Behavioral and physiological responses to experimentally elevated testosterone in female dark-eyed juncos (Junco hyemalis carolinensis)

Testosterone mediates the expression of many fitness-related traits in male vertebrates and is thought to account for numerous sex differences in trait expression. Testosterone is also secreted by females; however, far less is known regarding its effects on female physiology and behavior. Using a bird species in which the effects of testosterone on males are well characterized, the dark-eyed junco (Junco hyemalis), we tested whether an increase in exogenous testosterone in females would alter the phenotypic expression of a suite of behavioral and physiological traits. We found that increased testosterone levels in female dark-eyed juncos led to decreased cell-mediated immune function and increased intrasexual aggression, hypothalamo-pituitary-adrenal (HPA) axis responsiveness, baseline corticosterone and corticosterone-binding globulin (CBG) levels. Furthermore, immunosuppression following testosterone implantation was negatively correlated with total and free testosterone but did not appear to be related to either total or free corticosterone. These results demonstrate that the phenotypic impact of elevated testosterone is not confined to males in dark-eyed juncos, and that the impact in adults can be similar in males and females. We discuss these results in the context of potential endocrine-immune interactions and the evolution of sexual dimorphism.     

Recent postglacial range expansion drives the rapid diversification of a songbird lineage in the genus Junco

Pleistocene glacial cycles are thought to have played a major role in the diversification of temperate and boreal species of North American birds. Given that coalescence times between sister taxa typically range from 0.1 to 2.0 Myr, it has been assumed that diversification occurred as populations were isolated in refugia over long periods of time, probably spanning one to several full glacial cycles. In contrast, the rapid postglacial range expansions and recolonization of northern latitudes following glacial maxima have received less attention as potential promoters of speciation. Here we report a case of extremely rapid diversification in the songbird genus Junco as a result of a single continent-wide range expansion within the last 10 000 years. Molecular data from 264 juncos sampled throughout their range reveal that as the yellow-eyed junco (Junco phaeonotus) of Mesoamerica expanded northward following the last glacial maximum, it speciated into the dark-eyed junco (Junco hyemalis), which subsequently diversified itself into at least five markedly distinct and geographically structured morphotypes in the USA and Canada. Patterns of low genetic structure and diversity in mitochondrial DNA and amplified fragment length polymorphism loci found in dark-eyed juncos relative to Mesoamerican yellow-eyed juncos provide support for the hypothesis of an expansion from the south, followed by rapid diversification in the north. These results underscore the role of postglacial expansions in promoting diversification and speciation through a mechanism that represents an alternative to traditional modes of Pleistocene speciation.     

Winter social behaviour of dark-eyed juncos: Communication,social organization,and ecological implications

This paper reports on winter social behaviour in free-living and captive flocks of dark-eyed juncos (Junco hyemalis). Visual and vocal signals used by flocked juncos are analysed structurally and functionally. An examination is made of the frequency and form of agonistic behaviour in several social and environmental contexts in an attempt to provide insight into the relationship between junco communication and social organization. A system of classification of signals, based upon the meaning of cues to the signaller, is proposed. Evidence is given to suggest that the behaviour of subordinate, rather than of dominant, individuals controls the operation of the social system in junco flocks. Some ecological implications of junco social behaviour are discussed.     

Winter fattening in the dark-eyed junco: plasticity and possible interaction with migration trade-offs

Although fat often supplies the major source of metabolic fuel during winter fasts of birds, this critical life-history trait is little studied by ecologists. In the dark-eyed junco Junco hyemalis, we have in a series of studies investigated the extent of plasticity in the winter fat reserve. Earlier (Rogers et al. 1993), we reported (1) a highly variable pattern of geographic variation in the winter fat reserve of junco populations in eastern North America, (2) disappearance of statistically significant interpopulation variation after experimental displacement to a common latitude, and (3) post-displacement temporal variation in the fat reserve. In analyses reported here, recent temperature, recent snowfall (a measure of short-term predictability of resources), season (perhaps reflecting continued exposure to unpredictable resources) and daylength explained spatial variation in the fat store. Recent temperature explained temporal variation in the fat reserves of groups of displaced juncos. These results suggest that platticity in a life-history trait has evolved in an uncertain winter environment. Through environment-dependent fattening, the costs of fat can be avoided during warm periods and at locations where fat confers little benefit, whereas benefits of fat can be quickly gained if weather conditions become harsh and snowfall might restrict food. Three types of winter fatteners probably exist among birds: responders (fatten in response to the proximate environment), predictors (fatten in anticipation of long-term environmental conditions), and responder-predictors (combination of both types of regulation). Because dark-eyed juncos select different winter latitudes as they age, we hypothesize that the nonbreeding component of the life-history of juncos includes the co-adapted plastic traits of winter fattening and post-breeding migration. Life-history theory can apparently explain important traits related to fitness in the nonbreeding period.     

