Sex-specific behavioral and physiological changes during single parenting in a biparental species, Columba livia.

Many species exhibit biparental care to maximize fitness. When a partner is lost, the surviving partner may alter their behavior to compensate offspring. Whether both sexes use the same physiological mechanisms to manifest their change in behavior remains elusive. We investigated behaviors and mechanisms associated with the alteration of parental care post-partner removal in a biparental avian species, the rock dove (Columba livia). We hypothesized that rock dove single parents experience sex-biased changes in neural genomic transcription and reproductive behaviors, and these changes are related to chick development. We manipulated parental partner presence and measured parental attendance, offspring growth, gene expression of glucocorticoid receptors (GR) and mineralocorticoid receptors (MR) in the pituitary, and GR, MR, and estrogen receptor beta (ER-ß) in the hypothalamus. We also measured circulating plasma concentrations of the stress-associated hormone corticosterone and the parental care-associated hormone prolactin. We also quantified prolactin gene (PRL) expression changes in the pituitary, as well as prolactin receptor (PRLR) expression in the hypothalamus and pituitary. We found that single mothers and fathers maintained similar provisioning levels as paired parents, but spent less cumulative time brooding chicks. Chicks of single parents were smaller than paired-parented chicks after three days post-hatch. Mothers in both treatment groups experienced higher expression of hypothalamic GR as compared to fathers. Single parents experienced lower PRL gene expression in the pituitary as compared to paired parents. No significant differences were found for the circulating hormones or other genes listed.

Prolactin promotes parental responses and alters reproductive axis gene expression, but not courtship behaviors, in both sexes of a biparental bird.

Prolactin, a hormone involved in vertebrate parental care, is hypothesized to inhibit reproductive hypothalamic-pituitary-gonadal (HPG) axis activity during parenting, thus maintaining investment in the current brood as opposed to new reproductive efforts. While prolactin underlies many parental behaviors in birds, its effects on other reproductive behaviors, such as courtship, remain unstudied. How prolactin affects neuropeptide and hormone receptor expression across the avian HPG axis also remains unknown. To address these questions, we administered ovine prolactin (oPRL) or a vehicle control to both sexes in experienced pairs of the biparental rock dove (Columba livia), after nest removal at the end of incubation. We found that oPRL promoted parental responses to novel chicks and stimulated crop growth compared to controls, consistent with other studies. However, we found that neither courtship behaviors, copulation rates nor pair maintenance differed with oPRL treatment. Across the HPG, we found oPRL had little effect on gene expression in hypothalamic nuclei, but increased expression of FSHB and hypothalamic hormone receptor genes in the pituitary. In the gonads, oPRL increased testes size and gonadotropin receptor expression, but did not affect ovarian state or small white follicle gene expression. However, the oviducts of oPRL-treated females were smaller and had lower estrogen receptor expression compared with controls. Our results highlight that some species, especially those that show multiple brooding, may continue to express mating behavior despite elevated prolactin. Thus, mechanisms may exist for prolactin to promote investment in parental care without concurrent inhibition of reproductive function or HPG axis activity.

Prior parental experience attenuates hormonal stress responses and alters hippocampal glucocorticoid receptors in biparental rock doves.

In the face of challenges, animals must balance investments in reproductive effort versus their own survival. Physiologically, this trade-off may be mediated by glucocorticoid release by the hypothalamic-pituitary-adrenal axis and prolactin release from the pituitary to maintain parental care. The degree to which animals react to and recover from stressors likely affects maintenance of parental behavior and, ultimately, fitness. However, less is known about how gaining parental experience may alter hormonal stress responses and their underlying neuroendocrine mechanisms. To address this gap, we measured the corticosterone (CORT) and prolactin (PRL) stress response in individuals of both sexes of the biparental rock dove (Columba livia) that had never raised chicks versus birds that had fledged at least one chick. We measured both CORT and PRL at baseline and after an acute stressor (30 min restraint). We also measured negative feedback ability by administering dexamethasone, a synthetic glucocorticoid that suppresses CORT release, and measured CORT and PRL after 60 min. All hormones were measured when birds were not actively nesting to assess whether effects of parental experience extend beyond the breeding bout. Experienced birds had lower stress-induced and negative-feedback CORT, and higher stress-induced PRL than inexperienced birds. In a separate experiment, we measured glucocorticoid receptor subtype expression in the hippocampus, a key site of negative feedback regulation. Experienced birds showed higher glucocorticoid receptor expression than inexperienced controls, which may mediate their ability to attenuate CORT release. Together, these results shed light on potential mechanisms by which gaining experience may improve parental performance and fitness.

