Expression of D1 and D2 dopamine receptors in the hypothalamus and pituitary during the turkey reproductive cycle: colocalization with vasoactive intestinal peptide

The regulation of avian prolactin (PRL) secretion and PRL gene expression is influenced by hypothalamic vasoactive intestinal peptide (VIP), the PRL-releasing factor in avian species. Recent evidence indicates that D(1) and D(2) dopamine (DA) receptors play a pivotal role in VIP and PRL secretion. The differential expression of DA receptors located on hypothalamic VIP neurons and anterior pituitary cells may affect the degree of prolactinemia observed during the turkey reproductive cycle. The relative expression of D(1D) and D(2) DA receptor subtype mRNA was quantitated using in situ hybridization histochemistry (ISH). D(1D) and D(2) DA receptor mRNA was found expressed throughout the hypothalamus and pituitary. The expression of D(1D) DA receptor mRNA in the hypothalamus was found to be 6.8-fold greater than that of D(2) DA receptor mRNA. Higher D(1D) DA receptor mRNA content was found in the anterior hypothalamus (3.6-fold), the ventromedial nucleus (2.0-fold), the infundibular nuclear complex (INF; 1.9-fold), and the medial preoptic nucleus (1.5-fold) of laying hens as compared to that of reproductively quiescent non-photostimulated hens. The levels seen in incubating hyperprolactinemic hens were essentially the same as in laying hens, except for the INF where levels were 52% higher. During the photorefractory stage (hypoprolactinemia), the D(1D) DA receptor mRNA was at its lowest level in all areas tested. No differences were observed in hypothalamic D(2) DA receptor mRNA abundance throughout the reproductive cycle, except for an increase in D(2) DA receptor mRNA within the INF of photorefractory hens. Also, a marked reduction in D(2) DA receptor mRNA was observed in the pituitary of incubating hens. Pituitary D(1D) DA receptor levels did not change when birds entered the incubating phase. Double ISH revealed that D(1D) and D(2) DA receptor mRNAs were co-expressed within neurons expressing VIP mRNA, predominantly within the lateral hypothalamus and INF. D(1D) DA receptor mRNA was more highly expressed than D(2) DA receptor mRNA. The present findings clearly demonstrate that the expression of stimulatory D(1) DA receptor mRNA in the hypothalamus increases in hyperprolactinemic incubating hens, whereas inhibitory D(2) DA receptor mRNA increases in the pituitary of hypoprolactinemic photorefractory hens.     

Effects of reproductive status, ovariectomy, and photoperiod on vasoactive intestinal peptide in the female turkey hypothalamus.

Vasoactive intestinal peptide (VIP) appears to be a physiologically relevant prolactin (PRL)-releasing factor during the avian reproductive cycle, yet little is known of the factors involved in modulating the hypothalamic concentrations of this neuropeptide. A heterologous chicken VIP radioimmunoassay was developed to examine the effects of reproductive status, ovariectomy, and photoperiod on hypothalamic VIP immunoreactivity in the female turkey. VIP concentrations were highest in the median eminence/infundibular nuclear complex (ME/INF) relative to other subregions of the hypothalamus and changed only in this region during the reproductive cycle. Quiescent, nonphotostimulated hens subjected to stimulatory photoperiod exhibited a 1.6-fold increase in VIP in the ME/INF (quiescent 59.9 +/- 6.0 vs photostimulated 95.8 +/- 7.1 pg/microgram protein). An additional 2-fold increase in ME/INF VIP concentrations was observed in laying hens (183.0 +/- 28.5 pg/microgram protein). Coincident increases in plasma PRL were also observed. In contrast, during incubation and the photorefractory stage, a dissociation between hypothalamic VIP and plasma PRL occurred. No changes were observed in VIP in incubating hens, yet a 6-fold increase in PRL was noted, compared to layers. In addition, ME/INF VIP concentrations exhibited no change during the photorefractory stage, whereas a 28-fold decrease in plasma PRL occurred. VIP concentrations in the ME/INF of laying hens were unaffected by ovariectomy, whereas exposure to short photoperiod reduced VIP by 44%. The inhibitory effects of short photoperiod could not be reversed by administration of exogenous steroids, while steroid treatment reduced VIP concentrations by 45% in the ovariectomized hens. These results provide additional correlative evidence for a modulatory role of VIP in PRL secretion and suggest that the expression of this neuropeptide in the INF may serve as a neural link between photoperiodic mechanisms and PRL release during the avian reproductive cycle.     

