Varied impacts of social relationships on neuroendocrine state.

Social relationships, affiliative social attachments, are important for many species. The best studied types of relationships are monogamous pair bonds. However, it remains unclear how generalizable models of pair bonding are across types of social attachments. Zebra finches are a fascinating system to explore the neurobiology of social relationships because they form various adult bonds with both same- and opposite-sex partners. To test whether different bonds are supported by a single brain network, we quantified individuals' neuroendocrine state after either 24 h or 2 weeks of co-housing with a novel same- or opposite-sex partner. We defined neuroendocrine state by the expression of 22 genes related to 4 major signaling pathways (dopamine, steroid, nonapeptide, and opioid) in six brain regions associated with affiliation or communication [nucleus accumbens (NAc), nucleus taeniae of the amygdala (TnA), medial preoptic area (POM), and periaqueductal gray (PAG), ventral tegmental area, and auditory cortex]. Overall, we found dissociable effects of social contexts (same- or opposite-sex partnerships) and duration of co-housing. Social bonding impacted the neuroendocrine state of four regions in males (NAc, TnA, POM, and PAG) and three regions in females (NAc, TnA, and POM). Monogamous pair bonding specifically appeared to impact male NAc. However, the patterns of gene expression in zebra finches were different than has previously been reported in mammals. Together, our results support the view that there are numerous mechanisms regulating social relationships and highlight the need to further our understanding of how social interactions shape social bonds.

The singing question: re-conceptualizing birdsong.

Birdsong has been the subject of broad research from a variety of sub-disciplines and has taught us much about the evolution, function, and mechanisms driving animal communication and cognition. Typically, birdsong refers to the specialized vocalizations produced by oscines. Historically, much of the research on birdsong was conducted in north temperate regions (specifically in Europe and North America) leading to multiple biases. Due to these historic biases these vocalizations are generally considered to be highly sexually dimorphic, heavily shaped by sexual selection and essential for courtship and territoriality. Song is also typically defined as a learned trait shaped by cultural evolution. Together, this framework focuses research specifically on males, particularly during the north temperate breeding season - reflecting and thereby reinforcing this framework. The physiological underpinnings of song often emphasize the role of the hypothalamic-pituitary-gonadal axis (associated with breeding changes) and the song control system (underlying vocal learning). Over the years there has been great debate over which features of song are essential to the definition of birdsong, which features apply broadly to contexts outside males in the north temperate region, and over the importance of having a definition at all. Importantly, the definitions we use can both guide and limit the progress of research. Here, we describe the history of these definitions, and how these definitions have directed and restricted research to focus on male song in sexually selected contexts. Additionally, we highlight the gaps in our scientific knowledge, especially with respect to the function and physiological mechanisms underlying song in females and in winter, as well as in non-seasonally breeding species. Furthermore, we highlight the problems with using complexity and learning as dichotomous variables to categorize songs and calls. Across species, no one characteristic of song - sexual dimorphism, seasonality, complexity, sexual selection, learning - consistently delineates song from other songbird vocal communication. We provide recommendations for next steps to build an inclusive information framework that will allow researchers to explore nuances in animal communication and promote comparative research. Specifically, we recommend that researchers should operationalize the axis of variation most relevant to their study/species by identifying their specific question and the variable(s) of focus (e.g. seasonality). Researchers should also identify the axis (axes) of variation (e.g. degree of control by testosterone) most relevant to their study and use language consistent with the question and axis (axes) of variation (e.g. control by testosterone in the seasonal vocal production of birds).

Reciprocal processes of sensory perception and social bonding: an integrated social-sensory framework of social behavior.

Organisms filter the complexity of natural stimuli through their individual sensory and perceptual systems. Such perceptual filtering is particularly important for social stimuli. A shared "social umwelt" allows individuals to respond appropriately to the expected diversity of cues and signals during social interactions. In this way, the behavioral and neurobiological mechanisms of sociality and social bonding cannot be disentangled from perceptual mechanisms and sensory processing. While a degree of embeddedness between social and sensory processes is clear, our dominant theoretical frameworks favor treating the social and sensory processes as distinct. An integrated social-sensory framework has the potential to greatly expand our understanding of the mechanisms underlying individual variation in social bonding and sociality more broadly. Here we leverage what is known about sensory processing and pair bonding in two common study systems with significant species differences in their umwelt (rodent chemosensation and avian acoustic communication). We primarily highlight that (1) communication is essential for pair bond formation and maintenance, (2) the neural circuits underlying perception, communication and social bonding are integrated, and (3) candidate neuromodulatory mechanisms that regulate pair bonding also impact communication and perception. Finally, we propose approaches and frameworks that more fully integrate sensory processing, communication, and social bonding across levels of analysis: behavioral, neurobiological, and genomic. This perspective raises two key questions: (1) how is social bonding shaped by differences in sensory processing?, and (2) to what extent is sensory processing and the saliency of signals shaped by social interactions and emerging relationships?