Prolactin and prolactin receptor expression in the HPG axis and crop during parental care in both sexes of a biparental bird (Columba livia).

During breeding, multiple circulating hormones, including prolactin, facilitate reproductive transitions in species that exhibit parental care. Prolactin underlies parental behaviors and related physiological changes across many vertebrates, including birds and mammals. While circulating prolactin levels often fluctuate across breeding, less is known about how relevant target tissues vary in their prolactin responsiveness via prolactin receptor (PRLR) expression. Recent studies have also investigated prolactin (PRL) gene expression outside of the pituitary (i.e., extra-pituitary PRL), but how PRL gene expression varies during parental care in non-pituitary tissue (e.g., hypothalamus, gonads) remains largely unknown. Further, it is unclear if and how tissue-specific PRL and PRLR vary between the sexes during biparental care. To address this, we measured PRL and PRLR gene expression in tissues relevant to parental care, the endocrine reproductive hypothalamic-pituitary- gonadal (HPG) axis and the crop (a tissue with a similar function as the mammalian mammary gland), across various reproductive stages in both sexes of a biparental bird, the rock dove (Columba livia). We also assessed how these genes responded to changes in offspring presence by adding chicks mid-incubation, simulating an early hatch when prolactin levels were still moderately low. We found that pituitary PRL expression showed similar increases as plasma prolactin levels, and detected extra-pituitary PRL in the hypothalamus, gonads and crop. Hypothalamic and gonadal PRLR expression also changed as birds began incubation. Crop PRLR expression correlated with plasma prolactin, peaking when chicks hatched. In response to replacing eggs with a novel chick mid-incubation, hypothalamic and gonadal PRL and PRLR gene expression differed significantly compared to mid-incubation controls, even when plasma prolactin levels did not differ. We also found sex differences in PRL and PRLR that suggest gene expression may allow males to compensate for lower levels in prolactin by upregulating PRLR in all tissues. Overall, this study advances our understanding of how tissue-specific changes in responsiveness to parental hormones may differ across key reproductive transitions, in response to offspring cues, and between the sexes.

Stress-mediated convergence of splicing landscapes in male and female rock doves.

BACKGROUND: The process of alternative splicing provides a unique mechanism by which eukaryotes are able to produce numerous protein products from the same gene. Heightened variability in the proteome has been thought to potentiate increased behavioral complexity and response flexibility to environmental stimuli, thus contributing to more refined traits on which natural and sexual selection can act. While it has been long known that various forms of environmental stress can negatively affect sexual behavior and reproduction, we know little of how stress can affect the alternative splicing associated with these events, and less still about how splicing may differ between sexes. Using the model of the rock dove (Columba livia), our team previously uncovered sexual dimorphism in the basal and stress-responsive gene transcription of a biological system necessary for facilitating sexual behavior and reproduction, the hypothalamic-pituitary-gonadal (HPG) axis. In this study, we delve further into understanding the mechanistic underpinnings of how changes in the environment can affect reproduction by testing the alternative splicing response of the HPG axis to an external stressor in both sexes. RESULTS: This study reveals dramatic baseline differences in HPG alternative splicing between males and females. However, after subjecting subjects to a restraint stress paradigm, we found a significant reduction in these differences between the sexes. In both stress and control treatments, we identified a higher incidence of splicing activity in the pituitary in both sexes as compared to other tissues. Of these splicing events, the core exon event is the most abundant form of splicing and more frequently occurs in the coding regions of the gene. Overall, we observed less splicing activity in the 3'UTR (untranslated region) end of transcripts than the 5'UTR or coding regions. CONCLUSIONS: Our results provide vital new insight into sex-specific aspects of the stress response on the HPG axis at an unprecedented proximate level. Males and females uniquely respond to stress, yet exhibit splicing patterns suggesting a convergent, optimal splicing landscape for stress response. This information has the potential to inform evolutionary theory as well as the development of highly-specific drug targets for stress-induced reproductive dysfunction.