Alterations in hypothalamic vasoactive intestinal peptide-like immunoreactivity are associated with reproduction and prolactin release in the female turkey

Vasoactive intestinal peptide (VIP) has potent PRL-releasing activity, but its physiological role in the regulation of PRL release during the avian reproductive cycle is not known. We used indirect immunofluorescence to determine if changes in hypothalamic VIP are associated with the shifts in circulating PRL during the reproductive cycle of the domestic turkey. In the naturally hyperprolactinemic incubating hen, the majority of VIP immunoreactivity (VIP-IR) existed within neurons of the infundibular nuclear complex (INF) and fibers in the external layer of the median eminence. Within the INF, the numbers of VIP-IR cells increased during the cycle, paralleling increases in serum PRL. In the reproductively inactive, nonphotostimulated hen with low serum PRL, essentially no positive cells were noted, whereas the incubating hen exhibited 32.1 +/- 2.2 cells/pair of adjacent sections in the anterior INF and 59.6 +/- 2.0 cells in the posterior INF. Exposure of inactive hens to a stimulatory photoperiod resulted in a 2.6-fold increase in serum PRL with the appearance of VIP-IR cells in the INF. During laying and incubation, further increases were observed in the number of positive cells in the INF and serum PRL as well as a greater fiber density in the median eminence. To further examine the association between changes in VIP-IR and serum PRL, circulating PRL was artificially lowered by depriving incubating hens of their nests for 0, 2, 5, and 10 days. On day 2 of nest deprivation, serum PRL declined markedly to 12% of day 0 levels, with VIP-IR cell numbers at 64% and 46% in the anterior and posterior INF, respectively. By day 10, birds exhibited cell numbers in the INF averaging 20% of those observed in the day 0 incubating hens, with serum PRL at 6% of day 0 levels. The results of these studies indicate a possible causal relationship between hypothalamic VIP and changes in PRL secretion during the avian reproductive cycle, providing a basis for further research on the importance of this peptide as well as factors responsible for the modulation of its expression in hypothalamic INF neurons.     

Dopamine infusion into the third ventricle increases gene expression of hypothalamic vasoactive intestinal peptide and pituitary prolactin and luteinizing hormone beta subunit in the turkey

Turkey prolactin (PRL) secretion is controlled by vasoactive intestinal peptide (VIP) neurons residing in the infundibular nuclear complex (INF) of the hypothalamus. The VIPergic activity is modulated by dopamine (DA) via stimulatory D(1) DA receptors. DA (10 nmol/min for 40 min) was infused into the third ventricle of laying turkey hens to study its effect on circulating PRL, hypothalamic VIP and pituitary PRL and LHbeta subunit mRNA levels. Plasma PRL was significantly elevated after 20 min of DA infusion and remained elevated 30 min after cessation of infusion. Hypothalamic VIP mRNA content was significantly greater in the INF of DA-infused birds than it was in the INF of vehicle-infused control birds. No increase in VIP mRNA due to DA infusion was noted in the preoptic area. Pituitary PRL and LHbeta subunit mRNAs were increased in DA-infused hens as compared to vehicle-infused controls but the rate of increase was more in PRL than LHbeta subunit. This study demonstrates that exogenous DA activates hypothalamic VIP gene expression and this increased expression is limited exclusively to the avian INF. The increased VIP mRNA in the INF is correlated with increased levels of circulating PRL and PRL and LHbeta mRNAs in the anterior pituitary.     

