Anatomical and functional implications of corticotrophin-releasing hormone neurones in a septal nucleus of the avian brain: an emphasis on glial-neuronal interaction via V1a receptors in vitro.

Previously, we showed that corticotrophin-releasing hormone immunoreactive (CRH-IR) neurones in a septal structure are associated with stress and the hypothalamic-pituitary-adrenal axis in birds. In the present study, we focused upon CRH-IR neurones located within the septal structure called the nucleus of the hippocampal commissure (NHpC). Immunocytochemical and gene expression analyses were used to identify the anatomical and functional characteristics of cells within the NHpC. A comparative morphometry analysis showed that CRH-IR neurones in the NHpC were significantly larger than CRH-IR parvocellular neurones in the paraventricular nucleus of the hypothalamus (PVN) and lateral bed nucleus of the stria terminalis. Furthermore, these large neurones in the NHpC usually have more than two processes, showing characteristics of multipolar neurones. Utilisation of an organotypic slice culture method enabled testing of how CRH-IR neurones could be regulated within the NHpC. Similar to the PVN, CRH mRNA levels in the NHpC were increased following forskolin treatment. However, dexamethasone decreased forskolin-induced CRH gene expression only in the PVN and not in the NHpC, indicating differential inhibitory mechanisms in the PVN and the NHpC of the avian brain. Moreover, immunocytochemical evidence also showed that CRH-IR neurones reside in the NHpC along with the vasotocinergic system, comprising arginine vasotocin (AVT) nerve terminals and immunoreactive vasotocin V1a receptors (V1aR) in glia. Hence, we hypothesised that AVT acts as a neuromodulator within the NHpC to modulate activity of CRH neurones via glial V1aR. Gene expression analysis of cultured slices revealed that AVT treatment increased CRH mRNA levels, whereas a combination of AVT and a V1aR antagonist treatment decreased CRH mRNA expression. Furthermore, an attempt to identify an intercellular mechanism in glial-neuronal communication in the NHpC revealed that brain-derived neurotrophic factor (BDNF) and its receptor (TrkB) could be involved in the signalling mechanism. Immunocytochemical results further showed that both BDNF and TrkB receptors were found in glia of the NHpC. Interestingly, in cultured brain slices containing the NHpC, the use of a selective TrkB antagonist decreased the AVT-induced increase in CRH gene expression levels. The results from the present study collectively suggest that CRH neuronal activity is modulated by AVT via V1aR involving BDNF and TrkB glia in the NHpC.

Distribution of the Vasotocin Subtype Four Receptor (VT4R) in the Anterior Pituitary Gland of the Chicken, Gallus gallus, and its Possible Role in the Avian Stress Response.

The neurohormone arginine vasotocin (AVT) in non mammalian vertebrates is homologous to arginine vasopressin (AVP) in mammals. Its actions are mediated via G protein-coupled receptors that belong to the vasotocin/mesotocin family. Because of the known regulatory effects of nonapeptide hormones on anterior pituitary functions, receptor subtypes in that family have been proposed to be located in anterior pituitary cells. Recently, an avian vasotocin receptor subtype designated VT4R has been cloned, which shares 69% sequence homology with a human vasopressin receptor, the V1aR. In the present study, a polyclonal antibody to the VT4R was developed and validated to confirm its specificity to the VT4R. The antibody was used to test the hypothesis that the VT4R is present in the avian anterior pituitary and is specifically associated with certain cell types, where its expression is modulated by acute stress. Western blotting of membrane protein extracts from pituitary tissue, the use of HeLa cells transfected with the VT4R and peptide competition assays all confirmed the specificity of the antibody to the VT4R. Dual-labelling immunofluorescence microscopy was utilised to identify pituitary cell types that contained immunoreactive VT4R. The receptor was found to be widely distributed throughout the cephalic lobe but not in the caudal lobe of the anterior pituitary. Immunoreactive VT4R was associated with corticotrophs. Approximately 89% of immunolabelled corticotrophs were shown to contain the VT4R. The immunoreactive VT4R was not found in gonadotrophs, somatotrophs or lactotrophs. To determine a possible functional role of the VT4R and previously characterised VT2R, gene expression levels in the anterior pituitary were determined after acute immobilisation stress by quantitative reverse transcriptase-polymerase chain reaction. The results showed a significant increase in plasma corticosterone levels (three- to four-fold), a significant reduction of VT4R mRNA and an increase of VT2R mRNA (P < 0.05) in acutely immobilised chicks compared to controls. The data suggest a role of the VT4R in the avian stress response.

