Cage-Free Pullets Minimally Affected by Stocking Density Stressors.

Management choices during the pullet phase can affect behavior, welfare, and health later in life, but few studies have evaluated the pullet phase, particularly in extensive housing systems. This study was a 2 × 2 factorial randomized complete block design (RCBD) with two strains and two stocking densities. The Lohmann LB-Lite and Lohmann LSL-Lite were housed on the floor at high-stocking density (619-670 cm2/bird) and low-stocking density (1249-1352 cm2/bird), which changed with age from 2 to 16 weeks of age (WOA). Bird-based measures of appearance, blood parameters, organ measurements, and production values were evaluated. Stocking density alone affected (p < 0.05) only relative bursal weight (% of body weight)-3.32% in the low-density versus 3.08% in the high-density group. High-stocking density was correlated with decreased uniformity (high-89.33 ± 0.24%; low-90.41 ± 0.24; p < 0.02) and worse feather coverage in the brown strain. High-stocking density was correlated with greater uniformity (High-90.39 ± 0.24%; Low-88.47 ± 0.24%; p < 0.001) and better feather coverage in the white strain. This study's feed conversion ratio (FCR) was improved by 0.07 in the low-stocking density for both strains. The remaining parameters were affected by strain and age only. Thus, while stocking density effects vary slightly depending on the strain used, cage-free pullets had limited negative effects at both the high and low-stocking densities tested in this study; there were few to no changes in the numerous bird-based welfare parameters tested.

Chronic Heat Stress Part 2: Increased Stress and Fear Responses in F1 Pekin Ducks Raised from Parents That Experienced Heat Stress.

The effects of HS on the welfare of poultry have been reported to have a transgenerational effect on phenotype plasticity. The goal of our experiment was to determine whether parental exposure to HS would impair the performance, HPA axis response, or behavior of their offspring. We treated adult drakes and hens (n = 80 ducks/treatment) at peak lay with HS or the control temperature for 3 weeks and incubated eggs collected from the last 3 days of the experiment. We utilized 76 ducklings/parental treatment group: control (CON-F1) and HS (HS-F1). Weekly data for body weights, body condition scores (BCSs), and novel object test (NOT) were collected. At 3 weeks of age, the ducks (n = 6/treatment) were subjected to adrenocorticotropic hormone (ACTH/cosyntropin, 0.0625 mg/kg) challenge or vehicle as the control. Blood samples were collected at 0, 1, 2, 3, and 4 h relative to treatment for serum glucocorticoid and heterophil-to-lymphocyte ratio (HLR) analyses. All injected birds were euthanized with pentobarbital on the second day relative to ACTH administration, and the spleen and bursa were removed and weighed immediately. Duck level analyses were completed using one- or two-way ANOVA as appropriate. BCSs were analyzed using a chi-squared test. The HS-F1 ducks had a lower hatch weight (p < 0.05) compared with the CON-F1 ducks but no significant difference in growth rates during the 5-week period. NOT (n = 4) analyses showed that the HS-F1 ducks had a greater fear response (p < 0.001) compared with the CON-F1 ducks. Similarly, an ACTH stimulation test showed that the HS-F1 ducks had significantly (p < 0.05) heightened corticosterone and HLR responses compared with the CON-F1 ducks. The HS-F1 ducks showed altered baseline and ACTH-stimulated levels of cortisol compared with the controls. Our data suggest that parental exposure to HS impacts the HPA response and fearfulness of the F1 generation in Pekin ducks.

Comparing different environmental enrichments for improving the welfare and walking ability of male turkeys.