Maternal adrenalectomy abrogates the effect of fetal alcohol exposure on the interleukin-1beta-induced febrile response: gender differences

Avian prolactin (PRL) secretion is regulated by vasoactive intestinal peptide (VIP) neurons residing in the infundibular nuclear complex (INF) of the hypothalamus. This VIPergic activity is modulated by stimulatory dopaminergic inputs. Dynorphin, serotonin (5-HT), dopamine (DA) and VIP all appear to stimulate PRL secretion along a hypothalamic pathway, expressing kappa opioid, serotonergic, dopaminergic and VIPergic receptors in succession, with the VIPergic system as the final mediator. Electrical stimulation (ES) within the turkey hypothalamus at the level of the medial preoptic area (POA), the ventromedial hypothalamic nucleus (VMN), the INF or the median eminence (ME) results in the release of PRL. When the selective D(1) DA receptor antagonist SCH-23390 HCl was infused intraventricularly at the rate of 10 nmol/min, ES in the POA or VMN was unable to increase PRL levels, while ES in the INF and ME did increase PRL to the same level as that of controls. These results were interpreted to suggest that the D(1) DA receptors involved in PRL release lie caudally to the VMN and dorsally to the INF. Bilateral microinjections (50 ng) of the D(1) DA receptor agonist SKF-38393 HCl into the POA or VMN failed to produce any increase in PRL, while similar microinjections in the INF increased PRL significantly within 15 min. Bilateral microinjections of the D(1) DA antagonist (50 ng) into the INF blocked the rise in PRL associated with ES in the POA. Bilateral microinjections of a D(2) DA antagonist (50 ng) into the INF failed to block PRL secretion induced by ES in the POA. Tract tracing, using double-label immunocytochemistry, revealed the presence of a monosynaptic dopaminergic pathway projecting from the POA to the INF. These data imply that the only hypothalamic D(1) DA receptors involved in the regulation of avian PRL secretion are those residing within the INF in the same region as the VIP neurons known to be involved in PRL secretion.     

Vasoactive Intestinal Peptide Concentrations in Turkey Hypophysial Portal Blood Differ across the Reproductive Cycle

Hypophysial portal blood was collected for the first time in an avian species using a dorsal approach through the third ventricle and median eminence of the turkey. This was done to test for the presence of the prolactin (PRL)-releasing factor vasoactive intestinal peptide (VIP) in the portal blood and to determine whether VIP concentrations there varied with and corresponded to plasma PRL levels across the reproductive cycle. VIP concentrations in hypophysial portal blood were 2.5- to 13.5-fold greater than in the general circulation. VIP concentrations were lowest in portal blood of nonphotostimulated, reproductively inactive hens (231.8 ± 26.4 pg/ml) and highest in incubating hens (1108.1 ± 363.7 pg/ml), while laying, and photorefractory hens were intermediate at 372.5 ± 95.6 and 715.3 ± 338.5 pg/ml, respectively. These differences in concentration of VIP in portal blood mirrored those of PRL in the general circulation and support other evidence that VIP is the avian PRL-releasing factor.     

The relative importance of vasoactive intestinal peptide and peptide histidine isoleucine as physiological regulators of prolactin in the domestic turkey

In mammals, prolactin (PRL) secretion is regulated by vasoactive intestinal peptide (VIP) and peptide histidine isoleucine (PHI). In birds, however, VIP is considered a PRL-releasing factor (PRF), while the role of PHI is unknown. The purpose of this study was to compare the effects of turkey PHI (tPHI) and turkey VIP (tVIP) on PRL secretion in vitro, and to study their physiological significance in vivo through active immunization against tPHI and tVIP. In vitro studies were conducted using pituitary cell cultures from female turkeys. In the in vivo study, female turkeys were immunized with keyhole limpet hemocyanin (KLH; control), synthetic tPHI conjugate (KLH-tPHI), or synthetic tVIP conjugate (KLH-tVIP). Both tVIP and tPHI stimulated PRL secretion from anterior pituitary cells in a dose response manner. However, tPHI was 100-fold less potent than tVIP in stimulating maximum PRL secretion in vitro. In addition, the highest dose (10(-4) M) of tPHI inhibited its own PRL-releasing activity as well as that of VIP-stimulated PRL release. Whereas, circulating PRL levels and nesting activity remained low and unchanged during the photo-induced reproductive cycle (i.e., experimental period) in tVIP-immunized birds, control and tPHI-immunized turkeys showed a significant increase in plasma PRL levels and in the incidence of incubation behavior over time following photostimulation. These findings, taken together with earlier results, indicate that VIP is the sole physiological PRF in the turkey (avian species).     