Discrimination of natural acoustic variation in vocal signals.

Studies of acoustic communication often focus on the categories and units of vocalizations, but subtle variation also occurs in how these signals are uttered. In human speech, it is not only phonemes and words that carry information but also the timbre, intonation, and stress of how speech sounds are delivered (often referred to as "paralinguistic content"). In non-human animals, variation across utterances of vocal signals also carries behaviorally relevant information across taxa. However, the discriminability of these cues has been rarely tested in a psychophysical paradigm. Here, we focus on acoustic communication in the zebra finch (Taeniopygia guttata), a songbird species in which the male produces a single stereotyped motif repeatedly in song bouts. These motif renditions, like the song repetitions of many birds, sound very similar to the casual human listener. In this study, we show that zebra finches can easily discriminate between the renditions, even at the level of single song syllables, much as humans can discriminate renditions of speech sounds. These results support the notion that sensitivity to fine acoustic details may be a primary channel of information in zebra finch song, as well as a shared, foundational property of vocal communication systems across species.

What's in a Moment: What Can Be Learned About Pair Bonding From Studying Moment-To-Moment Behavioral Synchrony Between Partners?

Our understanding of the behavioral and physiological mechanisms of monogamy largely comes from studies of behavioral interactions unique to pair-bonded individuals. By focusing on these highly marked behaviors, a remarkable conservation in the mechanisms underlying pair bonding has been revealed; however, we continue to know very little about the range of behavioral and neurobiological mechanisms that could explain the great diversity of pair-bonding phenotypes that exists both within and across species. In order to capture the dynamic nature of bonds over time and across contexts, we need specific, operationally-defined behavioral variables relevant across such a diversity of scenarios. Additionally, we need to be able to situate these behavioral variables within broader frameworks that allow us to interpret and compare patterns seen across species. Here I review what is known about behavioral synchrony with respect to pair bonding and discuss using synchrony as such a variable as well as a framework to expand on our understanding of pair bonding across timescales, contexts and species. First, I discuss the importance of behavioral synchrony and parental coordination for reproductive success in monogamous biparental bird species. Second, I highlight research documenting the critical importance of interpersonal coordination for human social relationships. Finally, I present recent work that experimentally bridges these lines of research by quantifying moment-to-moment behavioral synchrony during brief social interactions in zebra finch dyads. All together, these distinct perspectives support the notion that synchrony (1) is a shared premise for sociality across species, (2) is deeply shaped by social experiences, and (3) exists across timescales, behaviors, and levels of physiology. Conceptualizing pair bonding through the framework of behavioral synchrony is likely to facilitate a deeper understanding of the nuances of how social experiences and interactions impact the brain and behavior.

Monogamy in a Moment: How do Brief Social Interactions Change Over Time in Pair-Bonded Zebra Finches (Taeniopygia guttata)?

Research on monogamy has largely focused on marked behaviors that are unique to pair bonded partners. However, these marked behaviors represent only a subset of the pair-directed behaviors that partners engage in; the influence of pair bonding on mundane or subtle social interactions among partners remains largely unknown. In this study, we describe the changes that occur during brief social reunions (or greets) over the course of pair bonding in zebra finches. We quantified pair-directed behavior during 5-min reunions from three stages of pair bonding: initial pairing (between 4 and 72 h), early pairing (1-2 weeks), and late pairing (>1 month). These social interactions were operationalized in multiple ways. First, we quantified the overall activity levels (call and movement rates) for both the male and female. Overall, females were more active than males, but for both males and females calling activity was highest at initial pairing. We quantified behavioral coordination between partners in two ways: (1) similarity in call and movement rates between partners and (2) temporal synchrony of calls and movements between partners (via sliding correlation coefficients of time-stamped calls and movements). Overall, there were no effects of pairing stage on behavioral coordination. Finally, we used principal component analyses to disentangle behavioral coordination from the activity levels of the male and female. These results contribute to a growing line of evidence that male and female zebra finches differentially contribute to social dynamics and highlight the influence of pair bonding on the development of social dynamics. Furthermore, our preliminary analyses raise the hypothesis that behavioral coordination during the earliest phases of pairing is modulated by the extent and nature of prior experience. Overall, while behavioral coordination is clearly important for many salient interactions such as duetting, courtship displays, and biparental care, the significance of mundane social interactions for monogamous partnerships remains largely unknown.