This study investigated age-related changes in turkey welfare measures (wounds, feather quality (FQ), feather cleanliness, and footpad condition (FCON)) and walking ability (gait) as influenced by different types of environmental enrichment (EE). Tom turkeys (n = 420) were randomly assigned to: straw bale (S), platform (P), platform + straw bale (PS), pecking block (B), tunnel (T) or control (C; no enrichment) group. Welfare measures and gait were assessed at 8, 12, 16 and 19 wk and analyzed using PROC LOGISTIC with Firth bias-correction. Better wing FQ with age was observed in turkeys in S and T groups. Turkeys in the S group had better wing FQ at 16 (P = 0.028) and 19 wk (P = 0.011) vs. 8 wk. Wing FQ (P = 0.008) was better at 19 vs. 8 wk for T turkeys. FCON worsened over time for turkeys in all treatment groups except for the S group. FCON was worse at 19 vs.8 wk for P (P = 0.024), PS (P = 0.039), B (P = 0.011), T (P = 0.004) and C (P = 0.014) turkeys and was worse at 19 vs. 12 wk for B (P = 0.038), T (P = 0.015) and C (P = 0.045) turkeys. FCON was worse at 19 vs. 16 wk for T (P = 0.007) and C (P = 0.048) turkeys. FCON was also worse at 16 vs. 8 wk for B (P = 0.046) turkeys. Gait worsened with increasing age in all treatment groups. Gait was worse at 19 wk for S (P < 0.001), P (P < 0.001), PS (P < 0.001) and B turkeys (P < 0.001) vs. earlier ages, while gait in T (P < 0.001) and C turkeys (P < 0.001) worsened starting at 16 wk.

Catecholamine concentrations in duck eggs are impacted by hen exposure to heat stress.

Rapid "fight-or-flight" responses to stress are largely orchestrated by the catecholamines. Moreover, catecholamines and catecholamine precursors are widely recognized to act as interkingdom signaling molecules among host and microbiota, as well as to serve as chemotactic signals for bacterial foodborne pathogens. While albumen and yolk concentrations of glucocorticoids have received extensive attention as non-invasive indicators of hen response to stress, little is known regarding the impact of the hen's stress response on in ovo catecholamine and catecholamine precursor concentrations. The aim of the present study was to determine norepinephrine and L-dopa concentrations in albumen and yolk of eggs laid by hens maintained under normal or heat stress conditions. Norepinephrine and L-dopa concentrations were also measured in oviductal tissue. Breeder ducks (∼35 weeks/age) were kept under normal (22°C) conditions or subjected to cyclical heat stress (35°C day/29.5°C night) for 3 weeks. Eggs (n = 12 per timepoint/group) were collected on a weekly basis. Hens were sacrificed at baseline or after 3 weeks of heat stress for oviductal tissue collection. Albumen, yolk, and oviduct concentrations of norepinephrine and L-dopa were determined using ultra high-performance liquid chromatography with electrochemical detection. Norepinephrine and L-dopa were detected in oviductal tissue as well as egg albumen and yolk. Norepinephrine concentrations were elevated (p < 0.05) in the yolk of eggs laid by the heat stress group compared to those of the control group. Norepinephrine concentrations in albumen were elevated (p < 0.05) in the heat stress group compared to control group at week 2. L-dopa concentrations were not significantly affected (p > 0.05) by heat stress in albumen, yolk, or oviductal tissue. Together, the present study provides the first evidence of the stress neurohormone, norepinephrine, in duck eggs and identifies that hen exposure to heat stress can affect in ovo norepinephrine concentrations. These data highlight the potential utility of in ovo catecholamine concentrations as non-invasive measures of the hen's response to stress, as well as warrants future research into whether hen deposition of stress-related neurochemicals into the egg could serve as a chemotactic signal in the vertical transmission of foodborne pathogens.

Transportation Stress Increases Fos Immunoreactivity in the Paraventricular Nucleus, but Not in the Nucleus of the Hippocampal Commissure in the Pekin Duck, Anas platyrhynchos domesticus.

Commercial poultry undergo transportation during their life, and the effects of transportation can negatively impact poultry production and welfare. In order to maintain physiological homeostasis, the hypothalamic−pituitary−adrenal axis (HPA) works to respond to stressors. Previous studies by others have shown contradictory effects of transportation on corticosterone release. However, recent studies from our lab and by others have shown that cortisol may also be an important hormone in the avian HPA. The purpose of our current study was to determine the effects of transportation stress on the stimulation of brain nuclei that regulate the HPA in birds, and on glucocorticoid (GC) secretion. To test this hypothesis, we collected blood and brain samples from developer drakes and hens (N = 10 per sex/time point): 24 h prior to transportation, immediately after transportation, 24 h after transportation, and 1 week after transportation. Serum GC levels and fos immunocytochemistry (ICC) within the nucleus of the hippocampal commissure (NHpC) and paraventricular nucleus (PVN) were measured. Data were analyzed using a two-way ANOVA. Post hoc analysis was completed using a Fisher's PLSD with a p < 0.05 considered significant. We observed a sex difference (p < 0.05) in both corticosterone and cortisol secretion in Pekin ducks, although neither GC showed a significant increase in secretion associated with transportation. However, we did observe a significant (p < 0.05) increase in fos-like immunoreactivity for 24 h in the PVN, but not in the NHpC. Further studies are required to determine the specific role that GCs play in the avian stress response and the short-term stressors that could have long-term physiological effects on birds.