Enhanced vasoactive intestinal peptide-induced prolactin secretion from anterior pituitary cells of incubating turkeys (Meleagris gallopavo)

During incubation, female turkeys exhibit elevated circulating prolactin (PRL) which may be the result of enhanced pituitary responsiveness to vasoactive intestinal peptide (VIP). This hypothesis was tested by comparison of spontaneous and porcine VIP-induced PRL secretion from anterior pituitary cells of hens in various reproductive conditions. The effect of VIP and luteinizing hormone releasing hormone (LHRH), alone and in combination, on luteinizing hormone (LH) secretion was also examined. Incubation with pVIP (10(-10) to 10(-6) M) significantly stimulated PRL secretion at all incubation times tested (1-5 hr). This increase was greatest in cells from incubating hens, with those from laying, photorefractory, and quiescent (nonphotostimulated) hens secreting successively less PRL. These responses were obtained when spontaneous PRL secretions were compared. VIP induced approximately a similar 1.5-fold increase in LH secretion, in all reproductive groups. Also, VIP enhanced LHRH-induced LH secretion (1.2- to 1.6-fold; P less than 0.0001). It is concluded that PRL secretion in vitro by pituitary cells from turkey hens in various reproductive stages reflects the circulating levels of PRL at these stages.     

Expression of hypothalamic GnRH-I mRNA in the female turkey at different reproductive states and following photostimulation

In birds, changes in hypothalamic gonadotropin-releasing hormone-I (GnRH-I) content and release are correlated with reproductive stages. This study examined the distribution and expression level of GnRH-I mRNA in anatomically discrete hypothalamic nuclei throughout the turkey reproductive cycle and following photostimulation. GnRH-I mRNA expression was determined using in situ hybridization in non-photostimulated (NPS), egg-laying (LAY), incubating (INC) and photorefractory (REF) hens. Overall, GnRH-I mRNA expression was greatest in the nucleus commissurae pallii (nCPa) and around the organum vasculosum lamina terminalis (OVLT), with less expression observed in the nucleus septalis lateralis (SL), cortico-habenula cortico-septum area, and within the nucleus preopticus medialis. GnRH-I mRNA expression was significantly increased in nCPa, OVLT, and SL after NPS hens (6L:18D) were exposed to a 30 or 90 min pulse of light beginning 14 h after first light (dawn). GnRH-I mRNA abundance within nCPa, OVLT and SL was greater in LAY than in NPS and INC hens, while mRNA expression was least in REF hens. These results indicate that GnRH-I mRNA expression in birds is sensitive to light stimulation during the photosensitive period and can be used to more precisely characterize their different reproductive stages.     

Evidence of a role for the turkey posterior pituitary in prolactin release.

The aims of this study were: (1) to examine whether the posterior pituitary contains prolactin releasing factor (PRF) activity, (2) to determine to what extent known neurohypophyseal peptides contribute to this activity, and (3) to compare posterior pituitary PRF activities of hens in different reproductive stages. Anterior pituitary cells derived from juvenile female turkeys were incubated with posterior pituitary extracts or test substances for 3 hr. Posterior pituitary extracts (0.1-0.8 equivalent) contained a potent substance(s) which stimulated PRL release in a concentration-dependent manner (2.4 +/- 0.08 to 6.5 +/- 0.23 micrograms/500 k cells). Arginine vasotocin (AVT) and vasoactive intestinal peptide (VIP) antisera (1:500) completely abolished the PRL-releasing activities of their respective peptides but partially reduced (P less than 0.05) the PRF activity of the posterior pituitary (AVT, 19.9%; VIP, 55.1%). Mesotocin antiserum did not alter (P greater than 0.05) PRL release induced by posterior pituitary extract. Posterior pituitary extract (0.01-0.5 equivalent) from hens in each of the various stages of the reproductive cycle induced a concentration dependent PRL release. The 0.5 posterior pituitary equivalent dose from reproductively quiescent (nonphotostimulated), laying, photorefractory, and incubating hens increased PRL release 2.4-, 2.9-, 3.8-, and 11.1-fold, respectively. The turkey posterior pituitary contains a potent PRF activity, partially accounted for by VIP and AVT, at the assayed concentrations, which varies with the reproductive cycle.     

