were markedly elevated by water deprivation and returned to normal on rehydration. pCPA, but not AMPT, significantly reduced the increase in prolactin. Concentrations of growth hormone were not affected by water deprivation. Brain serotonin concentrations were reduced by treatment with pCPA. Groups of cockerels were maintained under normal conditions or without access to drinking water for 12 h or 24h. Some were injected with the monoamine oxidase inhibitor pargyline, which increased the prolactin and decreased the growth hormone concentration in the plasma of the hydrated birds. The inhibitory effect of pargyline on growth hormone was augmented following water deprivation. Serotonin levels were not significantly affected by water deprivation but turnover (defined as accumulation of serotonin after pargyline treatment) was increased in the hypothalamus but not in remaining tissue. Injecting 30% saline solution intravenously markedly increased plasma prolactin whilst growth hormone concentrations were decreased. Serotonin turnover was increased in the hypothalamus but not in other brain regions. The results show that secretion of prolactin and growth hormone by the pituitary gland during osmotic imbalance in the fowl may be mediated by changes in hypothalamic scrotonin turnover.

Pituitary glands and hypothalami from broiler fowl heads were incubated alone or together with histamine, gamma-aminobutyric acid (GABA) or acetylcholine (ACh) as well as with catecholamines or neurotransmitter antagonists. Histamine and ACh stimulated, whereas GABA inhibited, the hypothalamus-induced release of prolactin, responses blocked by their specific antagonists. The dopamine antagonist pimozide, but not adrenergic (both alpha and beta), serotoninergic or cholinergic antagonists, prevented the actions of histamine and GABA. None of the antagonists except the cholinergic blocker, atropine, affected ACh-induced release of prolactin. Neither histamine nor ACh prevented inhibition of prolactin release by dopamine or stimulation of prolactin release by noradrenaline. GABA did not affect the response to noradrenaline. Furthermore, histamine, GABA and ACh had no effects on thyrotrophin releasing hormone-stimulated release of prolactin directly at the pituitary level. These results suggest that histamine and GABA affect prolactin release from chicken pituitaries in vivo by modifying the activity of the dopaminergic system. Acetylcholine may stimulate the secretion of prolactin releasing factor from the hypothalamus.

Ten-day old chicks were divided into 6 groups which were fed a low phosphorus (P) diet for periods of 0, 0.5, 1, 3, 7 or 11 days before killing at 3 weeks old. Labelled calcium (47Ca) was injected intraperitoneally into some birds 18 hours before killing. A marked fall in growth rate, plasma phosphorus level, plasma growth hormone level and renal 24-hydroxylase activity levels had occurred by 12 hours after the experimental diet had started. After one day on the diet, the rate of duodenal Ca absorption had risen and continued to rise up to the 11th day. During this period, the renal 25-hydroxyvitamin D-1-hydroxylase activity rose slightly while the 24-hydroxylase activity rose towards the control level. At 24 hours, the 47Ca level in the bone was markedly lower than in the control group and remained low. It was concluded that the first adaptive response of the chick to dietary P insufficiency was to suppress growth. Subsequent adaptive responses were to increase the rate of Ca and P absorption from the gut and mobilisation from the bone but despite these measures, the growth rate only recovered slightly and the plasma P level continued to fall.

Fowl anterior pituitary glands were bisected and each half was pretreated in either Medium 199 or medium containing EGTA to deplete endogenous calcium (Ca2+) stores, after which they were incubated in Medium 199, or Ca2+-free medium, containing prolactin release-stimulating agents and verapamil, a Ca2+ channel blocker. High K+ concentrations, hypothalamic extract, synthetic thyrotrophin-releasing hormone (TRH) and dibutyryl cyclic AMP (dbcAMP) all stimulated release of prolactin from control (non EGTA-treated) hemianterior pituitary glands. The effects of TRH and dbcAMP were not additive, but the response to submaximal concentrations of TRH was augmented by theophylline, a phosphodiesterase inhibitor. Reduction of Ca2+ availability with EGTA or verapamil reduced basal release of prolactin, prevented the prolactin-stimulating effects of high K+ concentrations and TRH, and markedly attenuated responses to hypothalamic extract and dbcAMP, EGTA being more effective than verapamil. Increasing the Ca2+ concentration of the medium did not augment basal or stimulated release of prolactin. These results suggest that both Ca2+ and cyclic AMP may act as intracellular mediators in the release of prolactin. Both basal and stimulated release of prolactin depend upon the presence of Ca2+. Although influx from the medium may be the major source of Ca2+, endogenous stores of Ca2+, perhaps mobilized by dbcAMP, may be able to maintain some release of prolactin. The prolactin-stimulating effects of TRH may be mediated by cyclic AMP.