Captivity alters neuroendocrine regulators of stress and reproduction in the hypothalamus in response to acute stress.

Wild animals are brought into captivity for many reasons. However, unlike laboratory-bred animals, wild caught animals often respond to the dramatic shift in their environment with physiological changes in the stress and reproductive pathways. Using wild-caught male and female house sparrows (Passer domesticus) we examined how time in captivity affects the expression of reproductive and stress-associated genes in the brain, specifically, the hypothalamus. We quantified relative mRNA expression of a neurohormone involved in the stress response (corticotropin releasing hormone [CRH]), a hypothalamic inhibitor of reproduction (gonadotropin inhibitory hormone [GnIH]), and the glucocorticoid receptor (GR), which is important in terminating the stress response. To understand potential shifts at the cellular level, we also examined the presence of hypothalamic GnIH (GnIH-ir) using immunohistochemistry. We hypothesized that expression of these genes and the abundance of cells immunoreactive for GnIH would change in response to time in captivity as compared to free-living individuals. We found that GR mRNA expression and GnIH-ir cell abundance increased after 24 and 45 days in captivity, as compared to wild-caught birds. At 66 days in captivity, GR expression and GnIH cell abundance did not differ from wild-caught birds, suggesting birds had acclimated to captivity. Evaluation of CRH and GnIH mRNA expression yielded similar trends, though they were not statistically significant. In addition, although neuroendocrine factors appeared to acclimate to captivity, a previous study indicated that corticosterone release and immune responses of these same birds did not acclimate to captivity, suggesting that neuroendocrine endpoints may adapt more rapidly to captivity than downstream physiological measures. These data expand our understanding of the physiological shifts occurring when wild animals are brought into captivity.

Sex-biased transcriptomic response of the reproductive axis to stress.

Stress is a well-known cause of reproductive dysfunction in many species, including birds, rodents, and humans, though males and females may respond differently. A powerful way to investigate how stress affects reproduction is by examining its effects on a biological system essential for regulating reproduction, the hypothalamic-pituitary-gonadal (HPG) axis. Often this is done by observing how a stressor affects the amount of glucocorticoids, such as cortisol or corticosterone, circulating in the blood and their relationship with a handful of known HPG-producing reproductive hormones, like testosterone and estradiol. Until now, we have lacked a full understanding of how stress affects all genomic activity of the HPG axis and how this might differ between the sexes. We leveraged a highly replicated and sex-balanced experimental approach to test how male and female rock doves (Columba livia) respond to restraint stress at the level of their transcriptome. Females exhibit increased genomic responsiveness to stress at all levels of their HPG axis as compared to males, and these responsive genes are mostly unique to females. Reasons for this may be due to fluctuations in the female endocrine environment over the reproductive cycle and/or their evolutionary history, including parental investment and the potential for maternal effects. Direct links between genome to phenome cause and effect cannot be ascertained at this stage; however, the data we report provide a vital genomic foundation on which sex-specific reproductive dysfunction and adaptation in the face of stress can be further experimentally studied, as well as novel gene targets for genetic intervention and therapy investigations.

An integrative overview of the role of gonadotropin-inhibitory hormone in behavior: applying Tinbergen's four questions.

The integration of various fields of investigation is of key importance to fully comprehending endocrine function. Here, I enact the theoretical framework of Nikolaas Tinbergen's four questions for understanding behavior to help bridge the wide gap that exists between our relatively reductionist molecular knowledge of a particular neurohormone, gonadotropin-inhibitory hormone (GnIH), and its place in animal behavior. Hypothalamic GnIH, upon its discovery in 2000, was so named because of its inhibitory effect on the release of the gonadotropins, luteinizing hormone (LH) and follicle stimulating hormone (FSH), from the pituitary. Because gonadotropins are necessary for reproduction, this finding stimulated questions about the functional significance of GnIH in reproduction and sexual behavior. After over a decade of research, invaluable knowledge has been gained regarding the mechanistic attributes of GnIH (mammalian homolog, RFamide-related peptide (RFRP)) in a variety of vertebrate species. However, many questions remain regarding the effect of the environment on GnIH and the subsequent effects of GnIH on behavior. I review the role of GnIH in shaping behavior using the framework of Tinbergen's four questions of mechanism, ontogeny, function and phylogeny. The studies I review were conducted in various species of mammals, birds, and in one species of fish. Because GnIH can play a role in mediating behaviors such as those important for reproduction, sociality, feeding, and the stress response in a variety of species, an integrative approach to the study of GnIH will help provide a multipronged schema for answering questions of GnIH function. By using the framework highlighted by Tinbergen's four questions, we will deepen and enhance our knowledge of the role of hormones in behavior from the point of view of the mechanisms involved.

