J. A. Proudman

Stimulation of prolactin release in turkeys by vasoactive intestinal peptide.

Abstract Vasoactive intestinal peptide (VIP) is a potent releasor of prolactin in birds. The main purpose of this study was to identify its site of action. Synthetic porcine VIP administered intraatrially to freely moving ovariectomized (OVX) turkeys induced an elevation of circulating PRL within 15 min in a dose-related manner. Removal of hypothalamic control of PRL release by surgical disconnection of the neurohemal regions of the median eminence did not significantly diminish the PRL response to VIP. Intraatrial injection of eledoisin or bradykinin into OVX hens did not influence PRL secretion, indicating that the PRL releasing activity of VIP is probably not attributable to its vasodilatory action. These results support the possibility that VIP is an authentic prolactin releasing factor (PRF) in birds.

The turkey prolactin-encoding gene and its regulatory region ☆

Overlapping prolactin (Prl) lambda clones were isolated from a turkey genomic library. The 6.7-kb turkey Prl gene consists of five exons. Major transcription start points were located by primer extension 51-53 nucleotides upstream from the Met start codon. No estrogen response element (ERE) was found, but two regions similar to mammalian Pit-1/GHF-1-binding sites were identified by computer analysis. This suggests that transcription of the turkey Prl gene may be regulated by Pit-1/GHF-1, and not by the estrogen receptor.

Effects of calorie restriction on reproductive and adrenal systems in Japanese quail: Are responses similar to mammals, particularly primates?

The benefits of calorie restriction (CR) have been established across a variety of vertebrate and invertebrate species. Although the effects of CR on lifespan in birds have not been examined, it is clear that CR has beneficial effects on reproductive, metabolic, and physiological function in adult poultry. We examined the effects of CR in Japanese quail, a rapidly maturing avian model, on reproductive endocrine and neuroendocrine systems. Male Japanese quail were pair fed at 0% ad libitum (AL), 20%, or 40% CR of AL, recorded for juveniles (3-7 weeks of age) or adults (12-16 weeks of age). Juvenile males on CR matured more slowly, and both juvenile and adult males had reduced plasma luteinizing hormone (LH) with CR. Adults on 40% CR showed evidence of stress, with increased plasma corticosterone and reduced testes weight and circulating androgens. In a separate study, pituitary gland response was tested in juvenile and adult castrated males that had been on the same CR treatments. All birds responded to gonadotropin releasing hormone (GnRH) challenge, with LH release. However, the 40% CR juvenile and adult birds had quantitatively lower responses, suggesting central inhibition of the reproductive axis. This hypothesis was tested by measurement of sexual behavior and catecholamines known to stimulate GnRH in hypothalamic regions that modulate these responses. Results showed reduced norepinephrine in key hypothalamic regions and reduced dopamine in posterior hypothalamus. These data support the hypothesis that CR affects reproductive function, with evidence for effects in the central nervous system. These data are discussed and compared to data collected in mammals, especially the rhesus monkey, on the effects of timing and degree of CR on reproductive and stress responses.

Stimulation of prolactin secretion from turkey anterior pituitary cells in culture.

Abstract Prolactin (PRL) secretion by monolayer cultures of turkey anterior pituitary cells was significantly increased (up to 44-fold) by vasoactive intestinal peptide (VIP), arginine vasotocin (AVT), and by an extract of turkey hypothalami (HE). Several other neuropeptides (including thyrotropin-releasing hormone) and neurotransmitters were ineffective in influencing PRL secretion at doses up to 10-6 M. The dynamic PRL response to HE and VIP was studied using supervised pituitary cells attached to microcarrier beads. HE, administered in 30-min pulses, resulted in a significant, dose-related increase in PRL secretion from a basal secretion rate of 2.32 ng/min/107 cells to a peak secretion rate of 127.13 ng/min/107 cells at the highest dose of HE tested (1 mg tissue-equivalent weight/ml). VIP significantly increased PRL secretion at all doses studied (from 10-10 to 10-6 M), with 10-8 M VIP producing a response similar to that observed with 1 mg/ml HE. A highly significant (P < 0.001) linear relationship was demonstrated between the log-dose of VIP administered and peak PRL secretion rate. These studies suggest that VIP, but not TRH, may be a physiological stimulus for PRL release in the turkey.