Effect of pre-hatch incubator lights on the ontogeny of CNS opsins and photoreceptors in the Pekin duck.

The Pekin duck is a valuable agricultural commodity globally and in the United States. Pekin ducks are seasonal breeders; they are sensitive to light and thus, research on the neuroendocrine and behavioral responses are needed to maximize production and to improve their welfare. There is compelling evidence that specific wavelengths of light may adversely alter the growth and welfare of meat (grow out) ducks. However, despite a birds' dependence upon light, in commercial poultry hatcheries, incubators almost exclusively hold eggs in the dark. Therefore, our objective was to determine the effects of lighting on the expression of retina photoreceptors (RPs) and deep brain photoreceptors (DBPs) during duck embryological development. Two groups of ducks were raised with and without light over 21 d from egg laying, embryonic day 0. Brain and retinal tissues were collected at embryonic days 3, 7, 11, 16, and 21 of a 24 d incubation period. qRT-PCR was performed on RPs (OPN1LW, OPN2SW, OPN1SW, MAFA, RHO, and RBP3) and the DBP OPN4M from retinal and brain samples, respectively. We find that the presence and absence of light during pre-hatch incubation, had no influence on the expression of any retinal photoreceptor. However, a late embryological increase in DBP OPN4M expression was observed. Taken together, the impact of light during pre-hatch incubation does not impact the overall post-hatch production. However, future directions should explore how OPN4M pre-hatch activation impacts Pekin duck post-hatch development and growth.

Serum trimethylamine-N-oxide and gut microbiome alterations are associated with cholesterol deposition in the liver of laying hens fed with rapeseed meal.

Sinapine derived from cruciferous plants could be converted into trimethylamine by intestinal microbiota. Its metabolite, trimethylamine N-oxide (TMAO), is closely linked to increased risk of cardiovascular disease and fat deposition in mammals. Hens fed with rapeseed meal (RSM) suffered from fatty liver hemorrhage syndrome (FLHS). This study was conducted to investigate whether RSM-induced fatty liver is due to TMAO via altering microbiota composition and diversity. At 33 weeks of age, 600 laying hens were randomly divided into 5 treatment groups, namely control and 14% RSM treatment groups (DY5, with 16.2% erucic acid [EA] and 74.66% glucosinolate [Gl] contents; MB1, with 3.50% EA and 43.23% Gl contents; DY6, with 6.7% EA and 22.67% Gl contents; XH3, with 44.60% EA and 132.83% Gl contents) for 8 weeks. Results revealed that 3 hens died due to liver hemorrhage after ingesting 14% RSM diet. The 14% RSM decreased serum low-density lipoprotein cholesterol (LDL-C) content (P < 0.01) while tended to increase serum TMAO content compared to the control group (P = 0.08). The 14% RSM diet increased red oil O optical density (P < 0.01), and increased total cholesterol (TC) and LDL-C content in the liver (P < 0.01, and P < 0.01, respectively). The 14% RSM decreased liver total bile acid (TBA) content compared to the control (P < 0.01). The DY6 had a higher TBA content in the liver than the XH3 (P < 0.01). The 14% RSM decreased mRNA abundance of liver X receptors alpha (LXR-α, P = 0.01), and increased mRNA abundance of sterol response element binding protein 2 (SREBP-2, P = 0.04). Results revealed that the in-feed RSM could alter richness and diversity of cecal microbiota compared to the control (P < 0.05). Liver TC content and serum TMAO showed a negative relationship with Proteobacteria and Actinobacteria (P = 0.04). In conclusion, 14% RSM increased liver TC and induced high liver score of FLHS, which was possibly associated with the altered cecal microbiota composition, increased serum TMAO levels and LXR-α and SREBP-2 expressions.