Anterior pituitary glands from broiler fowl were preincubated for 24 h in either medium 199 only or medium containing estradiol 17β, following which they were incubated in medium containing thyrotrophin releasing hormone (TRH), vasoactive intestinal polypeptide (VIP) or substance P (SP), alone or with the dopamine agonist, apomorphine. Estradiol priming stimulated release of prolactin and enhanced apomorphine-inhibition of prolactin release. TRH stimulated prolactin release, an effect reversed by apomorphine, and priming with estradiol potentiated both effects. VIP stimulated prolactin to a lesser degree and again this was inhibited by apomorphine and potentiated by estradiol. SP had little effect on the nonsteroid-primed pituitary, but stimulated release of prolactin after estradiol treatment, though less effectively than TRH or VIP.

Anterior pituitary glands from broiler fowl were incubated by themselves, with hypothalamic tissue or with thyrotrophin releasing hormone (TRH) in medium containing dopamine and its antagonist pimozide. The presence of hypothalamic tissue or TRH resulted in a stimulation of release of prolactin. Neither dopamine nor pimozide affected prolactin release directly from the pituitary gland. Dopamine inhibited the release of prolactin stimulated by hypothalamic tissue or TRH, in a concentration-dependent fashion. Pimozide diminished the response to dopamine. After pituitary glands were preincubated for 20 h in medium containing oestradiol-17 beta, the basal release of prolactin was enhanced as was the response to TRH. Both basal and TRH-stimulated release of prolactin from the oestrogen-primed pituitary glands was inhibited by dopamine, an effect blocked by pimozide. Hypothalami from broiler fowl were incubated for up to 8 h in medium containing dopaminergic drugs and pituitary glands were incubated in this medium, alone or with pimozide. As indicated by the prolactin released by the pituitary glands, the hypothalami appeared to secrete prolactin-releasing activity in a time-related fashion. Dopaminergic activity was also present in the hypothalami, since pimozide enhanced the prolactin-releasing activity of the medium. Dopamine apparently inhibited and pimozide stimulated the secretion of releasing activity from the hypothalamus. These results suggest that dopamine inhibits release of prolactin directly from the pituitary gland only when prolactin secretion is high. The hypothalamus secretes at least two factors regulating prolactin secretion, a prolactin-releasing factor and a dopaminergic prolactin-inhibiting factor. Dopamine may also play an inhibitory role in the regulation of secretion of the prolactin-releasing factor.

1. Pituitary glands and hypothalami from broiler fowl heads were incubated alone or together with dopamine, noradrenaline or monoaminergic drugs (apomorphine, pimozide, phentolamine, isoproterenol and propranolol).

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.

When pigeons were injected with an extract of chicken hypothalamus, the increase in weight of the crop sac showed that prolactin secretion was stimulated. Crop sac weight was no higher after 16 days of injection that it was after 8 days. Pituitary gland content of prolactin and growth hormone was elevated at 8 days, but showed a relative decrease after a further 8 days of injections. Prolactin and growth hormone releasor activity in the hypothalamus was investigated in vitro. A decline during treatment with hypothalamic extract was seen, and by 8 days no releasor activity could be detected. Injection with ovine prolactin also reduced prolactin releasor activity in the hypothalamus. The results suggest that the secretion of prolactin and possibly also of growth hormone is controlled by a “short loop” negative feedback mechanism which acts on hypothalamic hypophysiotrophic hormones.

Anterior pituitary glands from broiler fowl were incubated alone or with hypothalamic tissue in medium containing either serotonin or serotoninergic drugs, acetylcholine or cholinergic drugs, and the release of prolactin (Prl) and growth hormone (GH) measured by homologous radioimmunoassays. The neurotransmitters and drugs affected the release of hormones from the pituitary gland only when hypothalamic tissue was also present. Serotonin and its agonist quipazine stimulated the release of Prl and inhibited release of GH in a concentration-related manner. The antagonist methysergide blocked the effects of serotonin and quipazine on Prl. Acetylcholine and its agonist pilocarpine also stimulated release of Prl and inhibited release of GH in a concentration-related manner. Atropine blocked these responses. The results show that serotonin and acetylcholine affect pituitary hormone secretion by acting on the hypothalamus. They may stimulate the secretion of a Prl releasing hormone and somatostatin.

The deprivation of water for 12 or 24 hr increased the prolactin concentration in the plasma of immature chickens but had no effect on the circulating growth hormone (GH) level. The increase in plasma prolactin level reflected an increase in the basal rate of prolactin release from incubated hemipituitary glands and an increase in the responsiveness of the pituitary gland to hypothalamic releasing factors. The deprivation of water had no effect on basal level of pituitary GH release in vitro but abolished the stimulatory effect of the hypothalamus on in vitro GH secretion.