Involvement of dopamine in prolactin release induced by electrical stimulation of the hypothalamus of the female turkey (Meleagris gallopavo).

Controversy exists regarding the role of dopamine (DA) in the regulation of avian prolactin (PRL) secretion. Consequently, we injected apomorphine, a DA agonist, and pimozide, a DA receptor blocker, into laying and nest-deprived incubating turkeys and studied their effect on PRL secretion before (-20, -10, 0 min), during (5, 10, 20, 30 min), and after (5, 15, 30 min) electrical stimulation in the ventromedial nucleus of the hypothalamus. Apomorphine (10 mg/kg, ip) completely abolished the electrical stimulation-induced PRL increase in both laying and nest-deprived incubating hens. Pimozide (2 mg/kg, ip) potentiated electrical stimulation-induced PRL secretion in laying hens. In the two pimozide experiments, peak responses were 10.9-fold for the pimozide-treated group vs 2.9-fold for the control group, and 5.4-fold for the pimozide-treated group vs 2.6-fold for the control group. In nest-deprived incubating hens, PRL response to electrical stimulation was unaffected by pimozide treatment. These data support the concept that DA is inhibitory to the neuroendocrine system which stimulates PRL secretion in laying hens. In incubating hens, the dopaminergic inhibition is diminished, allowing for the increased PRL level observed during incubation.     

Cloning of a turkey prolactin cDNA: expression of prolactin mRNA throughout the reproductive cycle of the domestic turkey (Meleagris gallopavo)

A cDNA-encoding turkey prolactin (PRL) has been isolated from a turkey pituitary library. The 953-base pair cDNA clone contains a 229-amino acid open reading frame which consists of a 30-amino acid signal peptide followed by a 199-amino acid mature PRL. The deduced amino acid sequence of turkey PRL shows greater than 90% homology to chicken PRL and 54-78% homology to other mammalian prolactins. A mRNA of 1100 nucleotides was detected in total RNA extracted from turkey pituitaries. Levels of PRL mRNA increased approximately 10-, 20-, and 100-fold in photostimulated, laying, and incubating hens, respectively, relative to that found in nonphotostimulated hens. The corresponding increases in plasma PRL levels were 2-, 5.5-, and 50-fold and in pituitary PRL content were 2-, 4-, and 13.4-fold, respectively. The transition from incubation to the photorefractory phase resulted in a 10-fold reduction in PRL mRNA, a 3.7-fold decrease in pituitary PRL, and a dramatic 50-fold decrease in plasma PRL. The changes in the abundance of pituitary PRL mRNA appear to be related to the changes in PRL-releasing activity observed at each of the reproductive stages. This study provides the first characterization of pituitary PRL mRNA and its comparison with plasma and pituitary PRL levels during the avian reproductive cycle.     

Prolactin release and response to vasoactive intestinal peptide in an opportunistic breeder, the zebra finch (Taeniopygia guttata)