ICV galanin-like peptide stimulates non-contact erections but not touch-based erections in adult, sexually experienced male rats.

Galanin-like peptide (GALP) is a neuropeptide transcribed only within the arcuate nucleus of the hypothalamus and is thought to be a mediator between energetics and reproductive function. Intracerebroventricular (ICV) injection of GALP is known to have effects on feeding, and to significantly increase gonadotropin releasing hormone- (GnRH-) mediated luteinizing hormone (LH) secretion. Furthermore, ICV GALP is known to stimulate fos production in the medial pre-optic area (mPOA) and to a lesser extent, the paraventricular nucleus (PVN). ICV injection of 5.0nmol GALP profoundly stimulates male rat sexual behavior. It is not known if GALP's effects on sex behavior are due to an increase in appetitive or mechanical (erectile) aspects of male sexual behavior. To determine this, sexually experienced male rats were cannulated in the lateral ventricle and injected with 5.0nmol GALP or vehicle. Immediately after injections, male rats were placed in an arena connected to a second arena via a tube with a fan. The second arena contained a steroid-primed female and her bedding. The male rat had olfactory but not visual or tactile contact with the female. We analyzed the amount of time the male rats spent investigating the air intake and the number of non-contact erections (NCEs) in a 30minute test. ICV GALP significantly (p<0.05) increased both the amount of time of olfactory investigations and NCEs compared to vehicle. In a second set of animals, we tested if ICV GALP could stimulate touch-based erections. GALP had no significant effect on touch-based erections compared to vehicle. These data suggest that GALP's activation of fos within the mPOA is indicative of its action to stimulate the appetitive aspects of male sexual behavior.

Injections of Galanin-Like Peptide directly into the nucleus of the tractus solitarius (NTS) reduces food intake and body weight but increases metabolic rate and plasma leptin.

Galanin-Like Peptide (GALP) is a hypothalamic neuromediator of metabolism and reproduction. GALP is known to stimulate reproduction and alter food intake and body weight in multiple species. The regulation of body weight involves control of both energy intake and energy expenditure. Since GALP is known to alter food intake - possibly via the autonomic nervous system - we first hypothesized that GALP would increase metabolic rate. First, male Sprague-Dawley rats were implanted with intracerebroventricular (ICV) cannulae and abdominal radiotelemetry temperature transmitters. Following ICV injection with either 5nmol GALP or vehicle, the oxygen consumption of each rat was monitored for 8h. Food intake, core temperature, and general motor activity were monitored for 24h. GALP significantly increased oxygen consumption, an indirect estimator of metabolic rate, without having any significant effect on motor activity. Compared to controls, GALP increased core body temperature during the photophase and reduced food intake over the 24h period following injection. ICV GALP also increased plasma levels of luteinizing hormone (LH). A second group of male Sprague-Dawley rats were implanted with abdominal transmitters and given injections of GALP directly into the nucleus of the tractus solitarius (NTS). These injections resulted in a significant reduction in food intake, and a significant increase in both oxygen consumption and core body temperature compared to vehicle injections. Direct injections of GALP into the NTS compared to vehicle also resulted in a significant increase in plasma leptin levels, but not LH levels. GALP appears to increase energy expenditure in addition to decreasing energy input by actions within the NTS and thus may play an important role in the hypothalamic regulation of body weight.

Characterization of Gut Microbiome Dynamics in Developing Pekin Ducks and Impact of Management System.

Little to no research has been conducted on the gut microbiome of the Pekin duck, yet over 24.5 million ducks are raised for human consumption each year in the United States alone. Knowledge of the microbiome could lead to an understanding of the effects of growing conditions such as the use of prebiotics, probiotics, and enzymes in feeding practices, the use of antibiotics, and the sources of pathogenic bacteria in diseased ducks. In order to characterize changes in the caecal microbiome that occur as ducks develop through a typical industry grow-out period, a 16S rRNA community analysis of caecal contents collected over a 6-week period was conducted using a next generation sequencing approach. Transitions in the composition of the caecal microbiome occurred throughout the lifespan, with a large shift during days 4 through 10 posthatch. Two major phyla of bacteria were found to be present within the caeca of aviary raised ducks, with the relative abundance of each phylum varying by age of the duck. Proteobacteria is dominant for the first 3 days of age, and Firmicutes increases and dominates beginning at day 4. Barn raised ducks contained a significant population of Bacteroidetes in addition to Proteobacteria and Firmicutes at later developmental time points, though this phylum was absent in aviary raised ducks. Genera containing pathogens of anseriformes most often found in industry settings were either absent or found as normal parts of the caecal microbial populations. The high level differences in phylum abundance highlight the importance of well-designed sampling strategies for microbiome based studies. Results showed clear distinctions between Pekin Duck caecal contents and those of Broiler Chickens and Turkey in a qualitative comparison. These data provide a reference point for studies of the Pekin Duck through industry grow-out ages, provide a foundation for understanding the types of bacteria that promote health, and may lead to improved methods to increase yields and decrease instances of disease in agricultural production processes.