Migratory dark-eyed juncos, Junco hyemalis, have better spatial memory and denser hippocampal neurons than nonmigratory conspecifics

The evolution of migration in an animal population produces a suite of physiological, behavioural and cognitive adaptations. Migratory birds, in particular, require the ability to return annually to breeding and wintering sites after long journeys, and thus might be predicted to have evolved enhanced spatial memory. In a comparison of two sparrow subspecies that co-occur in winter, the migratory subspecies (dark-eyed junco, Junco hyemalis hyemalis) performed better than the nonmigratory subspecies (J. h. carolinensis) on a room-scale spatial memory test. The migratory juncos also had more densely packed hippocampal neurons than did nonmigrants. Among nonmigrants, we looked for hippocampal differences between birds that occupied two home ranges annually and those that remained on their breeding territory year-round, to determine whether migration, per se, is related to neuroanatomical differences. However, we were unable to reach any conclusions because of low statistical power. A denser hippocampus could be the basis for better spatial memory in migrant juncos. Further testing of spatial memory on a landscape scale is needed to strengthen this argument and to understand cognitive differences between migrants and nonmigrants. Copyright 2003 Published by Elsevier Science Ltd on behalf of The Association for the Study of Animal Behaviour.      

Boldness behavior and stress physiology in a novel urban environment suggest rapid correlated evolutionary adaptation

Novel or changing environments expose animals to diverse stressors that likely require coordinated hormonal and behavioral adaptations. Predicted adaptations to urban environments include attenuated physiological responses to stressors and bolder exploratory behaviors, but few studies to date have evaluated the impact of urban life on codivergence of these hormonal and behavioral traits in natural systems. Here, we demonstrate rapid adaptive shifts in both stress physiology and correlated boldness behaviors in a songbird, the dark-eyed junco, following its colonization of a novel urban environment. We compared elevation in corticosterone (CORT) in response to handling and flight initiation distances in birds from a recently established urban population in San Diego, California to birds from a nearby wildland population in the species' ancestral montane breeding range. We also measured CORT and exploratory behavior in birds raised from early life in a captive common garden study. We found persistent population differences for both reduced CORT responses and bolder exploratory behavior in birds from the colonist population, as well as significant negative covariation between maximum CORT and exploratory behavior. Although early developmental effects cannot be ruled out, these results suggest contemporary adaptive evolution of correlated hormonal and behavioral traits associated with colonization of an urban habitat.     

Social status does not affect resting metabolic rate in wintering dark-eyed junco (Junco hyemalis)

Studies of wintering birds have demonstrated a correlation between social rank and energy expenditures. It is assumed that dominance is energetically costly because of increased activity, possibly caused by elevated androgen levels. As winter acclimatization leads to an increase in metabolic rate, maintaining dominance status in a cold climate can be a substantial challenge. We measured resting metabolic rates in dominant and subordinate dark-eyed juncos (Junco hyemalis) living in small groups in a controlled winter environment. We found no significant effect of social rank when controlling for body size. It has been shown previously that high testosterone levels during the nonbreeding season can lead to higher body conductance, fat loss, and higher nocturnal body temperature. A hypothesis explaining our result is that for juncos it is preferable to maintain low androgen levels during winter and to maintain social rank using a mechanism other than higher agonistic activity.     

Testosterone and opioids interact to regulate feeding in a male migratory songbird.

