Central prolactin receptor distribution and pSTAT5 activation patterns in breeding and non-breeding zebra finches (Taeniopygia guttata).

The hormone prolactin has many diverse functions across taxa such as osmoregulation, metabolism, and reproductive behavior. In ring doves, central prolactin action is important for parental care and feeding behavior. However, there is a considerable lack of information on the distribution of the prolactin receptor (PRLR) in the avian CNS to test the hypothesis that prolactin mediates these and other functions in other birds. In order to advance this research, we collected brains from breeding and non-breeding zebra finches to map the PRLR distribution using immunohistochemistry. We found PRLRs are distributed widely across the brain, both in hypothalamic sites known to regulate parental care and feeding, but also in many non-hypothalamic sites, including the tectofugal visual pathway, song system regions, reward associated areas, and pallium. This raises the possibility that prolactin has other functions throughout the brain that are not necessarily related to feeding or parental care. In addition, we also stained brains for pSTAT5, a transcription factor which is expressed when the PRLR is activated and is used as a marker for PRLR activity. We found several notable differences in pSTAT5 activity due to the breeding state of the animal, in both directions, further supporting the hypothesis that prolactin has many diverse functions in the brain both within and outside times of breeding. Together, this study represents the first essential step to inform the design of causative studies which manipulate PRLR-expressing cells to test their role in a wide variety of behaviors and other physiological functions.

Sexual and pairing partner preference in birds and other animals.

Research on hormonal involvement in animals' preferences for mating or pairing with same- or other-sex partners has been among the topics featured in Hormones and Behavior over the years. In several species of non-pair-forming mammals, there is good evidence that the early sex steroid environment has an organizational effect on later sexual partner preference. Research with zebra finches, a pair-forming species, shows a likely early estrogenic organizational hormone effect on pairing partner preference, an effect that can also interact with the early social environment to determine adult pairing preference. Experiments with two amphibian and fish species suggest that activational hormone effects (effects of the hormone milieu in adulthood) may regulate sexual partner preference. As a complement to the research on hormonal involvement, a growing body of theoretical and empirical research addresses the possible adaptive functions of the same-sex sexual and pairing behavior observed in many wild animals. Such advances have important implications for conceptualizing neuroendocrine mechanisms of partner preferences.

Do nonapeptides regulate parental care depending on experience in zebra finches?

Recent research suggests that the nonapeptide neurohormones regulate parental behaviors in a diverse array of vertebrates. However, it remains unclear how these neurohormones regulate parental care among birds, especially those which exhibit biparental care, or whether hormonal effects are contingent on a bird's previous experience as a parent. We measured the effects of nonapeptides on parental behaviors by peripherally injecting, over three treatment days, a short-acting nonapeptide receptor antagonist (OTA) or a saline control into breeding pairs of zebra finches (Taeniopygia guttata) that either did or did not have previous parental experience. We then compared how the duration of parental behaviors changed over the five days of observation (including one day before and two days after injections were administered). To compare treatment effects on parental outcomes, we also measured chick growth and mortality rates for each pair. There was a nearly significant interaction between treatment and experience for the amount of time birds spent in the nest, with time in the nest declining across the experiment inexperienced and experienced OTA birds. There was also a significant treatment by trial day interaction for nest guarding and a treatment by experience by trial day interaction for nest maintenance. Chicks reared by parents that received the OTA had significantly lower growth rates than chicks reared by control parents and, among experienced birds, higher mortality relative to control birds. Together, these results provide some support for the hypothesis that nonapeptides play a role in regulating parental outcomes and some parental behaviors in both experienced and inexperienced zebra finches.

Co-opting evo-devo concepts for new insights into mechanisms of behavioural diversity.