Alarin 6-25Cys antagonizes alarin-specific effects on food intake and luteinizing hormone secretion.

Previous data from our labs and from others have demonstrated that intracerebroventricular (ICV) injection of alarin has orexigenic activity and significantly increases plasma luteinizing hormone (LH) secretion in a gonadotropin-releasing hormone (GnRH) dependent manner. The purpose of the current experiments was to determine if the amino acids at the amino-terminal end of the alarin peptide are critical for alarin's effects on reproductive and feeding systems. First, we injected male mice ICV with full-length alarin (Ala1-25) or peptide fragments missing residues at the amino-terminal end (Ala3-25 or Ala6-25 Cys). Neither peptide fragment alone, significantly increased food intake in male mice compared to controls. Second, ICV injection of Ala1-25, but not Ala3-25, significantly (p < 0.01) increased GnRH-mediated LH secretion. Surprisingly, Ala6-25 Cys significantly (p < 0.05) inhibited plasma LH secretion and inhibited Ala1-25 actions. In conclusion, elimination of the first five amino acids of alarin not only abolishes the biological activity of alarin, but becomes an antagonist to alarin-specific effects. Furthermore, Ala6-25 Cys seems to act as a specific antagonist to putative alarin receptors and therefore may be an important tool in identifying alarin-specific receptors.

Gonadotropin-inhibitory hormone is a hypothalamic peptide that provides a molecular switch between reproduction and feeding.

OBJECTIVE: Gonadotropin-inhibitory hormone (GnIH)-3 is a neuropeptide that plays a major role in the regulation of reproduction and feeding in mammals. MATERIALS AND METHODS: We measured endocrine and behavioural parameters of reproduction in sheep, and sexual behaviour in sheep, mice and cynomolgus monkeys. In addition, GnIH gene expression (in situ hybridization) was examined in ewes, and effects of GnIH-3 on food intake and energy expenditure were measured in various species. GnIH-3 was infused (i.v.) into ewes after an i.m. injection of estradiol benzoate to determine whether the peptide blocks the surge in luteinizing hormone (LH) secretion. RESULTS: GnIH gene expression was reduced in the preovulatory period in ewes. Infusion (i.v.) of GnIH-3 blocked the estrogen-induced LH surge (in ewes). Intracerebroventricular infusion had no effect on female or male sexual behaviour in each of the three species, but increased food intake. There were no effects on energy expenditure in sheep or rats. GnIH increased fos protein (immunohistochemistry) was seen in orexigenic neurons (in sheep and rats), but also in anorexigenic neurons (in sheep). CONCLUSIONS: GnIH-3 reduces reproductive hormone levels and increases food intake in mammals without reducing energy expenditure. There is minimal effect on reproductive behaviour. The dual effect on reproduction and feeding suggests that GnIH-3 provides a molecular switch between these two functions. Blockade of the positive feedback effect of estrogen with parenteral infusion indicates that this peptide may have utility as a blocker of reproductive function in mammals.

Effects of alarin on food intake, body weight and luteinizing hormone secretion in male mice.