Familiarity enhances moment-to-moment behavioral coordination in zebra finch (Taeniopygia guttata) dyads.

An individual's ability to respond to and align with the behavior of others is a fundamental component of social behavior. Zebra finches form lifelong monogamous pair bonds; however, zebra finches are also gregarious and can form strong social bonds with same-sex conspecifics. Here, we quantified behavior during brief 10-min reunions for males and females in five types of social conditions: monogamously bonded opposite-sex partners, familiar same-sex, familiar opposite-sex, novel same-sex, and novel opposite-sex dyads. We analyzed these interactions in three ways. First, we quantified overall activity levels (call and movement rates) for each individual. Second, we measured how coordinated calls and movements were by calculating (a) the percent difference in activity rates as an estimate of how similar calling and movement activity were between individuals within a dyad, and (b) the sliding correlation coefficients for time-stamped calls and movements for each dyad. Finally, we described multimodal behavioral profiles of coordination using principal component analyses. Overall, females were more active than males. For both females and males, activity levels as well as the coordination of calls and movements were significantly affected by social condition. In general, monogamous partners, female familiar same-sex dyads and familiar opposite-sex dyads were the most coordinated. This effect of familiarity shows that moment-to-moment behavioral coordination can be influenced by prior social experiences. Quantifying patterns of coordination or social synchrony may prove valuable for understanding the effects of social experience on brain and behavior. (PsycInfo Database Record (c) 2020 APA, all rights reserved).

Water restriction influences intra-pair vocal behavior and the acoustic structure of vocalisations in the opportunistically breeding zebra finch (Taeniopygia guttata).

Seasonally-breeding species experience significant and predictable shifts in vocal behaviour; however, it is unclear to what extent this is true for species that breed opportunistically. The Australian zebra finch is an opportunistically breeding species, which means individuals must time breeding bouts based on many environmental factors. Here we tested the effect of experimental water restriction, which suppresses reproductive readiness in zebra finches, on vocal behaviour of males and females. More specifically, we quantified the effect of water restriction on three parameters of vocal behaviour in pair-bonded zebra finches: vocal activity, patterns of vocal exchanges, and the acoustic structure of vocalisations (calls and male song). We found that water restriction caused a decrease in vocal output (both song and call rate). Additionally, water restriction affected the composition of male songs. However, there was no effect of water restriction on the patterns of calling exchanges for monogamous partners (temporal coordination and turn taking). Finally, water restriction had vocalisation- and sex-specific effects on the acoustic structure of song syllables and calls. Because the direction of these effects were vocalisation- and sex- specific, there may be different mechanisms underlying the effects of water restriction on acoustic structure depending on context. These results contribute to the growing body of research highlighting the rich communicative potential of bird calls. Our current results raise the hypothesis that zebra finches may use changes in vocal behaviour and/or the structure of vocalisations of their conspecifics when making breeding decisions.

Neuropeptide Y and orexin immunoreactivity in the sparrow brain coincide with seasonal changes in energy balance and steroids.

The transition between the breeding and nonbreeding states is often marked by a shift in energy balance. Despite this well-known shift in energy balance, little work has explored seasonal differences in the orexigenic neuropeptides that regulate food intake in wild animals. Here we tested the hypothesis that free-living male song sparrows (Melospiza melodia) show seasonal changes in energetic state, circulating steroids, and both neuropeptide Y (NPY) and orexin (OX) immunoreactivity. Nonbreeding song sparrows had more fat and muscle, as well as a ketone and triglyceride profile suggesting a greater reliance on lipid reserves. Breeding birds had higher plasma androgens; however, nonbreeding birds did maintain androgen precursors in circulation. Nonbreeding birds had more NPY immunoreactivity, specifically in three brain regions: lateral septum, bed nucleus of the stria terminalis, and ventral tegmental area. Furthermore, nonbreeding birds had more OX immunoreactivity in multiple brain regions. Taken together, the data indicate that a natural shift in energy balance is associated with changes in NPY and OX in a region-specific manner.