Zebra finches in arid regions of Australia are opportunistic breeders that time their breeding cycles to coincide with nonseasonal rainfall. Hormonal profiles associated with reproductive behaviors may differ from those observed in seasonal breeders because these birds need to be reproductively competent on short notice. This study measured plasma prolactin (PRL) levels in nonbreeding and breeding zebra finches and in birds with and without prior reproductive experience. We also investigated the change in plasma PRL following injection with vasoactive intestinal peptide (VIP), the avian PRL-releasing hormone. PRL was lowest in non-paired birds, increased after pair bonds had formed, and was highest in incubating birds. No differences in PRL levels were found between males and females in these biparental care-givers. A single injection of VIP resulted in a rapid increase in plasma PRL in nonbreeding zebra finches, while PRL remained unchanged in incubating birds. When escalating doses of VIP were administered, nonbreeders responded with a maximal response in PRL release, but PRL levels in breeders remained unchanged following even the highest VIP dose. Among nonbreeders, inexperienced birds had significantly lower PRL levels than birds that had successfully reared a clutch, but both groups responded with an equally robust increase in PRL following a VIP challenge. This pattern differs from that observed in most photosensitive species in which only during a breeding cycle do birds secrete significant levels of PRL in response to exogenous VIP. Zebra finches, even when not actively breeding, must maintain competent pituitary lactotrophs that can secrete PRL at maximal rates. This is part of the suite of characters enabling these birds to respond to favorable breeding conditions at any time.     

Vasoactive intestinal peptide and peptide with N-terminal histidine and C-terminal isoleucine increase prolactin secretion in cultured rat pituitary cells (GH4C1) via a cAMP-dependent mechanism which involves transient elevation of intracellular Ca2+

Vasoactive intestinal peptide (VIP) and peptide (P) with N-terminal histidine and C-terminal isoleucine (PHI) stimulated prolactin (PRL) secretion from GH4C1 cells equipotent with ED50 values of 30-50 nM. In a parafusion system optimized to give high time resolution both VIP and PHI increased PRL secretion with a delay of about 60 s and subsequent to the activation of the adenylate cyclase. Thyroliberin (TRH) increased PRL secretion within 4 s. The dose-response curves for VIP- and PHI-stimulated cAMP accumulation were superimposable on those for PRL secretion. At submaximal concentrations the effects of VIP and PHI on both cAMP accumulation and PRL secretion were additive, whereas the effects were not additive at concentrations giving maximal effects. VIP and PHI increased [Ca2+]i measured by quin-2 in a different way than TRH, without inducing changes in the electrophysiological membrane properties of the GH4C1 cells. We conclude that both VIP and PHI stimulate PRL secretion via a cAMP-dependent process involving an increase in [Ca2+]i.     

The role of hypothalamic vasoactive intestinal polypeptide in the maintenance of prolactin secretion in incubating bantam hens: observations using passive immunization, radioimmunoassay and immunohistochemistry

The role of chicken vasoactive intestinal polypeptide (cVIP) as a prolactin-releasing factor was investigated in incubating bantam hens. Specific antibodies were raised against cVIP (anti-cVIP) for passive immunization studies, to develop a radioimmunoassay and to localize VIP neurones immunohistochemically in the hypothalamus. The concentration of plasma prolactin decreased after i.v. injection of anti-cVIP: this low concentration being maintained by daily injection of anti-cVIP. Incubating hens injected daily with anti-cVIP deserted their nests after 4.5 +/- 0.6 days and returned to lay after 20 +/- 1 days. This disruption of incubation behaviour with anti-cVIP was prevented by concomitant, twice daily, injections of 30 IU ovine prolactin. The concentration of plasma LH was not immediately affected after injection of anti-cVIP but increased when the hens deserted their nests. The amount of cVIP, measured by radioimmunoassay, was significantly higher in the median eminence (P less than 0.01) and medial basal hypothalamus (P = 0.05) in incubating than in laying hens. No differences were seen in the amounts of cVIP in the preoptic hypothalamus or in a part of the forebrain including the nucleus accumbens, between laying and incubating hens. Morphological observations were made on immunohistochemically identified cVIP cell bodies in the medial basal hypothalamus. These showed that cVIP cell number, cell area and density of immunoreactive product were significantly (P less than 0.05) greater in incubating than in laying hens. Further, the density of cVIP reaction product in the anterior median eminence was also significantly (P less than 0.01) greater in incubating than in laying hens.(ABSTRACT TRUNCATED AT 250 WORDS)     