Alarin is a member of the galanin family of neuropeptides that includes galanin and galanin-like peptide (GALP). Alarin is an alternate transcript of the GALP gene and is expressed in the brain and periphery. Recently, it was shown in male rats that alarin is an orexigenic peptide that also regulates reproductive hormone secretion. We hypothesized that alarin would also have similar central effects on feeding and luteinizing hormone (LH) secretion in mice as observed in rats. To test this hypothesis, we treated male mice with alarin intracerebroventricularly (i.c.v.) and measured its effects on food intake, body weight, body temperature, LH secretion, and Fos induction. We observed that i.c.v. injection of 1.0 nmol alarin significantly increased immediate food intake (p<0.01) from 30 to 120 min post-injection and relative body weight (p<0.05) after 24 h. Alarin had no effect on body temperature compared to controls. Alarin increased LH levels in male mice, an effect that was dependent on gonadotropin-Releasing-Hormone (GnRH) signaling. Furthermore, alarin-stimulated Fos immunoreactivity was observed in diencephalic nuclei, including the hypothalamic dorsomedial nucleus and the bed nucleus of the stria terminalis. Our studies demonstrated that alarin, like other members of the galanin peptide family, is a neuromediator of food intake and reproductive hormone secretion in male mice.

Galanin-like peptide (GALP) is a hypothalamic regulator of energy homeostasis and reproduction.

Galanin-like peptide (GALP) was discovered in 1999 in the porcine hypothalamus and was found to be a 60 amino acid neuropeptide. GALP shares sequence homology to galanin (1-13) in position 9-21 and can bind to, as well as activate, the three galanin receptor subtypes (GalR1-3). GALP-expressing cells are limited, and are mainly found in the arcuate nucleus of the hypothalamus (ARC) and the posterior pituitary. GALP-positive neurons in the ARC project to several brain regions where they appear to make contact with multiple neuromodulators. These neuromodulators are involved in the regulation of energy homeostasis and reproduction, anatomical evidence that suggests a role for GALP in these physiological functions. In support of this idea, GALP gene expression is regulated by several factors that reflect metabolic state including the metabolic hormones leptin and insulin, thyroid hormones, and blood glucose. Considerable evidence now exists to support the hypothesis that GALP has a role in the regulation of energy homeostasis and reproduction; and, that GALP's role may be independent of the known galanin receptors. In this review, we (1) provide an overview of the distribution of GALP, and discuss the potential relationship between GALP and other neuromodulators of energy homeostasis and reproduction, (2) discuss the metabolic factors that regulate GALP expression, (3) review the evidence for the role of GALP in energy homeostasis and reproduction, (4) discuss the potential downstream mediators and mechanisms underlying GALP's effects, and (5) discuss the possibility that GALP may mediate its effects via an as yet unidentified GALP-specific receptor.

Alarin stimulates food intake in male rats and LH secretion in castrated male rats.

Alarin is a newly identified member of the galanin family of neuropeptides that includes galanin-like peptide (GALP) and galanin. Alarin was discovered as an alternate transcript of the GALP gene in neuroblastoma cells, and subsequently alarin mRNA was detected in the brain of rodents. GALP and galanin are important central regulators of both feeding and reproductive behavior. We hypothesized, that, as a member of the galanin family of peptides, alarin would also have central effects on feeding and reproduction. To test this hypothesis, we treated male rats with alarin intracerebroventricularly (i.c.v.) and measured its effects on food intake and energy homeostasis as well as sexual behavior and luteinizing hormone (LH) secretion. We observed that i.c.v. injection of 1.0 nmol alarin significantly increased food intake (p<0.01) and body weight (p<0.05). Alarin did not affect sexual behavior in male rats; however, alarin did significantly (p<0.01) increase LH levels in castrated, but not intact, male rats. Alarin immunoreactive cell bodies were detected within the locus coeruleus and locus subcoeruleus of the midbrain, which is a brainstem nucleus involved in coordinating many physiological activities, including food intake and reproduction. Lastly, alarin stimulated Fos induction in hypothalamic nuclei, such as the paraventricular nucleus and the nucleus of the tractus solitarious. Our studies demonstrate that alarin, like other members of the galanin family, is a neuromediator of food intake and body weight.

Neurons that co-localize aromatase- and kisspeptin-like immunoreactivity may regulate the HPG axis of the Mallard drake (Anas platyrhynchos).