A male migratory songbird (dark-eyed junco, Junco hyemalis) was used as a model for studies on the influence of testosterone (T) on feeding, and on interactive effects on this behavior between T and the opioid antagonist naloxone hydrochloride (Nal). Administered chronically to birds exposed to nonstimulating photoperiods, T increased food intake by 30-58% without altering the body mass, the fat index, or the standard metabolic rate. An intramuscular injection of Nal decreased feeding temporarily in a dose-related manner. T-treated juncos exhibited a decreased sensitivity to the anorexic influence of Nal administration, demonstrating that T interacts with opioids to control food consumption. Neuroendocrine mechanisms that potentially account for this interaction are discussed.     

Temporal genetic structure and relatedness in the Tufted Duck Aythya fuligula suggests limited kin association in winter

Conspecific aggregation of waterfowl in winter is a common example of animal flocking behaviour, yet patterns of relatedness and temporal substructure in such social groups remain poorly understood even in common species. A previous study based on mark‐recapture data showed that Tufted Ducks Aythya fuligula caught on the same day were re‐caught together in subsequent winters more often than expected by chance, suggesting stable assortments of ‘socially familiar’ individuals between wintering periods. The genetic relationships within these social groups were not clear. Based on 191 individuals genotyped at 10 microsatellite markers, we investigated the temporal genetic structure and patterns of relatedness among wintering Tufted Ducks at Lake Sempach, Switzerland, in two consecutive winters. We found no evidence of genetic differentiation between temporal groups within or between winters. The average levels of relatedness in temporal groups were low and not higher than expected in random assortments of individuals. However, Mantel tests performed for each sex separately revealed significant negative correlations between the pairwise relatedness coefficients and the number of days between the capture dates of pairs of wintering Tufted Duck in males and females. This pattern suggests the presence of a small number of co‐migrating same‐sex sibling pairs in wintering flocks of Tufted Ducks. Our findings provide one of the first genetic analyses of a common duck species outside the breeding season and contribute to the understanding of social interactions in long‐distance migratory birds.     

Corticosterone secretion patterns prior to spring and autumn migration differ in free-living barn swallows (Hirundo rustica L.)

Recent studies of long-distance migratory birds show that behavioural and physiological changes associated with predictable or unpredictable challenges during the annual cycle are distinctively regulated by hormones. Corticosterone is the primary energy regulating hormone in birds. Corticosterone levels are elevated during stresses but they are also modulated seasonally according to environmental conditions and life-history demands. We measured the baseline and stress-induced levels of corticosterone in the barn swallow (Hirundo rustica L.) just before spring and autumn migrations in South Africa and Finland, respectively. Barn swallows completing their pre-breeding moult had low body condition (residual body mass) and high baseline corticosterone levels in the wintering grounds. In contrast, baseline corticosterone levels in Finland were low and not related to residual mass. These data contradict the first prediction of the migration modulation hypothesis (MMH) by showing no association with baseline corticosterone levels and pre-migratory fuelling. Yet, the adrenocortical response to the capture and handling stress was notably blunted in South Africa compared to a strong response in Finland. Further, individuals that had started fuelling in Finland showed a reduced response to the handling stress. Taken together, elevated baseline corticosterone levels and high residual mass may blunt the adrenocortical response in long-distance migrants and aerial feeders such as the barn swallow. This observation lends support to the second prediction of the MMH.     

Consequences of elevating plasma testosterone in females of a socially monogamous songbird: evidence of constraints on male evolution?

To explore whether selection for testosterone-mediated traits in males might be constrained by costs of higher testosterone to females, we examined the effects of experimental elevation of plasma testosterone on physiological, reproductive, and behavioral parameters in a female songbird, the dark-eyed junco (Junco hyemalis). We used subcutaneous implants to elevate testosterone (T) in captive and free-living female juncos. In captive birds, we measured the effects of high T on body mass, feather molt, and brood patch formation. In the field, we monitored its effects on the timing of egg laying, clutch size, egg size, egg steroid levels, incubation, and nest-defense behavior. Females implanted with testosterone (T-females) had significantly higher circulating levels of testosterone than did control females (C-females). Captive T-females had lower body mass, were less likely to develop brood patches, and delayed feather molt relative to C-females. Among free-living females, the interval between nest completion and appearance of the first egg was longer for T-females than for C-females and egg yolk concentrations of testosterone were higher, but there were no significant differences in estradiol levels, clutch size, or egg size. Incubation and nest defense behavior were also similar between T- and C-females. Our results suggest that selection on males for higher testosterone might initially lead to a correlated response in females producing changes in body mass and feather molt, both of which could be detrimental. Other possible female responses would be delayed onset of reproduction, which might reduce reproductive success, and higher yolk testosterone, which might have either positive or negative effects on offspring development. We found no reason to expect reduced parental behavior by females as a negative fitness consequence of selection for higher testosterone in males.     