Season- and context-dependent sex differences in melatonin receptor activity in a forebrain song control nucleus.

There are dense populations of melatonin receptors in large areas of the songbird brain, in particular in the visual system and the song control system. Melatonin has therefore been implicated in neuroplasticity of the song control system. Previously we demonstrated large changes in activity of melatonin receptor in Area X, a forebrain song control nucleus involved in song learning and production. In a laboratory environment, melatonin receptor activity was down-regulated in male and female European starlings during photostimulation (a simulated breeding season). The functional significance of this large change in Area X is unclear, so we sought to elucidate it by tracking melatonin receptor activity in male and female starlings housed in a semi-natural environment and permitted to breed. Males and females all exhibited high melatonin receptor activity in Area X during short days at the start of the breeding season, and maintained this high activity during photostimulation until females laid eggs. At this point the females down-regulated melatonin receptor activity in Area X, whereas the males maintained high activity until later on in the breeding season. Mel 1b was the most abundantly expressed of the 3 known melatonin receptor subtypes in Area X. There was a positive correlation between the expression of Mel 1b and the transcription factor ZENK, indicating that high melatonin receptor expression is correlated with high activity of Area X. Overall, we observed a gradual termination of activity in Area X as the breeding season progressed, but the timing of termination was different between the sexes.

Effects of Parental Experience and Age on Expression of Prolactin, Vasoactive Intestinal Peptide and their Receptors in a Biparental Bird (Columba livia).

As animals gain parental experience, they often show more rapid and efficient parental care responses that likely improve offspring survival and fitness. Changes in circulating hormones that underlie reproductive behaviors, including prolactin, have been found to correlate with parental experience in birds and mammals. Altered responsiveness to prolactin in key behavioral centers of the brain may also underlie the effects of experience on parental behaviors. Further, experience may also affect responsiveness to prolactin stimulatory hormones, such as hypothalamic vasoactive intestinal peptide (VIP). While experience has been shown to upregulate neural prolactin receptors and responsiveness in rodents, its effects on prolactin receptor gene expression remain unstudied in birds. To address this, we examined gene expression of pituitary prolactin, hypothalamic prolactin receptors in the preoptic area, hypothalamic VIP, and pituitary VIP receptors in both sexes of the biparental rock dove (Columba livia) when birds were not actively nesting. As age and parental experience are often confounded (i.e.,experienced parents tend to be older than their inexperienced counterparts), we measured gene expression in birds of varying combinations of age (0.6-3 years) and prior reproductive experience (0-12 chicks raised). We found that increasing experience with chicks correlated with lower PRLR expression in the preoptic area, and age correlated with lower VIP expression in birds of both sexes. Pituitary PRL and VIPR expression was not associated with parental experience or age. These results suggest there may be persistent effects of experience and age on neural responsiveness to, and regulation of, prolactin in birds.

Providing height to pullets does not influence hippocampal dendritic morphology or brain-derived neurotrophic factor at the end of the rearing period.

