Central prolactin binding site densities change seasonally in an adult male passerine bird (Junco hyemalis).

Seasonal reproduction is common across temperate zone avian species. In these species, physiological and behavioral adaptations have evolved to change according to day length (i.e., seasonally) in order to maximize reproductive output. The hormone prolactin regulates many aspects of parental care, a critical component of reproductive success. It's secretion in birds has been shown to be under photoperiodic control, with the highest levels measured in the spring and summer months, when birds breed and show parental care. However, to date, no study has tested whether the densities of central prolactin binding sites vary seasonally, which may also account for prolactin's effect on parental care. To test this, we collected brains from free-ranging adult male dark-eyed juncos, Junco hyemalis, a biparental songbird, in the spring, summer, and fall, and used quantitative in vitro autoradiography to compare the densities of specific prolactin binding sites across 20 different brain regions. Prolactin binding sites were found in regions that regulate parental behavior in other avian species. During the summer, several hypothalamic regions that regulate parental care, including the preoptic area and tuberal nucleus, contained lower densities of prolactin binding sites, suggesting exposure to higher endogenous prolactin levels, than at other times. This observation is consistent with the fact that circulating prolactin is highest during summer, when males would be providing care to young. Overall, these data suggest that prolactin binding sites are relatively conserved in the avian brain and that central prolactin activity supports parental care efforts in juncos and other avian species.

Increased STAT5 signaling in the ring dove brain in response to prolactin administration and spontaneous elevations in prolactin during the breeding cycle.

Prolactin acts on target cells in the central nervous system (CNS) to stimulate behavioral changes associated with parental care in birds, but the signaling mechanisms that mediate these actions have not been characterized. In mammals, the Janus Kinase 2-Signal Transducer and Activator of Transcription 5 (JAK2-STAT5) signaling pathway mediates many of the actions of prolactin. To assess the importance of this pathway in prolactin-sensitive target cells in the avian brain, we measured changes in activated (phosphorylated) STAT5 (pSTAT5) in the forebrain of female ring doves sampled as plasma prolactin levels change during the breeding cycle and in prolactin-treated, non-breeding females. The anatomical distribution of cells exhibiting pSTAT5 immunoreactivity in dove brain closely paralleled the distribution of prolactin receptors in this species. The density of pSTAT5 immunoreactive (pSTAT5-ir) cells was highest in the preoptic area, the suprachiasmatic, paraventricular, and ventromedial hypothalamic nuclei, the lateral and tuberal hypothalamic regions, the lateral bed nucleus of the stria terminalis, and the lateral septum. Mean pSTAT5-ir cell densities in these eight brain areas were several fold higher in breeding females during late incubation/early post-hatching when plasma prolactin levels have been observed to peak than in non-breeding females or breeding females sampled at earlier stages when prolactin titers have been reported to be lower. Similar differences were observed between prolactin-treated and vehicle-treated females in all three of the forebrain regions that were compared. We conclude that JAK2-STAT5 signaling is strongly activated in response to prolactin stimulation in the ring dove brain and could potentially mediate some of the centrally-mediated behavioral effects of this hormone.

Courtship interactions stimulate rapid changes in GnRH synthesis in male ring doves.

Many birds and mammals show changes in the hypothalamo-pituitary-gonadal (HPG) axis in response to social or sexual interactions between breeding partners. While alterations in GnRH neuronal activity play an important role in stimulating these changes, it remains unclear if acute behaviorally-induced alterations in GnRH release are accompanied by parallel changes in GnRH synthesis. To investigate this relationship, we examined changes in the activity of GnRH neurons in the brains of male ring doves following brief periods of courtship interactions with females. Such interactions have been previously shown to increase plasma LH in courting male doves at 24 h, but not at 1 h, after pairing with females. In the first study, males allowed to court females for 2 h had 60% more cells that showed immunocytochemical labeling for GnRH-I in the preoptic area (POA) of the hypothalamus than did control males that remained isolated from females. To determine whether an increase in GnRH gene expression preceded this increase in GnRH immunoreactivity in the POA, changes in the number of cells with detectable GnRH-I mRNA in the POA were measured by in situ hybridization following a 1 h period of courtship interactions with females. In this second study, courting males exhibited 40% more cells with GnRH-I in this region than did isolated control males. GnRH-immunoreactive neurons in two other diencephalic regions failed to show these courtship-induced changes. Plasma LH was not elevated after 1 or 2 h of courtship. These results demonstrate that the release of GnRH-I in the POA that is presumably responsible for courtship-induced pituitary and gonadal activation is accompanied by a rapid increase in GnRH synthesis that occurs before plasma LH levels increase. We suggest that this increase in GnRH synthesis is necessary to support the extended period of HPG axis activation that is seen in this species during the 5-10 day period of courtship and nest building activity.

Patterns of fos-like immunoreactivity in the brains of parent ring doves (Streptopelia risoria) given tactile and nontactile exposure to their young.