Identification of Dopamine, Gonadotrophin-Releasing Hormone-I, and Vasoactive Intestinal Peptide Neurones Activated by Electrical Stimulation to the Medial Preoptic Area of the Turkey Hypothalamus: A Potential Reproductive Neuroendocrine Circuit

The neural and neurochemical substrates regulating reproduction in birds remain vaguely defined. The findings that electrical stimulation in the medial preoptic area (ES/MPOA) or intracerebroventricular infusion of dopamine (DA) stimulated luteinising hormone (LH) and prolactin (PRL) release in female turkeys, led to the suggestion that ES/MPOA might help to clarify the DA circuitry regulating LH and PRL. We used c‐fos mRNA and tyrosine hydroxylase immunoreactivity as measured by double in situ hybridisation/immunocytochemistry (ISH/ICC) to determine which group/subgroup of DA neurones was activated following unilateral ES/MPOA. To establish that the reproductive neuroendocrine system was activated, double ISH/ICC was also conducted on c‐fos/gonadotrophin‐releasing hormone‐I (GnRH‐I) and c‐fos/vasoactive intestinal peptide (VIP). Changes in circulating LH and PRL were determined by radioimmunoassay. Unilateral ES/MPOA (100 µA, right side) of anaesthetised laying turkeys for 30 min increased circulating LH and PRL levels. It also induced c‐fos mRNA expression on the ipsilateral side by all GnRH‐I neurones within the septopreoptic region, implying that GnRH‐I neurones in this region share similar circuitry. VIP neurones within the nucleus infundibularis were the only VIP group to show c‐fos mRNA expression, suggesting their involvement in ES/MPOA induced PRL release. c‐fos mRNA expression was also observed in a subgroup of DA neurones in the nucleus mamillaris lateralis (ML). To our knowledge, the present study is the first to show that activation of DAergic cells in the ML is associated with the activation of GnRH‐I and VIP neurones and the release of LH and PRL. It is likely that ES/MPOA activated VIP/GnRH‐I neurones via activation of DA neurones in the ML, as this was the only DA subgroup that showed c‐fos mRNA expression.

Pattern of secretion of luteinizing hormone and testosterone in the sexually mature male turkey

Whether luteinizing hormone (LH) and testosterone (T) are secreted in pulsatile patterns was determined in sexually mature male turkeys. Turkeys were chronically cannulated and serially bled for three 8-hr periods covering the 24-hr day (14L:10D, n = 7, series B), or for two 12-hr periods covering the 24-hr day (14L:10D, n = 4, series C). Pulses of both LH and T occurred during both the light and dark portions of the 24-hr day. A portion of the secretory episodes of T, where the baseline level of LH was relatively low, was associated with prior peaks of LH secretion. Secretory episodes of T also occurred, where baseline levels of LH and T were both relatively high, without detection of prior peaks of LH. No differences were found between the photophase and scotophase portions of the photoperiod for either LH or T concentration. It is concluded that T is secreted in a pulsatile pattern in sexually mature male turkeys. However, LH is secreted in a pulsatile pattern only when baseline levels of both LH and T are relatively low. Neither LH nor T secretion is entrained by the photoperiod. Corticosterone was measured in hourly samples, but no changes in concentration occurred in association with the photoperiod.