Chicken pituitary glands were incubated in medium containing oestradiol 17 beta (E2), alone or together with single whole hypothalami. E2 stimulated prolactin release from the pituitary and increased the prolactin releasing activity of the hypothalamus, but did not affect growth hormone release. Preincubation of pituitaries with E2 dramatically stimulated subsequent prolactin release. Pituitaries primed with E2 were more responsive to the prolactin-stimulating effects of hypothalamic extract (HE) and thyrotrophin-releasing hormone (TRH) and more sensitive to the prolactin-inhibiting effect of dopamine. E2-primed pituitaries were much less sensitive to the growth hormone releasing activity of TRH and HE. These results show that E2 may regulate pituitary function by direct effects on hormone release by modifying pituitary sensitivity to stimulatory or inhibitory influences and by altering hypothalamic releasing activity.

Anterior pituitary glands from chickens (Gallus domesticus) were incubated with or without single, mediobasal chicken hypothalami in medium containing histamine, alone or together with the antagonist diphenhydramine or in medium containing gamma-aminobutyric acid (GABA), alone or together with the antagonists bicuculline or picrotoxin. The release of prolactin (Prl) and growth hormone (GH) was measured by homologous radioimmunoassay. Histamine had no direct effect on the release of either hormone but stimulated Prl (in a dose-related way) and GH release when anterior pituitary glands were co-incubated with hypothalami. Diphenhydramine also had no direct effect on Prl or GH secretion but blocked the stimulatory effect of histamine on hypothalamus-induced Prl and GH release. When anterior pituitary glands were incubated without hypothalami, GABA, bicuculline and picrotoxin had no effect on the release of Prl or GH. However, GABA inhibited the release of both hormones in a concentration-related manner, when anterior pituitary glands were co-incubated with hypothalami. This inhibition was blocked by both bicuculline and picrotoxin. These results suggest that histamine and GABA may be involved in controlling the secretion of Prl and GH from the avian pituitary gland, possible by modifying the secretion of hypothalamic releasing and/or release-inhibiting hormones.

Pituitary glands and hypothalami from broiler fowl were incubated in medium containing testosterone, and prolactin and GH release were determined. Pituitary glands were also preincubated for 20 h in medium containing testosterone, and then in medium containing various secretagogues. Testosterone inhibited the release of prolactin directly from the pituitary gland in a concentration-related manner. The hypothalamus stimulated the release of prolactin, but by a lesser amount in the presence of testosterone. When pituitary glands were preincubated with testosterone, subsequent release of prolactin was inhibited, except with the highest concentration which stimulated prolactin release. Hypothalamic extract (HE) markedly stimulated prolactin release from control pituitary glands although testosterone-primed glands were less responsive. The stimulation of prolactin release by thyrotrophin releasing hormone (TRH) and prostaglandin E2 (PGE2) was also reduced by preincubation of the pituitary glands with testosterone. Priming with testosterone did not affect the release of GH from pituitary glands alone, but reduced the TRH-, HE- and PGE2-stimulated release of GH. These results demonstrate that testosterone directly inhibits prolactin secretion and reduces the sensitivity of pituitary lactotrophs and somatotrophs to provocative stimuli.

Cockerels were injected with drugs which affect brain serotoninergic activity. Concentrations of plasma prolactin and growth hormone have been measured and correlated with hypothalamic serotonin and 5-hydroxyindole acetic acid (5HIAA) levels. Tryptophan concentrations in the brain and the activity of monoamine oxidase (MAO types A and B) were also measured in some studies. The administration of the MAO inhibitor, pargyline, produced dose- and time-related reductions in brain MAO type A and B activities, hypothalamic 5HIAA concentrations and plasma growth hormone levels, but increased the hypothalamic serotonin and plasma prolactin concentrations. Clorgyline administration inhibited MAO type A (but not type B) activity and also increased hypothalamic serotonin and plasma prolactin levels, while reducing hypothalamic 5HIAA and circulating growth hormone concentrations. Deprenyl treatment inhibited MAO type B (but not type A) activity but did not significantly affect serotonin, 5HIAA, prolactin or growth hormone levels. The serotonin precursor, tryptophan, elevated brain tryptophan levels when given systemically. It also increased hypothalamic serotonin in a dose-related manner, increased plasma prolactin and reduced plasma growth hormone concentrations. Pargyline and tryptophan treatments affected hormone levels more markedly in 3-week-old than in 18-week-old cockerels. These results demonstrate a strong relationship between hypothalamic serotoninergic activity, MAO type A activity and the secretion of prolactin and growth hormone in the cockerel.