Neuronal activation was examined by fos immunohistochemistry in ring doves (Streptopelia risoria) reunited with their young after overnight separation. In an initial study, squab-exposed parents showed more fos immunoreactivity (ir) in the preoptic area (POA) and lateral hypothalamus (LH) than squab-deprived parents. In a 2nd study, parents allowed free access to young and those separated from young by a wire mesh partition showed more fos-ir in the POA, LH, and lateral septum than box-exposed controls. Contact with young also increased fos-ir in the medial preoptic nucleus and bed nucleus of the stria terminalis, but noncontact exposure did not. Conversely, nontactile squab exposure stimulated more fos-ir in the POA than did free access to young, which suggests POA involvement in appetitive aspects of parenting.

Prolactin-induced parental hyperphagia in ring doves: are glucocorticoids involved?

Hyperphagia is a prominent component of the parental behavior repertoire in male and female ring doves and is necessary in order for parents to successfully provision their growing young. Although previous studies implicate both prolactin and the endogenous glucocorticoid, corticosterone, in parental hyperphagia, the functional interactions between these two hormones in regulating changes in feeding activity have not been characterized. These studies examined the possibility that prolactin's orexigenic effects are mediated through the increased secretion of corticosterone. Twice-daily intracerebroventricular (icv) injection of prolactin increased plasma corticosterone concentration in non-breeding doves of both sexes, with males exhibiting more pronounced effects than females. To further test the importance of glucocorticoid signaling in prolactin-induced feeding responses, changes in food intake were investigated in icv prolactin-treated, non-breeding doves following icv infusion of the glucocorticoid receptor antagonist RU38486 or propylene glycol vehicle. No attenuation of prolactin-induced hyperphagia was observed in either sex following co-administration of RU38486 at a dose shown previously to block dexamethasone-induced feeding in doves. These findings suggest that elevated corticosterone titers in blood may contribute to the hyperphagia observed in response to prolactin, but corticosterone signaling through a mammalian-type glucocorticoid receptor is not essential.

The role of the melanocortin system and the melanocortin-4 receptor in ring dove (Streptopelia risoria) feeding behavior.

The melanocortin-4 receptor (MC4-R) is an important mediator of the effects of two melanocortin system ligands, alpha melanocyte stimulating hormone (alpha-MSH) and agouti-related peptide (AGRP), on feeding behavior and energy balance in mammals. Although an avian homologue of the mammalian MC4-R has recently been identified, there is little information on the role of this receptor and the melanocortin system in avian feeding and body weight regulation. In these studies, we measured changes in feeding behavior in ring doves (Streptopelia risoria) following intracerebroventricular (i.c.v.) injection of various melanocortin receptor agonists and antagonists. The selective MC4-R antagonist HS014 elevated food intake within 4 h at all three doses tested (0.02, 0.2, and 2 nmol). A 1 nmol dose of the endogenous antagonist AGRP also stimulated feeding but only after a post-injection interval of 10 h. Surprisingly, the MC3-R and MC4-R antagonist SHU9119 not only failed to stimulate food intake at the same doses as HS014, but actually inhibited food intake at 8 h after injection. Whether this was due to toxicity effects or differences in the pharmacology of avian and mammalian melanocortin receptors remains to be determined. Food-deprived doves showed a fourfold increase in the number of AGRP-immunoreactive cells in the tuberal region of the hypothalamus and 5 ng of the MC3-R and MC4-R agonist MTII significantly attenuated the amount of food consumed by food-deprived birds that were allowed to re-feed. These data support a role for the melanocortin system and the melanocortin-4 receptor in the ring dove feeding behavior.

Glucocorticoids and parental hyperphagia in ring doves (Streptopelia risoria).

These studies explored the possibility that glucocorticoids promote parental care in ring doves by mediating, at least in part, the pronounced increase in food consumption that parent doves exhibit while provisioning their young. Plasma concentrations of the endogenous glucocorticoid corticosterone were found to be significantly higher in breeding females during the posthatching phase than during the incubation period. These differences were not observed in male breeding partners, but sex differences in daily activity rhythms are well documented in breeding doves, and blood sampling at different times of day would be required to adequately characterize the pattern of corticosterone in males during these breeding stages. In studies on nonbreeding doves, twice-daily intracerebroventricular (icv) injections of the synthetic glucocorticoid dexamethasone (DEX) increased food intake by 25-50% in both sexes, and further studies in males revealed that the increase was directly related to the dose of DEX administered. The highest dose of DEX given icv (1.0 microg/day) was not effective in stimulating feeding when given systemically, thereby suggesting that the hyperphagic action of DEX is exerted directly on the central nervous system. The icv infusion of the selective glucocorticoid receptor antagonist RU38486 blocked the hyperphagic effects of twice-daily icv injections of DEX in both sexes. Collectively, these data support the hypothesis that corticosterone contributes to the parental hyperphagia exhibited by breeding doves during the posthatching period. They also suggest that these orexigenic effects are mediated in part by CNS binding sites that resemble mammalian glucocorticoid receptors.