Rapid effects of sex steroids on zebra finch (Taeniopygia guttata) pair maintenance.

Although steroids are widely known to affect behavior through activation of nuclear/cytosolic receptors ("genomic" effects), steroids can also rapidly affect behavior via modulation of signal transduction pathways ("nongenomic," fast actions, or rapid effects). In zebra finches, there is evidence that sex steroids have context-specific effects on pair-maintenance behavior, on both acute and chronic timescales. Here, we quantified the effects of orally administered testosterone and 17ß-estradiol (E2) on pair-maintenance behavior. We show that E2 rapidly affects female, but not male, affiliative behavior profiles during a partner separation and reunion paradigm. More specifically, E2 rapidly (within 5-15 min of administration) increased females' spatial proximity to a partner. This effect was present regardless of breeding condition (water restriction or water ad libitum). Combined, these results contribute to a growing body of evidence implicating sex steroids in the regulation of prosocial behavior. (PsycINFO Database Record (c) 2018 APA, all rights reserved).

Relative salience of syllable structure and syllable order in zebra finch song.

There is a rich history of behavioral and neurobiological research focused on the 'syntax' of birdsong as a model for human language and complex auditory perception. Zebra finches are one of the most widely studied songbird species in this area of investigation. As they produce song syllables in a fixed sequence, it is reasonable to assume that adult zebra finches are also sensitive to the order of syllables within their song; however, results from electrophysiological and behavioral studies provide somewhat mixed evidence on exactly how sensitive zebra finches are to syllable order as compared, say, to syllable structure. Here, we investigate how well adult zebra finches can discriminate changes in syllable order relative to changes in syllable structure in their natural song motifs. In addition, we identify a possible role for experience in enhancing sensitivity to syllable order. We found that both male and female adult zebra finches are surprisingly poor at discriminating changes to the order of syllables within their species-specific song motifs, but are extraordinarily good at discriminating changes to syllable structure (i.e., reversals) in specific syllables. Direct experience or familiarity with a song, either using the bird's own song (BOS) or the song of a flock mate as the test stimulus, improved both male and female zebra finches' sensitivity to syllable order. However, even with experience, birds remained much more sensitive to structural changes in syllables. These results help to clarify some of the ambiguities from the literature on the discriminability of changes in syllable order in zebra finches, provide potential insight on the ethological significance of zebra finch song features, and suggest new avenues of investigation in using zebra finches as animal models for sequential sound processing.

Acoustic fine structure may encode biologically relevant information for zebra finches.

The ability to discriminate changes in the fine structure of complex sounds is well developed in birds. However, the precise limit of this discrimination ability and how it is used in the context of natural communication remains unclear. Here we describe natural variability in acoustic fine structure of male and female zebra finch calls. Results from psychoacoustic experiments demonstrate that zebra finches are able to discriminate extremely small differences in fine structure, which are on the order of the variation in acoustic fine structure that is present in their vocal signals. Results from signal analysis methods also suggest that acoustic fine structure may carry information that distinguishes between biologically relevant categories including sex, call type and individual identity. Combined, our results are consistent with the hypothesis that zebra finches can encode biologically relevant information within the fine structure of their calls. This study provides a foundation for our understanding of how acoustic fine structure may be involved in animal communication.

Do we hear what birds hear in birdsong?

Peter Marler's fascination with richness of birdsong included the notion that birds attended to some acoustic features of birdsong, likely in the time domain, which were inaccessible to human listeners. While a considerable amount is known about hearing and vocal communication in birds, how exactly birds perceive their auditory world still remains somewhat of a mystery. For sure, field and laboratory studies suggest that birds hear the spectral, gross temporal features (i.e. envelope) and perhaps syntax of birdsong much like we do. However, there is also ample anecdotal evidence that birds are consistently more sensitive than humans to at least some aspects of their song. Here we review several psychophysical studies supporting Marler's intuitions that birds have both an exquisite sensitivity to temporal fine structure and may be able to focus their auditory attention on critical acoustic details of their vocalizations. Zebra finches, Taeniopygia guttata, particularly, seem to be extremely sensitive to temporal fine structure in both synthetic stimuli and natural vocalizations. This finding, together with recent research highlighting the complexity of zebra finch vocalizations across contexts, raises interesting questions about what information zebra finches may be communicating in temporal fine structure. Together these findings show there is an acoustic richness in bird vocalizations that is available to birds but likely out of reach for human listeners. Depending on the universality of these findings, it raises questions about how we approach the study of birdsong and whether potentially significant information is routinely being encoded in the temporal fine structure of avian vocal signals.