Exposure duration to long day lengths associated with the expression of photorefractoriness in turkey breeder hens

In addition to inducing egg production, exposure to long days concomitantly activates processes that eventually result in photorefractoriness (PR) and cessation of egg production. Experiments were conducted to evaluate the duration of exposure to long days that result in these processes. In each of 3 experiments, we subjected Large White turkey breeder hens to long days (16 or 18 h per day) for differing lengths of time from initial photostimulation and then returned them to a photoperiod (12L:12D) that provided sufficient, but decreased, photoperiodic drive to support egg production but not induce PR. Photoresponsiveness was then evaluated by egg production after a return to a longer day length (20 h per day) late in the lay period and beyond the mean onset of PR typical for these turkey hens. Hens that have undergone any reduction in photoresponsiveness should not increase egg production in response to the increased photoperiod. From experiments 1 and 2, exposure to long days for as little as 1 d and as much as 9 wk from initial photostimulation did not result in an alteration in subsequent photoresponsiveness. This was based on an increased egg production response to a change in photoperiod from 12L:12D to 20L:4D after 20 wk of photostimulation that was similar to controls held continuously on 12L:12D and opposite to the response of controls held continuously on 18L:6D. It was clear that PR had been fully programmed by 20 wk of exposure to long days. Exposure to long days for 12 wk (experiment 3) resulted in a partial alteration of subsequent photoresponsiveness. It was concluded that programming of PR during late spring-summer season occurs after 9 wk of long day exposure, is not fully expressed by 12 wk of long days, and can be fully expressed by 20 wk of photostimulation.

Turkey and chicken prolactins stimulate the proliferation of rat Nb2 lymphoma cells.

Abstract The purpose of this investigation was to assess the actions of two avian prolactins (PRL) on rat Nb2 lymphoma cell proliferation. Various turkey and chicken PRL preparations stimulated the proliferation of rat Nb2 cells. The ability of avian PRL preparations to influence the behavior of rat Nb2 lymphoma cells provides a useful method for monitoring the PRL-like actions of native and recombinant avian PRL and suggests a potential role of PRL in modulating the avian immune system.

Daily rhythm of prolactin and corticosterone in unrestrained, incubating turkey hens

Plasma concentrations of prolactin and corticosterone were determined in hourly samples collected over a 25-hr period from unrestrained turkey hens exhibiting incubation (broody) behavior. Hens were maintained in cages on a 14 hr light: 10 hr dark photoperiod. Mean plasma prolactin concentration increased significantly late in the dark period to reach maximum daily levels of 496 ng/ml at the start of the photophase. However, a well-defined daily rhythm in prolactin secretion was not evident. Plasma corticosterone concentration showed a significant daily rhythm characterized by a major secretory peak (7.14 ng/ml) in the middle of the light period and a smaller, less well-defined peak (4.11 ng/ml) during the dark period. Both prolactin and corticosterone secretion ranged widely throughout the day in individual hens in a manner suggestive of pulsatile hormone secretion. This study demonstrates that the extremely high prolactin concentrations characteristic of the incubating turkey hen reflect large, dynamic changes in plasma prolactin concentrations occurring throughout the photoperiod, rather than a constantly high level of prolactin secretion.

Differential regulation of gene expression and release of FSH and prolactin by long day and sulfamethazine in chicks.

In several avian species long day exposure results in plasma elevation of gonadotropins and prolactin (PRL). We examined the early (12-72h) effects of photostimulation on mRNA transcripts and plasma levels of follicle stimulating hormone (FSH) and PRL in three-week old cockerels. In addition, the neuroendocrine influence of the compound, sulfamethazine (SMZ), known to enhance light-induced gonadal development in chicks, was studied when applied with or without long-day photostimulation. Both long day exposure and SMZ intake caused a rapid increase in FSHbeta mRNA transcripts at Zeitgeber time 48 (ZT48), while only SMZ stimulated secretion of the hormone into plasma during the course of the study. In contrast to SMZ treatment, photostimulation was more effective at stimulating PRL mRNA transcripts and secretion of PRL. Results demonstrate a differential role of long day exposure and SMZ intake on the regulation of FSH and PRL synthesis and secretion and suggest that some effects of SMZ on gonadal development may be mediated by the pituitary.