Appetitive and consummatory sexual and agonistic behaviour elicits FOS expression in aromatase and vasotocin neurones within the preoptic area and bed nucleus of the stria terminalis of male domestic chickens.

Some components of male sexual and agonistic behaviours are considered to be regulated by the same neurocircuitry in the medial preoptic nucleus (POM) and the medial portion of bed nucleus of the stria terminalis (BSTM). To better understand this neurocircuitry, numbers of aromatase- (ARO) or arginine vasotocin- (AVT) immunoreactive (ir) neurones expressing immediate early gene protein FOS were compared in the POM and BSTM of male chickens following sexual or agonistic behaviours. Observations were made on males showing: (i) appetitive (courtship) and consummatory (copulation) sexual behaviours; (ii) only appetitive sexual behaviour, or (iii) displaying agonistic behaviour toward other males. Control males were placed on their own in the observation pen, or only handled. In the POM, appetitive sexual behaviour increased ARO+FOS colocalisation, whereas agonistic behaviour decreased the number of visible ARO-ir cells. In the dorsolateral subdivision of BSTM (BSTM1), appetitive sexual behaviour also increased ARO+FOS colocalisation, although the numbers of visible ARO-ir and AVT-ir cells were not altered by sexual or agonistic behaviours. In the ventromedial BSTM (BSTM2), appetitive sexual behaviour increased ARO+FOS and AVT+FOS colocalisation, and all behaviours decreased the number of visible ARO-ir cells, particularly in males expressing consummatory sexual behaviour. Positive correlations were found between numbers of cells with ARO+FOS and AVT+FOS colocalisation in both subdivisions of the BSTM. Waltzing frequency was positively correlated with ARO+FOS colocalisation in the lateral POM, and in both subdivisions of the BSTM in males expressing sexual behaviour. Waltzing frequency in males expressing agonistic behaviour was negatively correlated with the total number of visible ARO-ir cells in the lateral POM and BSTM2. These observations suggest a key role for ARO and AVT neurones in BSTM2 in the expression of appetitive sexual behaviour, and differential roles for ARO cells in the POM and BSTM in the regulation of components of sexual and agonistic behaviours.

Molecular neuroendocrine events during stress in poultry.

Magnocellular neurons in the hypothalamic paraventricular nucleus containing arginine vasotocin (AVT) project to the posterior pituitary and release the peptide peripherally to target tissues. Parvocellular neurons contain either corticotropin-releasing hormone (CRH), AVT, or both neuropeptides and project to the median eminence. Corticotropin-releasing hormone and AVT are then transported to the anterior pituitary where they bind to CRH1 or vasotocin VT2 receptors, many found co-localized on the same pituitary cell type, the corticotrope. Central administration of CRH compared with AVT is more effective in releasing the stress hormone corticosterone, whereas peripheral administration of AVT is more efficacious. Simultaneous, peripheral administration of CRH and AVT also resulted in a synergistic release of corticosterone. Cell culture studies demonstrated a synergistic release of the second messenger, cyclic adenosine monophosphate, when both CRH and AVT were added to cells transfected with CRH and VT2 receptors, providing a possible explanation for the enhanced release of corticosterone following combined peripheral administration of the 2 peptides. A social stress model, mature male-male interaction, demonstrated activation of neurons in the paraventricular nucleus and suggested that the posterior pituitary as well as the anterior pituitary are involved in a social stress response.