Pullets reared with diverse behavioral experiences are faster to learn spatial cognition tasks and acclimate more successfully to laying environments with elevated structures. However, the neural underpinnings of the improved spatial abilities are unclear. The objective of this study was to determine whether providing structural height in the rearing environment affected the development of the hippocampus and whether hippocampal neural metrics correlated with individual behavior on spatial cognition tasks. Female Dekalb White pullets were reared in a floor pen (FL), single-tiered aviary (ST), or two-tiered aviary (TT; 5 pens/treatment). Pullets completed floor-based Y-maze and elevated visual cliff tasks to evaluate depth perception at 15 and 16 wk, respectively. At 16 wk, brains were removed for Golgi-Cox staining (n = 12 for FL, 13 for ST, 13 total pullets for TT; 2 to 3 pullets/pen) and qPCR to measure gene expression of brain-derived neurotrophic factor (BDNF; n = 10 for FL, 11 for ST, and 9 pullets for TT). Rearing environment did not affect various morphometric outcomes of dendritic arborization, including Sholl profiles; mean dendritic length; sum dendritic length; number of dendrites, terminal tips, or nodes; soma size; or BDNF mRNA expression (P > 0.05). Hippocampal subregion did affect dendritic morphology, with multipolar neurons from the ventral subregion differing in several characteristics from multipolar neurons in the dorsomedial or dorsolateral subregions (P < 0.05). Neural metrics did not correlate with individual differences in behavior during the spatial cognition tasks. Overall, providing height during rearing did not affect dendritic morphology or BDNF at 16 wk of age, but other metrics in the hippocampus or other brain regions warrant further investigation. Additionally, other structural or social components or the role of animal personality are areas of future interest for how rearing environments influence pullet behavior.

Uncovering the Sex-Specific Endocrine Responses to Reproduction and Parental Care.

Hormones mediate physiological and behavioral changes in adults as they transition into reproduction. In this study, we characterize the circulating levels of five key hormones involved in reproduction in rock doves (Columba livia): corticosterone, progesterone, estradiol, testosterone, and prolactin using univariate and multivariate approaches. We show similar patterns as previous studies in the overall patterns in circulating levels of these hormones, i.e., testosterone (males) and estradiol (females) high during nest-building or egg-laying, prolactin increasing at mid-incubation and peaking at hatching (both sexes), and elevated corticosterone levels in later incubation and early nestling development. In our investigation of hormone co-variation, we find a strong correlation between prolactin and corticosterone across sampling stages and similarities in earlier (early to mid-incubation) compared to later (late incubation to nestling d9) sampling stages in males and females. Finally, we utilized experimental manipulations to simulate nest loss or altered caregiving lengths to test whether external cues, internal timing, or a combination of these factors contributed most to hormone variation. Following nest loss, we found that both males and females responded to the external cue. Males generally responded quickly following nest loss by increasing circulating testosterone, but this response was muted when nest loss occurred early in reproduction. Similar treatment type, e.g., removal of eggs, clustered similarly in hormone space. These results suggest internal drivers limited male response early in reproduction to nest loss. In contrast, circulating levels of these hormones in females either did not change or decreased following nest manipulation suggesting responsiveness to external drivers, but unlike males, this result suggests that reproductive processes were decreasing.

Isolating the Role of Corticosterone in the Hypothalamic-Pituitary-Gonadal Transcriptomic Stress Response.

Investigation of the negative impacts of stress on reproduction has largely centered around the effects of the adrenal steroid hormone, corticosterone (CORT), and its influence on a system of tissues vital for reproduction-the hypothalamus of the brain, the pituitary gland, and the gonads (the HPG axis). Research on the action of CORT on the HPG axis has predominated the stress and reproductive biology literature, potentially overshadowing other influential mediators. To gain a more complete understanding of how elevated CORT affects transcriptomic activity of the HPG axis, we experimentally examined its role in male and female rock doves (Columba livia). We exogenously administrated CORT to mimic circulating levels during the stress response, specifically 30 min of restraint stress, an experimental paradigm known to increase circulating CORT in vertebrates. We examined all changes in transcription within each level of the HPG axis as compared to both restraint-stressed birds and vehicle-injected controls. We also investigated the differential transcriptomic response to CORT and restraint-stress in each sex. We report causal and sex-specific effects of CORT on the HPG transcriptomic stress response. Restraint stress caused 1567 genes to uniquely differentially express while elevated circulating CORT was responsible for the differential expression of 304 genes. Only 108 genes in females and 8 in males differentially expressed in subjects that underwent restraint stress and those who were given exogenous CORT. In response to elevated CORT and restraint-stress, both sexes shared the differential expression of 5 genes, KCNJ5, CISH, PTGER3, CEBPD, and ZBTB16, all located in the pituitary. The known functions of these genes suggest potential influence of elevated CORT on immune function and prolactin synthesis. Gene expression unique to each sex indicated that elevated CORT affected more gene transcription in females than males (78 genes versus 3 genes, respectively). To our knowledge, this is the first study to isolate the role of CORT in HPG genomic transcription during a stress response. We present an extensive and openly accessible view of the role corticosterone in the HPG transcriptomic stress response. Because the HPG system is well conserved across vertebrates, these data have the potential to inspire new therapeutic strategies for reproductive dysregulation in multiple vertebrate systems, including our own.