Vasoactive intestinal peptide increases prolactin messenger ribonucleic acid content in GH3 cells

The influence of vasoactive intestinal peptide (VIP), TRH, and dexamethasone (Dex) on PRL mRNA was investigated in PRL-producing GH3 cells using cytoplasmic dot hybridization and RNA blot analysis. Total cytoplasmic RNA was transferred to nitrocellulose filter paper and quantitated by hybridization to PRL recombinant DNA probes labeled with 32P. Incubation of GH3 cells with VIP for 25 h increased the content of cytoplasmic PRL mRNA. This increase was dose dependent, being significant at 2 X 10(-8) M and reaching a maximum at 2 X 10(-7) M. VIP at 2 X 10(-9) M had no effect on cytoplasmic PRL mRNA content. TRH (2 X 10(-7) M) also increased whereas Dex (2 X 10(-7) M) decreased the content of PRL mRNA. The inhibitory effect of Dex (2 X 10(-7) M) on cytoplasmic PRL mRNA was reversed by VIP (2 X 10(-7) M). Changes in medium PRL levels after these various hormone treatments paralleled those changes observed in PRL mRNA content. Examination of total poly(A)+ RNA demonstrated that incubation with VIP (2 X 10(-7) M) for 6 h increased the content of the mature PRL mRNA and its processing intermediates. Dex (2 X 10(-7) M) decreased the content of all species of PRL mRNA. These data suggest that VIP-stimulated PRL release is the result of an increase in the content of PRL mRNA and its precursors.     

Changes in vasoactive intestinal peptide and tyrosine hydroxylase immunoreactivity in the brain of nest-deprived native Thai hen

Hyperprolactinemia is associated with incubation behavior and ovarian regression in birds. To investigate the association of prolactin (PRL), vasoactive intestinal peptide (VIP), and dopamine (DA) with the neuroendocrine regulation of incubation behavior, changes in the number of visible VIP-immunoreactive (VIP-ir) neurons in the nucleus inferioris hypothalami (IH) and nucleus infundibuli hypothalami (IN) and tyrosine hydroxylase-immunoreactive (TH-ir) neurons in the nucleus intramedialis (nI) and nucleus mamillaris lateralis (ML) of incubating native Thai hens were compared with those of nest-deprived hens. TH was used as a marker for dopaminergic (DAergic) neurons. Blood samples were collected to determine PRL levels. The localization and the number of visible VIP-ir and TH-ir neurons were determined by immunohistochemistry. Disruption of incubation behavior was accompanied by a precipitous decline in plasma PRL levels. The number of visible VIP-ir neurons in the IH-IN and TH-ir neurons in the nI and ML were high during incubation and decreased when hens were deprived of their nests. This study indicated an association between VIP neurons in the IH-IN and DA neurons in the nI and ML with the degree of hyperprolactinemia, suggesting that the expression of incubation behavior in birds might be, in part, regulated by the DAergic input from the nI and ML to VIP neurons in the IH-IN and subsequent PRL release.     

Involvement of serotonin in prolactin release induced by electrical stimulation of the hypothalamus of the turkey (Meleagris gallopavo)

In anesthetized female turkeys electrical stimulation of the ventromedial nucleus (VMN) for 30 min caused increases in plasma prolactin (Prl); with maximum increase above the prestimulation level being 33.6 +/- 5.7 ng/ml for laying hens and 768.1 +/- 187.6 ng/ml for incubating hens. The possibility that serotonin (5-HT) plays a role in electrical stimulation-induced Prl release was investigated after administration of methysergide, a 5-HT receptor blocker (20 mg/kg), and stimulation in either the VMN or the infundibular nuclear complex-median eminence (INF-ME) region. Electrical stimulation in both the VMN and INF-ME region caused increases (P less than 0.05) in plasma Prl. Pretreatment with methysergide prevented the increase in plasma Prl that follows electrical stimulation in the VMN but had no effect on electrical stimulation-induced Prl release in the INF-ME region. We conclude that Prl release in the female turkey requires the functional integrity of serotonergic neurons within the VMN.