A landmark contribution to poultry science--a possible mode of action of sulfamethazine on the reproductive system of Leghorn cockerels.

The paper "The effect of sulfamethazine feeding on the thyroids, combs and testes of Single Comb White Leghorn cockerels" was published early in the career of Ari van Tienhoven when he was an assistant professor. The current manuscript pays tribute to that study submitted to the journal, Poultry Science, over 5 decades ago. In addition, the review examines the academic lineage of van Tienhoven to show that his line of descent produced several scientists who have contributed significantly to the success of the Poultry Science Association in the discipline of neuroendocrinology and avian physiology during most of its first 100 yr. In a quiet, unassuming manner, van Tienhoven has had a major effect on basic research addressing reproductive endocrinology in vertebrates, particularly in the domestic fowl, Gallus gallus. Biographical information, his academic pedigree, students produced, research areas pursued, and the landmark contribution selected for review are highlighted. A link to an electronic archive of a recorded interview with van Tienhoven in preparation for the symposium can be accessed using the link given in the first footnote.

Comparison of beak-trimming methods on early broiler breeder performance.

Beak trimming is necessary in commercial broiler breeders to prevent or decrease trauma as they mature. Two common beak-trimming methods were evaluated by early performance comparison with nontrimmed chicks (NBT). The robotic electrocautery device (ECD) trims and cauterizes the beak tip. The robotic infrared beak-trimming device (IBT) applies an infrared light beam to destroy the live basal tissue while leaving the hard corneum intact for the first approximately 10 d. In 2 experiments, day-of-hatch Ross 708 by-product chicks were obtained from a local hatchery, where 1/3 of the chicks were trimmed using IBT. All chicks were then transported to another hatchery where 1/3 were trimmed using ECD and 1/3 were NBT. Personnel at each hatchery were highly experienced and skilled with their respective technique. All chicks were then transported to University of Arkansas facilities. Before placement in each experiment, chicks were individually neck-tagged and weighed, and in experiment 1, beaks were measured using a digital caliper. A small but significant transient reduction in BW gain was observed at 14 d due to ECD as compared with NBT controls, although ECD was not different than IBT in experiment 1. In experiment 2, IBT birds were significantly heavier at 11 d by 7.8 and 8.7 g than the NBT or ECD, respectively. However, at d 21 and 42, no significant differences in BW or BW gain were observed. When beak trimming was performed on day of hatch by skilled and experienced personnel, little measurable effect on early performance was observed during the first 6 wk of life. Decreased broiler performance is generally considered a sensitive indication of physical or psychogenic stress. Given the marked reduction in beak-inflicted trauma with beak trimming birds as they reach sexual maturity, these results suggest that when properly performed, neither of these beak-trimming methods causes sufficient physical or psychogenic stress to markedly affect early growth rate.

Sex differences in plasma corticosterone release in undisturbed chickens (Gallus gallus) in response to arginine vasotocin and corticotropin releasing hormone.

In birds, two neuropeptides, corticotropin releasing hormone (CRH) and arginine vasotocin (AVT), are major regulators of the hypothalamo-pituitary-adrenal axis (HPA) during the stress response. In birds, however, the relative efficacy of CRH and AVT to stimulate the HPA axis in males and females remains unknown. The purpose of this study was to determine the time course of CORT release following central CRH and AVT administration to male and female chickens. Chickens were fitted with a stainless steel cannula surgically implanted in the lateral ventricle and a catheter chronically inserted in the jugular vein. Birds were housed individually in cages behind a one-way glass partition and unnecessary noise was avoided during the sampling period. Each bird received a single 5.0microtracerebroventricular (ICV) injection of either saline (SAL), AVT (10 and 100pmol), or CRH (10 and 100pmol). Blood was sampled remotely every 15min for 2h and plasma CORT was determined by radioimmunoassay. There was a significant increase in plasma CORT concentration in males injected with 100pmol AVT beginning at 15min post-injection through 2h compared with SAL injected birds. In males, injection of 100pmol CRH was significantly more effective in releasing CORT compared to an equal molar concentration of AVT or SAL. In females, ICV injection of 100pmol AVT induced moderate increase in CORT levels. In contrast, 100pmol CRH significantly increased plasma CORT compared to SAL injected controls but the CORT response was nearly 50% less than that obtained in males.