We propose that insights from the field of evolutionary developmental biology (or 'evo-devo') provide a framework for an integrated understanding of the origins of behavioural diversity and its underlying mechanisms. Towards that goal, in this Commentary, we frame key questions in behavioural evolution in terms of molecular, cellular and network-level properties with a focus on the nervous system. In this way, we highlight how mechanistic properties central to evo-devo analyses - such as weak linkage, versatility, exploratory mechanisms, criticality, degeneracy, redundancy and modularity - affect neural circuit function and hence the range of behavioural variation that can be filtered by selection. We outline why comparative studies of molecular and neural systems throughout ontogeny will provide novel insights into diversity in neural circuits and behaviour.

Lowering prolactin reduces post-hatch parental care in male and female zebra finches (Taeniopygia guttata).

Parental care is a widespread phenomenon observed in many diverse taxa. Neuroendocrine systems have long been thought to play an important role in stimulating the onset of parental behavior. In most birds with altricial young, circulating prolactin (PRL) levels are low during non-breeding times and significantly increase during late incubation and early post-hatch chick care. Because of this pattern, PRL has been suggested to be involved in the initiation of parental care in birds, but rarely has this hypothesis been causally tested. To begin testing the hypothesis, we inhibited the release of endogenous PRL with bromocriptine (BR) on the 3days prior to hatching in incubating parents and the first 2days of post-hatch care, when PRL was found to be highest in zebra finches. Nest temperatures were recorded during all 5days and parental behavior was recorded on days 1-2 post-hatch. In addition to hormonal systems, reproductive experience may also influence parental care; therefore, we tested age-matched inexperienced and experienced pairs in each group. BR either eliminated or drastically reduced chick brooding and feeding behavior, resulting in decreased nest temperatures on days 1 and 2 post-hatch. Experienced control birds fed chicks more than inexperienced birds and control females fed more than males. Chick feeding behavior was positively correlated in control male-female pairs, but not in BR pairs. This is one of the few causal studies to demonstrate that PRL is necessary for post-hatch care in a biparental songbird, and is the first to show this effect in zebra finches.

Early life manipulations of vasopressin-family peptides alter vocal learning.

Vocal learning from social partners is crucial for the successful development of communication in a wide range of species. Social interactions organize attention and enhance motivation to learn species-typical behaviour. However, the neurobiological mechanisms connecting social motivation and vocal learning are unknown. Using zebra finches (Taeniopygia guttata), a ubiquitous model for vocal learning, we show that manipulations of nonapeptide hormones in the vasopressin family (arginine vasotocin, AVT) early in development can promote or disrupt both song and social motivation. Young male zebra finches, like human infants, are socially gregarious and require interactive feedback from adult tutors to learn mature vocal forms. To investigate the role of social motivational mechanisms in song learning, in two studies, we injected hatchling males with AVT or Manning compound (MC, a nonapeptide receptor antagonist) on days 2-8 post-hatching and recorded song at maturity. In both studies, MC males produced a worse match to tutor song than controls. In study 2, which experimentally controlled for tutor and genetic factors, AVT males also learned song significantly better compared with controls. Furthermore, song similarity correlated with several measures of social motivation throughout development. These findings provide the first evidence that nonapeptides are critical to the development of vocal learning.

Developmental effects of vasotocin and nonapeptide receptors on early social attachment and affiliative behavior in the zebra finch.

Zebra finches demonstrate selective affiliation between juvenile offspring and parents, which, like affiliation between pair partners, is characterized by proximity, vocal communication and contact behaviors. This experiment tested the hypothesis that the nonapeptide arginine vasotocin (AVT, avian homologue of vasopressin) and nonapeptide receptors play a role prior to fledging in the development of affiliative behavior. Zebra finch hatchlings of both sexes received daily intracranial injections (post-hatch days 2-8) of either AVT, Manning Compound (MC, a potent V1a receptor antagonist) or saline (vehicle control). The social development of both sexes was assessed by measuring responsiveness to isolation from the family and subsequent reunion with the male parent after fledging. In addition, we assessed the changes in affiliation with the parents, unfamiliar males, and unfamiliar females each week throughout juvenile development. Compared to controls, MC subjects showed decreased attachment to the parents and MC males did not show the normal increase in affiliative interest in opposite sex individuals as they reached reproductive maturity. In contrast, AVT subjects showed a sustained affiliative interest in parents throughout development, and males showed increased interest in opposite sex conspecifics as they matured. These results provide the first evidence suggesting that AVT and nonapeptide receptors play organizational roles in social development in a bird.