Lab and field experiments: are they the same animal?

To advance our understanding of biological processes we often plan our experiments based on published data. This can be confusing though, as data from experiments performed in a laboratory environment are sometimes different from, or completely opposite to, findings from similar experiments performed in the "real world". In this mini-review, we discuss instances where results from laboratory experiments differ as a result of laboratory housing conditions, and where they differ from results gathered in the field environment. Experiments involving endocrinology and behavior appear to be particularly susceptible to influence from the environment in which they are performed. As such, we have attempted to promote discussion of the influence of housing environment on the reproductive axis, circadian biology and behavior, immune function, stress biology, neuroplasticity and photoperiodism. For example, why should a rodent species be diurnal in one housing environment yet nocturnal in another? Are data that are gathered from experiments in the laboratory applicable to the field environment, and vice-versa? We hope not only to highlight the need for experiments in both lab and field when looking at complex biological systems, but also to promote frank discussion of discordant data. Perhaps, just as study of individual variation has been gaining momentum in recent years, data from variation between experimental arenas can provide us with novel lines of research.

Environmentally relevant concentrations of bifenthrin affect the expression of estrogen and glucocorticoid receptors in brains of female western mosquitofish.

In recent decades, pyrethroid pesticides have been deemed a safer alternative to previously used pesticides. While some evidence supports this assumption in mammals and birds, exposure to certain pyrethroids can affect concentrations of hormones vital to reproduction in fish. Thus, we hypothesized that pyrethroid exposure impacts fish reproductive behavior and the expression of genes associated with reproduction. We tested our hypothesis by examining effects of the widely used pyrethroid pesticide, bifenthrin, on the reproductive behaviors of the broadly distributed livebearing western mosquitofish, Gambusia affinis. We exposed sexually mature female fish to one of five environmentally relevant concentrations of bifenthrin and conducted behavioral assays to assess reproductive, social, and space use behaviors before and after exposure. We did not detect changes in behaviors measured in response to bifenthrin. However, exposure was associated with increased expression of an estrogen receptor gene (ER-α) and glucocorticoid receptor (GR) in brain tissue at bifenthrin concentrations at concentrations of 5.90 and 24.82 ng/L, and 5.90 and 12.21 ng/L, respectively. Our study supports the perspective that the use of multiple endpoints through integrative approaches is essential for understanding the cumulative impact of pollutants. Integrating physiological, morphological, and behavioral investigations of nonlethal concentrations of pollutants like bifenthrin may heighten our potential to predict their impact on individuals, populations, and communities.

Seasonal differences in hypothalamic EGR-1 and GnIH expression following capture-handling stress in house sparrows (Passer domesticus).

Stress is a known inhibitor of reproductive function. The mechanisms by which stress acts to influence the reproductive axis have been intensely studied and appear to be extremely varied. Gonadotropin-releasing hormone (GnRH) is a critical component of the vertebrate reproductive axis and directly causes pituitary gonadotropin synthesis and release. A second neuropeptide, gonadotropin-inhibitory hormone (GnIH), directly inhibits pituitary gonadotropin synthesis and release in birds. We hypothesized that stress effects upon reproduction are mediated via the hypothalamic GnIH system. We examined the effects of capture-handling stress in the hypothalamus of male and female adult house sparrows (Passer domesticus) at the start (spring) and end of the breeding season (fall). We quantified numbers of GnIH neurons to provide an estimate of hypothalamic GnIH content. In addition, we quantified the expression of the protein product of the immediate-early gene, EGR-1, using this as an indicator of neuronal activation. We saw an increase in EGR-1 positive cells in the paraventricular nuclei of stressed birds as opposed to controls at both collecting times, but this stress response was more apparent in the spring as opposed to the fall. There were more GnIH-positive neurons in fall birds versus those sampled in the spring, and a significant increase in GnIH positive neurons was seen in stressed birds only in spring. GnIH cells show little to no activation of EGR-1, suggesting that EGR-1 is not involved in GnIH transcription in response to capture-handling stress. These data imply an influence of stress upon the paraventricular nucleus and the GnIH system that changes over the annual cycle of reproduction.