Neurobiological basis of sensory perception: welfare implications of beak trimming.

The practice of beak trimming in the poultry industry occurs to prevent excessive body pecking, cannibalism, and to avoid feed wastage. To assess the welfare implications of the procedure, an emphasis of this paper has been placed on the anatomical structures that comprise the beak and mouth parts and a representation of the structures removed following beak trimming. Five animal welfare concerns regarding the procedure have been addressed, including the following: loss of normal beak function, short-term pain and temporary debilitation, tongue and nostril damage, neuromas and scar tissue, and long-term and phantom limb pain. Because all of the concerns involve the nervous system, the current knowledge of the avian somatosensory system was summarized. The critical components include touch, pain, and thermal receptors in the buccal cavity and bill, the trigeminal system, and neural projections mapped to the pallium (cortical-like tissue in the avian forebrain). At the present time, a need remains to continue the practice of beak trimming in the poultry industry to prevent head, feather, and vent pecking in some lines of birds. The procedure, however, should involve conservative trimming and be limited to young birds. Importantly, data show that removing 50% or less of the beak of chicks can prevent the formation of neuromas and allow regeneration of keratinized tissue to prevent deformed beaks and therefore positively affect the quality of life of birds during their lifetime.

Effects of Phos-Chek G75-F and Silv-Ex on red-winged blackbird (Agelaius phoeniceus) embryos.

Effects of field application levels of wildfire control chemicals, Phos-Chek G75-F (PC) and Silv-Ex (SE), were examined on red-winged blackbird (Agelaius phoeniceus) embryos. Embryos were more sensitive to PC and SE when eggs were immersed for 10s at an early developmental stage (days 3-5 of incubation) than at a later stage (days 6-9 of incubation). The LC(50) (concentration causing 50% mortality) for early stage embryos exposed to PC was 213.3g/L (slope=1.6; 95% confidence interval [CI]=129.1-326.1). The no observed effect concentration (NOEC) was below 135g PC/L, which caused a significant increase in embryonic mortality and represents the lowest field coverage level of 1gal/100feet(2). The LC(50) for early stage embryos exposed to SE was 19.8g/L (slope=1.5; 95% CI=11.7-52.2). Significant mortality was observed at 10g SE/L and marginal at 7.5g SE/L with an apparent NOEC around 5g SE/L. Neither chemical resulted in apparent developmental malformations.

The use of vitamin A-deficient diets and jugular vein ligation to increase intracranial pressure in chickens (Gallus gallus).

A technique was developed to record intracranial cerebrospinal fluid pressure (iCSFp) in chicks and mature chickens. Using that procedure, 2 methods were found to effect a significant elevation in intracranial pressure: 1) feeding a purified diet to roosters for 40 d containing 25% of the bird's requirement for vitamin A, and 2) ligating both jugular veins in birds. The purified diet significantly reduced plasma retinol levels from 1.080 to 0.046 microg/mL, and iCSFp was significantly elevated from 63.0 to 106.0 mmH2O (P < or = 0.05). Two limitations for using hypovitaminosis A diets were capillary fragility and a cisterna magna that did not develop to the size of that structure in birds of the same age fed control diets with adequate vitamin A content. The second procedure, a reversible surgical technique, showed that within 2.5 h from jugular vein ligation, intracranial pressure rose to 109.7 mmH2O, comparable with levels attained following feeding a vitamin A deficient diet to roosters. Bilateral clamping of the jugular veins overnight resulted in an elevation of iCSFp to 127 +/- 8.86 mmH2O. Results suggest that the chicken may be a useful animal model to investigate intracranial hypertension and its accompanying headaches known to occur in humans.