Effects of captivity and testosterone on the volumes of four brain regions in the dark-eyed junco (Junco hyemalis)

This study investigates the effects of captivity and testosterone treatment on the volumes of brain regions involved in processing visual and spatial information in adult dark-eyed juncos (Junco hyemalis). We treated captive and free-living male juncos with either testosterone-filled or empty implants. Captive juncos had a smaller hippocampal formation (HF) (both in absolute volume and relative to telencephalon) than free-living birds, regardless of hormone treatment. Testosterone-treated males (both captive and free-living) had a smaller telencephalon and nucleus rotundus, but not a smaller HF or ectostriatum, than controls. We found that free-living testosterone-treated males had larger home ranges than free-living controls in agreement with earlier experiments, but we found no corresponding difference in HF volume. We discuss the implications of the effect of captivity on HF volume for past and future laboratory experiments.     

Interspecific dominance and the foraging behaviour of juncos

We observed mixed groups of dark-eyed juncos (Junco hyemalis) and grey-headed juncos (Junco caniceps) at baited sites in northern Arizona during the non-breeding season. In interspecific and inter-racial conflicts, J. caniceps dorsalis was dominant to J. caniceps caniceps and to two races of dark-eyed juncos. Junco caniceps dorsalis also fed significantly faster than any of the other juncos. For both species, feeding rates were approximately the same in large and small mixed-species groups, though in larger groups, individual grey-headed juncos won conflicts at a higher rate and individual dark-eyed juncos lost conflicts at a higher rate. Also, dark-eyed juncos fed at a significantly lower rate in groups comprised mostly of grey-headed juncos than in groups of similar size but composed mostly of conspecifics. Residency times and recapture probabilities were similar for the two species, suggesting little difference in over-winter survival.     

Carry‐over effects provide linkages across the annual cycle of a Neotropical migratory bird,the Louisiana Waterthrush Parkesia motacilla

Population limitation models of migratory birds have sought to include impacts from events across the full annual cycle. Previous work has shown that events occurring in winter result in some individuals transitioning to the breeding grounds earlier or in better physical condition than others, thereby affecting reproductive success (carry‐over effects). However, evidence for carry‐over effects from breeding to wintering grounds has been shown less often. We used feather corticosterone (CORT_f) levels of the migratory Louisiana Waterthrush Parkesia motacilla as a measure of the physiological state of birds at the time of moult on the breeding territory to investigate whether carry‐over effects provide linkages across the annual cycle of this stream‐obligate bird. We show that birds arriving on wintering grounds with lower CORT_f scores, indicating reduced energetic challenges or stressors at the time of moult, occupied higher quality territories, and that these birds then achieved a better body condition during the overwinter period. Body condition, in turn, was important in determining whether adult birds returned the following winter, with birds in better condition returning at higher rates. Together these data suggest a carry‐over effect from the breeding grounds to the wintering grounds that is further extended with respect to annual return rates. Very few other studies have linked conditions during the previous breeding season with latent effects during the subsequent overwintering period or with annual survival. This study shows that the effects of variation in energetic challenges or stressors can potentially carry over from the natal stream and accumulate over more than one life‐history period before being manifested in reduced survival. This is of particular relevance to models of population limitation in migratory birds.     