Nonapeptide Receptor Distributions in Promising Avian Models for the Neuroecology of Flocking.

Collective behaviors, including flocking and group vocalizing, are readily observable across a diversity of free-living avian populations, yet we know little about how neural and ecological factors interactively regulate these behaviors. Because of their involvement in mediating a variety of social behaviors, including avian flocking, nonapeptides are likely mediators of collective behaviors. To advance the neuroecological study of collective behaviors in birds, we sought to map the neuroanatomical distributions of nonapeptide receptors in three promising avian models that are found across a diversity of environments and widely ranging ecological conditions: European starlings, house sparrows, and rock doves. We performed receptor autoradiography using the commercially available nonapeptide receptor radioligands, 125I-ornithine vasotocin analog and 125I-linear vasopressin antagonist, on brain tissue sections from wild-caught individuals from each species. Because there is known pharmacological cross-reactivity between nonapeptide receptor subtypes, we also performed a novel, competitive-binding experiment to examine the composition of receptor populations. We detected binding in numerous regions throughout the brains of each species, with several similarities and differences worth noting. Specifically, we report that all three species exhibit binding in the lateral septum, a key brain area known to regulate avian flocking. In addition, sparrows and starlings show dense binding in the dorsal arcopallium, an area that has received scant attention in the study of social grouping. Furthermore, our competitive binding results suggest that receptor populations in sparrows and starlings differ in the lateral septum versus the dorsal arcopallium. By providing the first comprehensive maps of nonapeptide receptors in European starlings, house sparrows, and rock doves, our work supports the future use of these species as avian models for neuroecological studies of collective behaviors in wild birds.

Widespread patterns of sexually dimorphic gene expression in an avian hypothalamic-pituitary-gonadal (HPG) axis.

The hypothalamic-pituitary-gonadal (HPG) axis is a key biological system required for reproduction and associated sexual behaviors to occur. In the avian reproductive model of the rock dove (Columba livia), we characterized the transcript community of each tissue of the HPG axis in both sexes, thereby significantly expanding our mechanistic insight into HPG activity. We report greater sex-biased differential expression in the pituitary as compared to the hypothalamus, with multiple genes more highly expressed in the male pituitary being related to secretory function, and multiple genes more highly expressed in the female pituitary being related to reproduction, growth, and development. We report tissue-specific and sex-biased expression in genes commonly investigated when studying reproduction, highlighting the need for sex parity in future studies. In addition, we uncover new targets of investigation in both sexes, which could potentially change our understanding of HPG function.

Neural Versus Gonadal GnIH: Are they Independent Systems? A Mini-Review.

Based on research in protochordates and basal vertebrates, we know that communication across the first endocrine axes likely relied on diffusion. Because diffusion is relatively slow, rapid responses to some cues, including stress-related cues, may have required further local control of axis outputs (e.g., steroid hormone production by the gonads). Despite the evolution of much more efficient circulatory systems and complex nervous systems in vertebrates, production of many "neuro"transmitters has been identified outside of the hypothalamus across the vertebrate phylogeny and these neurotransmitters are known to locally regulate endocrine function. Our understanding of tissue-specific neuropeptide expression and their role coordinating physiological/behavioral responses of the whole organism remains limited, in part, due to nomenclature and historic dogma that ignores local regulation of axis output. Here, we review regulation of gonadotropin-inhibitory hormone (GnIH) across the reproductive axis in birds and mammals to bring further attention to context-dependent disparities and similarities in neuropeptide production by the brain and gonads. We find that GnIH responsiveness to cues of stress appears conserved across species, but that the response of specific tissues and the direction of GnIH regulation varies. The implications of differential regulation across tissues remain unclear in most studies, but further work that manipulates and contrasts function in different tissues has the potential to inform us about both organism-specific function and endocrine axis evolution.