Relationship between prolactin, reproductive experience, and parental care in a biparental songbird, the zebra finch (Taeniopygia guttata).

Hormonal systems have long been thought to play an important role in stimulating the onset of parental behavior, a critical component of reproductive success in a variety of taxa. Elevations in the peptide hormone prolactin (PRL) have been repeatedly positively correlated with the onset and maintenance of parental care across vertebrate species. A causal role for PRL in parental care has been established in several mammalian species, but less evidence for a causal role of PRL and parental care exists in birds. The zebra finch, a socially monogamous, biparental songbird, is an exceptionally useful animal model to study parental care and other close social relationships. Both sexes share parental care equally, exhibit the same parental behaviors, and show a marked improvement in breeding success with experience. We hypothesize that PRL is critically involved in the expression of zebra finch parental care and predict that circulating PRL levels will increase with breeding experience. To begin testing this, we measured plasma PRL concentrations in 14 male-female zebra finch pairs (N=28) across two breeding cycles, using a repeated measures design. PRL was measured in the birds' first, reproductively inexperienced, breeding cycle beginning at courtship and extending through chick fledging. PRL was measured again during the birds' second, reproductively experienced, breeding cycle, beginning with egg laying until chick fledging. We found that plasma PRL is significantly elevated from non-breeding concentrations during late incubation and early post-hatch care and that this elevation is greater in the reproductively experienced cycle compared to the inexperienced cycle. Findings of this study will be used to inform hypotheses and predictions for future experimental manipulations of PRL during parental care.

Prolactin is related to individual differences in parental behavior and reproductive success in a biparental passerine, the zebra finch (Taeniopygia guttata).

Variation in parental care can lead to important fitness consequences. The endocrine system is known to regulate physiological and behavioral reproductive traits that are important contributors to lifetime reproductive success. However, the hormonal basis of variation in avian parental care is still not well understood. Plasma prolactin (PRL) concentrations are generally high during post-hatch parental care in birds, and may be a candidate mechanism that regulates variation in parental care and other reproductive success outcomes. Here we analyze the relationship between PRL, parental behavior (chick brooding and feeding) and reproductive success outcomes (clutch size, number of chicks hatched, and chick survival) for the first time in the zebra finch (Taeniopygia guttata). Birds were given cabergoline, a dopamine agonist traditionally used to lower prolactin in mammals, or vehicle in their food. Cabergoline had no effect on prolactin concentrations, but across both groups we found that PRL is positively correlated with parental behavior, number of chicks hatched, and chick survival, but not clutch size. Results from this study will inform hypotheses and predictions for future manipulation studies which test for a causal role for PRL in parental traits.

Sexual dimorphism in body size and the origin of sex-determination systems.

Diverse sex-determining systems occur in vertebrates, including environmental sex determination (ESD), genetic sex determination (GSD) of type XX/XY (heterogametic males), and GSD of type ZZ/ZW (heterogametic females). The origins of the two genetic types are poorly understood. We use protected invasion theory to derive a model that generates testable predictions about the origins of the two genetic types from ESD. Protected invasion theory predicts biases in the evolutionary origins of new traits by focusing on the probability that a selectively favored trait will avoid loss by genetic drift when rare. We show that the theory makes predictions about the conditions under which XY or ZW systems are more likely to arise from an ancestral state of ESD. In particular, assuming that there is an average trend toward increasing body size in lineages, the origins of XY systems are predicted to be accompanied by increases in male∶female body size ratio. In contrast, ZW systems are predicted to be accompanied by decreases in male∶female body size ratio. We find support for these predictions in the form of a marked association among vertebrates between sex-determining system and body size dimorphism in paired comparisons independent of shared phylogeny.