Sex steroid profiles in zebra finches: Effects of reproductive state and domestication.

The zebra finch is a common model organism in neuroscience, endocrinology, and ethology. Zebra finches are generally considered opportunistic breeders, but the extent of their opportunism depends on the predictability of their habitat. This plasticity in the timing of breeding raises the question of how domestication, a process that increases environmental predictability, has affected their reproductive physiology. Here, we compared circulating steroid levels in various "strains" of zebra finches. In Study 1, using radioimmunoassay, we examined circulating testosterone levels in several strains of zebra finches (males and females). Subjects were wild or captive (Captive Wild-Caught, Wild-Derived, or Domesticated). In Study 2, using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we examined circulating sex steroid profiles in wild and domesticated zebra finches (males and females). In Study 1, circulating testosterone levels in males differed across strains. In Study 2, six steroids were detectable in plasma from wild zebra finches (pregnenolone, progesterone, dehydroepiandrosterone (DHEA), testosterone, androsterone, and 5α-dihydrotestosterone (5α-DHT)). Only pregnenolone and progesterone levels changed across reproductive states in wild finches. Compared to wild zebra finches, domesticated zebra finches had elevated levels of circulating pregnenolone, progesterone, DHEA, testosterone, androstenedione, and androsterone. These data suggest that domestication has profoundly altered the endocrinology of this common model organism. These results have implications for interpreting studies of domesticated zebra finches, as well as studies of other domesticated species.

Sex steroid profiles and pair-maintenance behavior of captive wild-caught zebra finches (Taeniopygia guttata).

Here, we studied the life-long monogamous zebra finch, to examine the relationship between circulating sex steroid profiles and pair-maintenance behavior in pairs of wild-caught zebra finches (paired in the laboratory for >1 month). We used liquid chromatography-tandem mass spectrometry to examine a total of eight androgens and progestins [pregnenolone, progesterone, dehydroepiandrosterone (DHEA), androstenediol, pregnan-3,17-diol-20-one, androsterone, androstanediol, and testosterone]. In the plasma, only pregnenolone, progesterone, DHEA, and testosterone were above the limit of quantification. Sex steroid profiles were similar between males and females, with only circulating progesterone levels significantly different between the sexes (female > male). Circulating pregnenolone levels were high in both sexes, suggesting that pregnenolone might serve as a circulating prohormone for local steroid synthesis in zebra finches. Furthermore, circulating testosterone levels were extremely low in both sexes. Additionally, we found no correlations between circulating steroid levels and pair-maintenance behavior. Taken together, our data raise several interesting questions about the neuroendocrinology of zebra finches.

Neuroendocrine regulation of long-term pair maintenance in the monogamous zebra finch.

This article is part of a Special Issue "SBN 2014". Understanding affiliative behavior is critical to understanding social organisms. While affiliative behaviors are present across a wide range of taxa and contexts, much of what is known about the neuroendocrine regulation of affiliation comes from studies of pair-bond formation in prairie voles. This leaves at least three gaps in our current knowledge. First, little is known about long-term pair-bond maintenance. Second, few studies have examined non-mammalian systems, even though monogamy is much more common in birds than in mammals. Third, the influence of breeding condition on affiliation is largely unknown. The zebra finch (Taeniopygia guttata) is an excellent model system for examining the neuroendocrine regulation of affiliative behaviors, including the formation and maintenance of a long-term pair bond. Zebra finches form genetically monogamous pair bonds, which they actively maintain throughout the year. The genomic and neuroanatomical resources, combined with the wealth of knowledge on the ecology and ethology of wild zebra finches, give this model system unique advantages to study the neuroendocrine regulation of pair bonding. Here, we review the endocrinology of opportunistic breeding in zebra finches, the sex steroid profiles of breeding and non-breeding zebra finches (domesticated and wild), and the roles of sex steroids and other signaling molecules in pair-maintenance behaviors in the zebra finch and other monogamous species. Studies of zebra finches and other songbirds will be useful for broadly understanding the neuroendocrine regulation of affiliative behaviors, including pair bonding and monogamy.

Acute and chronic effects of an aromatase inhibitor on pair-maintenance behavior of water-restricted zebra finch pairs.