Attenuation of gonadal response to photostimulation following ablation of neurons in the lateral septal organ of chicks.

Many avian species in temperate zones respond to long photoperiods by showing recrudescence of gonads. Compelling evidence show that non-retinal, non-pineal photoreceptors exist in the avian brain. Within the ventral forebrain are specialized neurons that respond to light found in the medial portion of a circumventricular organ called the lateral septal organ (LSO). The objective of this study was to examine whether the integrity of the LSO was critical for rapid gonadal development in young male chicks placed under long day photostimulation. Birds were initially kept on a short photoperiod (LD 8:16) until 2 weeks of age, at which time bilateral electrolytic lesions were administered to the LSO. After surgery, birds were transferred to a long photoperiod (LD 16:8) and fed a chick starter diet containing 0.2% sulfamethazine (SMZ). The combination of a long day plus SMZ is known to stimulate sexual maturation in male chicks. In four separate experiments conducted to date, bilateral lesions directed to the LSO and lesions that missed and were placed caudal and ventral to the target resulted in a significant decrease in testes weight, compared to sham-operated controls (P<0.05). Results suggest that neurons in the LSO may be directly involved in responding to long photoperiods and stimulating gonadal development in broiler chicks.

Sulfamethazine advances puberty in male chicks by effecting a rapid increase in gonadotropins.

A sulfonamide, sulfamethazine (SMZ) has been shown to have a robust, progonadal effect. The mechanism of action of SMZ, however, is unknown. Our hypothesis is that the compound may act centrally and/or at the level of the pituitary. Four experiments were completed to test that hypothesis. Chicks exposed to a continuous photoperiod and fed a diet containing 0.2% SMZ showed an exponential increase in testes size. When 6 weeks of age (5 weeks on the SMZ diet), experimentals had testes weight nine times heavier than controls. Profiles for thyroid and gonadotropin plasma hormones suggested that T(3) was transiently lower in experimentals solely during the first week on treatment, while thyroxine levels were not different from controls. In contrast, luteinizing hormone (LH) and follicle stimulating hormone (FSH) were significantly elevated at the initial 1-week sampling point and remained elevated throughout the entire experiment. In a follow-up study, LH was found significantly higher than controls by 48 h after initially consuming the compound. When T(3) was added to the SMZ diet at 0.5 ppm, the progonadal effect of SMZ was attenuated. Importantly, chronic intake of T(3) delayed but did not block the stimulatory effect of SMZ for increasing plasma LH. We conclude that since one of the primary effects of SMZ is to increase rapidly plasma gonadotropins, data suggest the compound is acting at the level of the brain or pituitary to stimulate early gonadal development in chicks.

Neurobiology of molt in avian species.

Postnuptial molt is the major molt that occurs in most wild and domestic avian species each year. The process is much more than the replacement of feathers. Studies have shown that a significant increase in metabolic rate, increase in whole body protein synthesis, osteoporosis, loss of body fat, and a suppression of the immune system occur during this event of a bird's annual cycle. Several procedures have been developed to initiate feather replacement, and this review addresses hormonal and neuropeptide treatments that effected molt. The administration of thyroxine, progesterone, gonadotropin-releasing hormone agonist, and prolactin were examined. Of the four, the two most effective were thyroxine and prolactin administration. The neural modulators known to release thyroxine and prolactin, respectively, are thyroid hormone releasing hormone (TRH) and vasoactive intestinal polypeptide (VIP). To gain insight into the possible functions of VIP and TRH, the distribution of these neuromodulators and their terminal fields were reviewed in the avian brain. It was found that VIP-containing neurons and fibers identified a neural system in birds comparable to the visceral forebrain system (VFS) described in mammals. The VFS functions to regulate the balance of the autonomic nervous system. The autonomic nervous system and, therefore, the VFS have been proposed to regulate the many behavioral and physiological events in the annual cycle of a bird, including postnuptial molt. In contrast to VIP that has an extensive brain distribution throughout the forebrain and brainstem, TRH is relatively restrictive and has a main concentration in nerve cells in and about the paraventricular nucleus, a key neural component of the VFS. Due to the roll of TRH in regulating the thyroid axis and operating within the framework of the VFS, it is proposed that the peptide functions to shift the balance of the autonomic nervous system in the direction of the sympathetic nervous system. A shift toward the dominance of the sympathetic nervous system appears to be required during this phase of a bird's yearly cycle.