Kisspeptin is a potent regulator of the hypothalamo-pituitary-gonadal axis. The activation of several vernal and pubertal behaviors involves the action of locally synthesized estradiol by hypothalamic aromatase-expressing neurons. Little is known about kisspeptin in non-mammalian systems, and its interaction with aromatase remains unexamined. The Mallard drake is a seasonal breeder and an excellent model for studying the neural mechanisms that regulate the HPG. The goals of these studies were to determine (a) if and how kisspeptin regulates the drake HPG, (b) if kisspeptin and aromatase are expressed in the Mallard brain, and (c) if kisspeptin is co-localized or in apposition with, aromatase- and gonadotropin hormone releasing hormone (GnRH) positive neurons. Central kisspeptin administration increased plasma luteinizing hormone, an effect blocked by pretreatment with the GnRH antagonist, acyline, suggesting a conservation of kisspeptin function and mechanism of action in birds and mammals. The distribution of kisspeptin in the mallard brain was examined with immunocytochemistry (ICC). Neurons that express kisspeptin-like immunoreactive (ir) protein were observed in the medial preoptic nucleus (POM) and in ir fibers throughout the drake brain. Virtually all POM kisspeptin-ir soma also expressed aromatase-ir, suggesting that autocrine mechanisms may predominate in the interaction between steroid provision and kisspeptin expression. No co-localization was observed between KP-ir and GnRH-ir, although both were easily detected in close-proximity in the tuberoinfundibular area. Taken together, these data suggest that in the drake, estradiol synthesized by aromatase and kisspeptin co-expressing POM neurons may regulate the HPG via an effect on GnRH secretion.

Seasonal changes in androgen receptor mRNA in the brain of the white-crowned sparrow.

In songbirds, neurons that regulate learned song behavior undergo extensive seasonal plasticity in their number and size in relation to the bird's reproductive status. Seasonal plasticity of these brain regions is primarily regulated by changes in circulating concentrations of testosterone. Androgen receptors are present in all of the major song nuclei, but it is unknown whether levels of androgen receptor mRNA in the telencephalic song regions HVC, the robust nucleus of the arcopallium, and the lateral magnocellular nucleus of the anterior nidopallium change as a function of season in white-crowned sparrows. To determine whether seasonal changes in levels of androgen receptor mRNA are specific to the song control system, we also measured levels of androgen receptor mRNA in a limbic nucleus, the lateral division of the bed nucleus of the stria terminalis (the lateral division of the bed nucleus of the stria terminalis). We found that levels of androgen receptor mRNA were higher in HVC and the lateral division of the bed nucleus of the stria terminalis of birds in the breeding condition compared with the nonbreeding condition; however, we observed no seasonal differences in levels of androgen receptor mRNA in either the robust nucleus of the arcopallium or the lateral magnocellular nucleus of the anterior nidopallium. These results are consistent with previous observations that seasonal plasticity of the song nuclei results from testosterone acting directly on HVC, which then exerts transsynaptic trophic effects on its efferent targets. The seasonal change in the expression of androgen receptor in HVC may be one component of the cellular mechanisms underlying androgenic effects on seasonal plasticity of the song control nuclei.

Galanin-like peptide: neural regulator of energy homeostasis and reproduction.

Galanin-like peptide (GALP) was discovered in 1999 in the porcine hypothalamus and was found to be a 60 amino-acid neuropeptide. GALP shares sequence homology to galanin (1-13) in position 9-21 and can bind to and activate the three galanin receptor subtypes (GalR1-3). GALP-expressing cells are limited, and are mainly found in the arcuate nucleus of the hypothalamus (ARC) and the posterior pituitary. GALP-positive neurons in the ARC, project to several brain regions where they appear to make contact with several neuromodulators that are involved in the regulation of energy homeostasis and reproduction, anatomical evidence that suggests a role for GALP in these physiological functions. In support of this idea, GALP gene expression is regulated by several factors that reflect metabolic state including the metabolic hormones leptin and insulin, thyroid hormones, and blood glucose. Considerable evidence now exists to support the hypothesis that GALP has a role in the regulation of energy homeostasis and reproduction; and, that that GALP's role may be independent of the known galanin receptors. In this chapter we (1) provide an overview of the distribution of GALP, and discuss the potential relationship between GALP and other neuromodulators of energy homeostasis and reproduction, (2) discuss the metabolic factors that regulate GALP expression, (3) review the evidence for the role of GALP in energy homeostasis and reproduction, (4) discuss the potential downstream mediators and mechanisms underlying GALP's effects, and (5) discuss the possibility that GALP may mediate it's effects via an as yet unidentified GALP-specific receptor.