Zebra finches are highly social songbirds that maintain life-long monogamous pair-bonds. They rely heavily upon these pair-bonds to survive their ever-changing and unpredictable habitat in the Australian desert. These pair-bonds are maintained via a large repertoire of affiliative behaviors that for most of an individual's life are predominately associated with pair maintenance. Water restriction reduces circulating testosterone levels in male zebra finches and the size of the ovary and oviduct in female zebra finches, but water restriction has little or no effects on pair-maintenance behaviors and local levels of testosterone and estradiol in behaviorally-relevant brain regions. These data suggest that in water-restricted zebra finches, local synthesis of testosterone and estradiol in the brain may support the expression of pair-maintenance behaviors. Here, we directly test whether pair-maintenance behaviors are regulated by estradiol, acting via non-genomic or genomic mechanisms, in water-restricted (i.e., non-breeding) zebra finches. In two experiments, subjects were treated with an aromatase inhibitor (fadrozole) either acutely or chronically, and a variety of pair-maintenance behaviors were quantified. Additionally, we quantified the effect of acute fadrozole treatment on brain and circulating estradiol and testosterone levels. Acute fadrozole administration rapidly decreased estradiol levels in the circulation and brain of males and also rapidly increased testosterone levels in the circulation and brain of both males and females. However, neither the acute nor chronic fadrozole treatment decreased pair-maintenance behaviors. In one case, acute fadrozole treatment promoted affiliation. These data suggest that pair-maintenance behavior in non-breeding zebra finches is not promoted by estradiol acting via either non-genomic or genomic mechanisms.

Fasting increases aggression and differentially modulates local and systemic steroid levels in male zebra finches.

Aggression enables individuals to obtain and retain limited resources. Studies of the neuroendocrine regulation of aggression have focused on territorial and reproductive contexts. By contrast, little is understood concerning the neuroendocrine regulation of aggression over other resources, such as food. Here, we developed a paradigm to examine the role of steroids in food-related aggression. In groups of male zebra finches, a 6-hour fast decreased body mass and increased aggressive interactions among subjects that competed for a point source feeder. Fasting also dramatically altered circulating steroid levels by decreasing plasma testosterone but not estradiol (E2). By contrast, both plasma corticosterone and dehydroepiandrosterone (DHEA) concentrations were elevated with fasting. Interestingly, short-term access to food (15 minutes) after fasting normalized circulating steroid levels. Fasting increased corticosterone levels in a wide range of peripheral tissues but increased DHEA levels specifically in adrenal glands and liver; these effects were quickly normalized with refeeding. DHEA can be metabolized within specific brain regions to testosterone and E2, which promote the expression of aggression. We measured E2 in microdissected brain regions and found that fasting specifically increased local E2 levels in 3 regions: the periaqueductal gray, ventral tegmental area, and ventromedial nucleus of the hypothalamus. These regions are part of the vertebrate social behavior network and regulate the expression of aggression. Together, these data suggest that fasting stimulates secretion of DHEA from the adrenals and liver and subsequent conversion of DHEA to E2 within specific brain regions, to enable individuals to compete for limited food resources.

Effects of water restriction on reproductive physiology and affiliative behavior in an opportunistically-breeding and monogamous songbird, the zebra finch.

Wild zebra finches form long-term monogamous pair-bonds that are actively maintained year-round, even when not in breeding condition. These desert finches are opportunistic breeders, and breeding is highly influenced by unpredictable rainfall. Their high levels of affiliation and complex breeding patterns make zebra finches an excellent model in which to study the endocrine regulation of affiliation. Here, we compared zebra finch pairs that were provided with water ad libitum (control) or water restricted. We examined (1) reproductive physiology, (2) pair-maintenance behaviors in several contexts, and (3) circulating and brain steroid levels. In females, water restriction profoundly reduced largest ovarian follicle size, ovary size, oviduct size, and egg laying. In males, water restriction had no effect on testes size but decreased systemic testosterone levels. However, in the hypothalamus, local testosterone and estradiol levels were unaffected by water restriction in both sexes. Systemic and local levels of the androgen precursor dehydroepiandrosterone (DHEA) were also unaffected by water restriction. Lastly, in three different behavioral paradigms, we examined a variety of pair-maintenance behaviors, and none were reduced by water restriction. Taken together, these correlational data are consistent with the hypothesis that local production of sex steroids in the brain promotes the expression of pair-maintenance behaviors in non-breeding zebra finches.