Effects of Phos-Chek G75-F and Silv-Ex on developing northern bobwhite quail (Colinus virginianus).

Eggs of nesting birds situated in peripheral areas serving as fire breaks are at risk of being sprayed with fire control chemicals. Acute toxicity tests were conducted by immersing northern bobwhite quail eggs for 10 s in different water-based concentrations of Silv-Ex(R) (SE), a foam-suppressant chemical, and Phos-Chek(R) G75-F (PC), a fire retardant chemical, on day 4 or day 11 of incubation. An attempt was made to relate the treatment concentrations to the actual field application levels. Mortality appeared higher in most groups exposed on day 11 than on day 4, suggesting that on day 11 the extensive chorioallantoic vascular network permitted greater uptake of chemical. Only 24-60% of the embryos survived to hatch at exposure concentrations of 202, 269, and 454 g/L PC when treated on incubation day 11. At higher concentrations including 681956, and 1211 g/L PC, the compound did not completely dissolve in water and clumped on eggshells, resulting in greater hatching success. Exposures to SE at 100 g/L on incubation day 11 did not significantly affect hatching success of embryos but did significantly reduce the percent hematocrit in blood compared with controls. Incubation day 11 exposure to 202 and 1211 g/L PC led to a significant increase in plasma aspartate aminotransferase, and day 4 exposure to 1211 g/L PC resulted in a significant increase in alanine aminotransferase. In addition to elevated liver enzymes, these treatments resulted in a decrease in the number of hepatocyte profiles (1211 g/L PC at day 4 and day 11) and an increase in hepatocyte size (202 and 1211 g/L PC at day 11) in hatchlings. A combination of SE and PC was synergistic (202 g/L PC and 50 g/L SE) at day 11 of incubation with respect to decreased hatching success and reduced bone lengths. However, lower concentrations of SE (10 g/L or 30 g/L) combined with 202 g/L of PC appeared antagonistic. This may be due to SE, as a surfactant, altering the ability of PC to penetrate the egg. Our results show fewer adverse effects following exposure to SE than to PC; therefore application of SE may be less harmful to breeding bird populations.

Increased proliferative activity and programmed cellular death in the turkey hen pituitary gland following interruption of incubation behavior.

Incubation behavior or broodiness in turkey hens is characterized by ovarian regression, hyperprolactinemia, and persistent nesting. Nest-deprivation of incubating turkey hens results in disruption of broodiness accompanied by a precipitous decline in plasma prolactin (PRL) concentrations. The objective of the present study is to examine cellular changes in the pituitary gland associated with nest-deprivation for 0, 1, 2, 3, 4, or 7 days. Bromodeoxyuridine (BrdU) was administered prior to kill to study proliferative activity. Pituitary tissue sections were immunostained using turkey growth hormone (GH) antibody, and/or chicken PRL peptide antibody, and BrdU antibody. Plasma PRL concentrations declined significantly following nest-deprivation for 1 or more days. The midsagittal pituitary area immunoreactive (ir) to GH was significantly increased while that of PRL was significantly decreased following nest-deprivation for 2 or more days. Terminal deoxy-UTP nick end labeling and PRL-immunostaining revealed an abundance of apoptotic nuclei in both cephalic and caudal lobes of the anterior pituitary gland, suggestive of programmed cellular death of lactotrophs in the pituitary gland of hens nest-deprived for 2 or more days. Mammosomatotrophs were abundant in hens nest-deprived on Day 0 but were absent in hens nest-deprived for 1 or more days. Proliferating (BrdU-ir) cells were significantly abundant in the pituitary cephalic and caudal lobes following nest-deprivation for 1 or more days but were absent on Day 0 or in laying hens. Dual-labeling studies indicated that most of the BrdU-ir nuclei in the caudal lobe were not colocalized in somatotrophs in hens nest-deprived for 1-4 days but did colocalize with GH following 7 days of nest-deprivation. In conclusion, nest-deprivation of incubating turkey hens results in 1) a precipitous decline in plasma PRL concentration, 2) programmed cell death of lactotrophs, 3) disappearance of mammosomatotrophs, 4) increased proliferative activity of pituitary cells, and 5) recruitment of somatotrophs arising primarily from mitosis of nonsomatotrophic cells.