Female ornamentation and male mate choice in dark-eyed juncos

Traits that enhance attractiveness in one sex may or may not influence attractiveness in the other. In the dark-eyed junco, Junco hyemalis, outer tail feathers of males and females are all or partly white and form a sharp contrast with the bird's mostly grey plumage. The amount of white in these feathers (‘tail white’) is greater in males than in females and, as we report here, is greater in birds that have completed a second prebasic moult than in yearlings. During courtship, male juncos spread their tails, revealing their tail white, and a previous experiment has shown that males with experimentally enhanced tail white are more attractive to females. To determine whether females with experimentally enhanced tail white would be preferred by males, we clipped and replaced tail feathers of females, creating a control group with low to natural levels of tail white and an enhanced group with high levels. We tested preference in a mate choice apparatus like that used previously and found that males courted both control and enhanced females and displayed individual preferences but showed no collective preference for members of either category of females. Because we found neither a preference for trait values that indicate greater age/experience (experimentally enhanced females) nor a preference for less male-like appearance (control females), our results are inconsistent with a role for male mate choice in the maintenance of tail white in females. Female tail white may be subject to selection in another context or persist owing to a genetic correlation between the sexes. Regardless, the sexes apparently prefer different trait values, which suggests that preferences are expressed independently in males and females.     

Sex and migratory strategy influence corticosterone levels in winter-grown feathers,with positive breeding effects in a migratory pelagic seabird

To overcome unpredictable stressful transitory events, animals trigger an allostatic response involving the hypothalamic–pituitary–adrenal cortex. This hormonal response, which involves the release of glucocorticoids which in turn mediate between the main physiological mechanisms that regulate the energetic demands and resource allocation trade-off with behavioural responses to environmental perturbations and may ultimately lead to variation in fitness. We have used the Cory’s shearwater Calonectris borealis, a sexually dimorphic pelagic seabird with a partial migratory strategy, as a model bird species to analyse a number of traits related to the stress response. We investigated whether the activation of a stressful response, mediated by corticosterone, during the wintering period (1) correlated with the previous breeding success, (2) was affected by the migratory behaviour of male birds and (3) had consequences in the fitness of the birds. Corticosterone levels in feathers grown overwinter were analysed in 61 adult birds during three consecutive migratory periods (2009–2012) and in 14 immature birds in the wintering period 2010–2011. Moreover, the levels of corticosterone were analysed in experimental birds which were freed from their reproductive duties and compared with control birds which raised fledglings to the end of the breeding period. The results show that the levels of corticosterone were sex dependent, differed between years and were affected by the migratory strategy performed by the birds. The activation of the stressful response over the wintering period generated residual carry-over effects that positively affected the reproductive output in the subsequent breeding stage, a phenomenon previously undescribed in a long-lived pelagic seabird. Our study provides evidence that the analysis of corticosterone from feathers is a useful tool to evaluate carry-over effects in birds far away from breeding sites, opening new possibilities for future studies in this field.     

Testosterone in females: mediator of adaptive traits, constraint on sexual dimorphism, or both?

When selection on males and females differs, the sexes may diverge in phenotype. Hormones serve as a proximate regulator of sex differences by mediating sex-biased trait expression. To integrate these perspectives, we consider how suites of traits mediated by the same hormone in both sexes might respond to selection. In male birds, plasma testosterone (T) varies seasonally and among species according to mating system. When elevated experimentally, it is known to enhance some components of fitness and to decrease others. We report that female T also varies seasonally and co-varies with male T. Female T is higher in relation to male T in sexually monomorphic species and is higher absolutely in females of species with socially monogamous mating systems, which suggests adaptation. We also consider the effect of experimentally elevated T on females and whether traits are sensitive to altered T. We hypothesize that sensitive traits could become subject to selection after a natural change in T and that traits with opposing fitness consequences in males and females could constrain dimorphism. Results from birds, including the dark-eyed junco (Junco hyemalis), reveal many sensitive traits, some of which appear costly and may help to account for observed levels of sexual dimorphism.     