Central nervous system regulation of gonadal development in the avian male.

Many avian species, particularly domesticated ones used for egg and meat production, are photoperiodic. Research conducted over the past several years has revealed the neural components, neurotransmitters, neuromodulators, and gonadotropins that play an important role in responding to changes in day length. An ultimate effect of the neuroendocrine response of birds to light is gonadal development and production of fertilized eggs and young for the next generation. The main purpose of this paper is to address the major neural systems that have been shown to affect reproductive function in males. Potential areas of research that would help elucidate the mechanism of neural activation of gonadal function are suggested.

Identification of growth-hormone- and prolactin-containing neurons within the avian brain.

Prolactin (PRL)- and growth-hormone (GH)-containing perikarya and fibers independent of the anterior pituitary gland have been reported to exist in the central nervous system of several mammalian species. The specific distributions of PRL- or GH-like neurons in the avian forebrain and midbrain, however, have not been reported. The objective of the study was to identify GH- and PRL-containing neurons in the hypothalamus and a few extrahypothalamic areas of two avian species. Brain and peripheral blood samples were collected from laying and broody turkey hens and ring doves. Broody turkey hens and doves had significantly higher plasma PRL concentrations compared with laying hens. Coronal brain sections were prepared and immunostained using anti-turkey GH and anti-chicken synthetic PRL antibodies. In turkey hens, the most dense GH-immunoreactive (ir) perikarya and fibers were found in hippocampus (Hp), periventricular hypothalamic nucleus, paraventricular nucleus, inferior hypothalamic nucleus, infundibular hypothalamic nucleus, medial and lateral septal area, and external zone of the median eminence (ME). In the ring dove, a similar pattern of distribution of GH-ir neurons was noticed at the brain sites listed above except that GH-ir fibers and granules were found only in the internal zone of ME and not in the external zone. In both turkeys and doves, the most immunoreactive PRL-ir perikarya and fibers were found in the medial and lateral septal area, Hp (turkey only), and bed nucleus of the stria terminalis pars magnocellularis. There were no apparent differences in the staining pattern of GH- or PRL-ir neurons between the laying and broody states in either species. However, the presence of GH-ir- and PRL-ir perikarya and fibers in several hypothalamic nuclei indicates that GH and PRL may influence parental behavior, food intake, autonomic nervous system function, and/or reproduction.

Interactions of Glucocorticoids, NPY and Hypothalamic Serotonin.