Annual Winter Site Fidelity of Barrow's Goldeneyes in the Pacific

Coastal regions on the Pacific north coast of North America provide important wintering habitat for many species of sea ducks. Although winter range and habitat preferences are well described for most species, fidelity to coastal wintering sites is generally undocumented. Fidelity is an important factor necessary for understanding interactions with coastal developments and activities and corresponding management strategies. We used data from Barrow's goldeneyes (Bucephala islandica), a sea duck that winters predominantly in nearshore habitats along the Pacific north coast, to investigate inter-annual fidelity to, and intra-annual fidelity within, coastal wintering sites. Between 2006 and 2015, we marked goldeneyes on breeding, molting, and wintering sites with satellite transmitters. We retained 4,931 locations in coastal habitats from 221 goldeneyes across 4 coastal regions for our analyses. These birds demonstrated high inter-annual fidelity to coastal wintering sites; 75% of selected wintering sites were within 29 km of sites used the previous winter. Inter-annual fidelity to wintering sites was similar between sex and age classes but differed by coastal region. Goldeneyes from southcentral Alaska, USA, expressed greater inter-annual fidelity relative to birds from northern or southern British Columbia, Canada, and southeast Alaska. Goldeneyes also expressed high intra-annual fidelity within wintering sites, with 75% of individuals averaging within-season movements of ≤9 km. Intra-annual fidelity was lesser for female than male goldeneyes but did not differ between hatch-year and after-hatch-year birds. We found regional variation in intra-annual fidelity, with goldeneyes from southcentral Alaska expressing greater intra-annual fidelity compared to birds from other regions. High inter- and intra-annual winter site fidelity by Barrow's goldeneyes suggests that, at a population level, habitat use is predictable and can be used to inform risk assessment or to evaluate factors affecting habitat choice. Also, low dispersal among wintering sites suggests that recovery from population perturbations, whether caused by natural or anthropogenic events, will be protracted. © 2019 The Wildlife Society.     

Photoperiod and testosterone independently affect vocal control region volumes in adolescent male songbirds

Previously, we found that, unlike adults, adolescent male dark-eyed juncos (Junco hyemalis) maintained large Area X volumes despite having low plasma testosterone concentrations. Other studies indicate that photoperiod may act independently of testosterone to modulate vocal control region (VCR) volumes in adult songbirds. In the present study, we investigated the effects of testosterone and photoperiod on the volumes of four VCRs in adolescent male juncos. To test the hypothesis that VCR volumes in these males are testosterone independent, we treated birds exposed to short days with testosterone and later compared their VCR volumes with those of birds exposed to short days without testosterone. To examine whether photoperiod alone could affect VCR volumes independent of testosterone, we measured these volumes in photorefractory birds exposed to long photoperiod without testosterone. Administering testosterone induced singing, yet increased the volume of only one VCR, the robust nucleus of the anterior neostriatum (RA). In contrast, long photoperiod increased several VCR volumes (Area X, higher vocal center, and RA) despite low testosterone levels, but did not induce singing. Our results suggest a limited role for testosterone, but an important role for photoperiod, in controlling VCR volumes in adolescent male juncos. In addition, the results demonstrate that singing behavior can be induced in adolescent males without a concomitant increase in most VCR volumes. © 1998 John Wiley & Sons, Inc. J Neurobiol 36: 550–558, 1998     

Seasonal and diel transitions in physiology and behavior in the migratory dark-eyed junco

Body mass, fat stores, activities of lipogenic and lipolytic enzymes, and plasma corticosterone were measured throughout seasonal and diel transitions from fall through spring encompassing the non-migratory stages of early and mid winter, the prealternate molt, and the spring migratory stage in captive dark-eyed juncos to determine the physiological mechanisms underlying adaptations for migration. On a seasonal basis, lipid enzymes and corticosterone varied little throughout the stages even though the birds underwent dramatic alterations in mass, fat deposition, behavior, and activation of the reproductive axis. By contrast, diel changes were found in lipogenesis, lipolysis, muscle lipoprotein lipase, and plasma corticosterone when comparing birds in the two phases of spring migration--active flight and resting, as during times of stopover. In these two phases of migration, coordination of the lipogenic and lipolytic systems appear to maximize storage of fatty acids during rest and delivery/utilization during flight. Diel patterns of corticosterone revealed fairly consistent peaks during the night time (23:00) throughout the nonmigratory period. The profile of this pattern altered during the migratory period with variation between the flight and resting phases. In sum, the results from these captive studies offer a new approach for studying the regulation of migratory physiology in free-living birds.