Neuropeptide Y (NPY), insulin, corticosterone (CORT) and 5-hydroxytryptamine (5-HT), or serotonin, are all involved in energy homeostasis. To show how they might interact, the effects of NPY on plasma insulin and on 5-HT metabolism in the paraventricular hypothalamic nucleus (PVN), the ventromedial hypothalamic nucleus (VMN), the arcuate nucleus (ARC), and the locus coeruleus (LC) were measured in separate groups of intact, adrenalectomized (ADX), or ADX and CORT replaced Sprague-Dawley rats. Fifteen minutes after ICV injection of NPY, no effect on 5-HT metabolism in the intact animal was observed. 5-HT and its metabolite, 5-HIAA, increased in the PVN as a result of ADX, an effect that was reversed with CORT replacement. NPY also reversed these effects. Similar effects of ADX, CORT replacement and NPY were noted with 5-HIAA in the ARC. This suggests a role for 5-HT in the attenuation of weight gain in ADX animals. NPY also caused an increase in plasma insulin in the CORT replaced animal, but not the intact animal. This was most likely due to an inability of CORT replaced animals to adjust their CORT levels in response to NPY.

Development of sexually dimorphic vasotocinergic system in the bed nucleus of stria terminalis in chickens.

The bed nucleus of stria terminalis (BnST) of the domestic fowl contains two groups of parvicellular vasotocinergic neurons that are sexually dimorphic. In adult cockerels, arginine vasotocin (AVT) synthesis is well expressed in the dorsolateral and ventromedial portions of the BnST, whereas in corresponding brain areas of hens, AVT synthesis is completely lacking. In the present study, in situ hybridization and immunocytochemical methods were used to compare the ontogeny of sexually dimorphic AVT gene expression in the BnST of male and female chickens from day 12 of embryonic development (E12) until the onset of sexual maturation. By E12, both parvicellular groups of AVT-immunoreactive (AVT-ir) perikarya in the developing BnST can be distinguished in some males, whereas in females their presence is questionable. A quantitative analysis, beginning at E14, showed that the parvicellular dorsolateral portion of the BnST of male embryos had more AVT perikarya compared with females. In contrast, no evident sex difference in distribution pattern and number of AVT mRNA containing neurons in this BnST portion was observable by in situ hybridization at E15. At E18, as well as on the first and second days posthatch (D1 and D2), no differences in the number of AVT synthesizing cells and intensity of immunoreactive staining in male versus female chickens were found. Between D2 and D7, the number of AVT-ir cells in the BnST declined rapidly in both sexes until it disappeared completely in females before D35. In males, another increase in sexually dimorphic AVT-ir cells and innervation of the lateral septum was associated with the onset of puberty and fully matched a pattern observed in adult fowls. These results demonstrate that the sexually dimorphic part of the AVT system undergoes sexual differentiation during early stages of ontogeny.

Neural sites and pathways regulating food intake in birds: a comparative analysis to mammalian systems.

The paper reviews hypotheses explaining the regulation of food intake in mammals that have addressed specific anatomical structures in the brain. An hypothesis, poikilostasis, is introduced to describe multiple, homeostatic states whereby the regulation of metabolism and feeding occur in birds. Examples are given for both wild and domestic avian species, illustrating dynamic shifts in homeostasis responsible for the changes in body weights that are seen during the course of an annual cycle or by a particular strain of bird. The following neural structures are reviewed as each has been shown to affect food intake in birds or in mammals: ventromedial hypothalamic nucleus (n.), lateral hypothalamic area, paraventricular hypothalamic n., n. tractus solitarius and area postrema, amygdala, parabrachial n., arcuate n. and bed n. of the stria terminalis. Two neural pathways are described which have been proposed to regulate feeding. The trigeminal sensorimotor pathway is the most complete neural pathway characterized for this behavior and encompasses the mechanics of pecking, grasping and mandibulating food particles from the tip of the bill to the back of the buccal cavity. A second pathway, the visceral forebrain system (VFS), affects feeding by regulating metabolism and the balance of the autonomic nervous system. Wild, migratory birds are shown to exhibit marked changes in body weight which are hypothesized to occur due to shifts in balance between the sympathetic and parasympathetic nervous systems. Domestic avian species, selected for a rapid growth rate, are shown to display a dominance of the parasympathetic nervous system. The VFS is the neural system proposed to effect poikilostasis by altering the steady state of the autonomic nervous system in aves and perhaps is applicable to other classes of vertebrates as well.