An Aggressive Interaction Rapidly Increases Brain Androgens in a Male Songbird during the Non-breeding Season.

Aggression is a crucial behavior that impacts access to limited resources in different environmental contexts. Androgens synthesized by the gonads promote aggression during the breeding season. However, aggression can be expressed during the non-breeding season, despite low androgen synthesis by the gonads. The brain can also synthesize steroids ("neurosteroids"), including androgens, which might promote aggression during the non-breeding season. Male song sparrows, Melospiza melodia, are territorial year-round and allow the study of seasonal changes in the steroid modulation of aggression. Here, we quantified steroids following a simulated territorial intrusion (STI) for 10 min in wild adult male song sparrows during the breeding and non-breeding seasons. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we examined 11 steroids: pregnenolone, progesterone, corticosterone, dehydroepiandrosterone, androstenedione, testosterone, 5α-dihydrotestosterone, 17ß-estradiol, 17α-estradiol, estriol, and estrone. Steroids were measured in blood and 10 microdissected brain regions that regulate social behavior. In both seasons, STI increased corticosterone in the blood and brain. In the breeding season, STI had no rapid effects on androgens or estrogens. Intriguingly, in the non-breeding season, STI increased testosterone and androstenedione in several behaviorally relevant regions, but not in the blood, where androgens remained non-detectable. Also in the non-breeding season, STI increased progesterone in the blood and specific brain regions. Overall, rapid socially modulated changes in brain steroid levels are more prominent during the non-breeding season. Brain steroid levels vary with season and social context in a region-specific manner and suggest a role for neuroandrogens in aggression during the non-breeding season.

White-throated sparrow (Zonotrichia albicollis) liver and pectoralis flight muscle transcriptomic changes in preparation for migration.

Migratory songbirds undertake challenging journeys to reach their breeding grounds each spring. They accomplish these nonstop flapping feats of endurance through a suite of physiological changes, including the development of substantial fat stores and flight muscle hypertrophy and an increased capacity for fat catabolism. In addition, migratory birds may show large reductions in organ masses during flight, including the flight muscle and liver, which they must rapidly rebuild during their migratory stopover before replenishing their fat stores. However, the molecular basis of this capacity for rapid tissue remodeling and energetic output has not been thoroughly investigated. We performed RNA-sequencing analysis of the liver and pectoralis flight muscle of captive white-throated sparrows in experimentally photostimulated migratory and nonmigratory condition to explore the mechanisms of seasonal change to metabolism and tissue mass regulation that may facilitate these migratory journeys. Based on transcriptional changes, we propose that tissue-specific adjustments in preparation for migration may alleviate the damaging effects of long-duration activity, including a potential increase to the inflammatory response in the muscle. Furthermore, we hypothesize that seasonal hypertrophy balances satellite cell recruitment and apoptosis, while little evidence appeared in the transcriptome to support myostatin-, insulin-like growth factor 1-, and mammalian target of rapamycin-mediated pathways for muscle growth. These findings can encourage more targeted molecular studies on the unique integration of pathways that we find in the development of the migratory endurance phenotype in songbirds.NEW & NOTEWORTHY Migratory songbirds undergo significant physiological changes to accomplish their impressive migratory journeys. However, we have a limited understanding of the regulatory mechanisms underlying these changes. Here, we explore the transcriptomic changes to the flight muscle and liver of white-throated sparrows as they develop the migratory condition. We use these patterns to develop hypotheses about metabolic flexibility and tissue restructuring in preparation for migration.

Physiological status of House Sparrows (Passer domesticus) along an ozone pollution gradient.

Mexico City is one of the most polluted cities in the world, and one in which air contamination is considered a public health threat. Numerous studies have related high concentrations of particulate matter and ozone to several respiratory and cardiovascular diseases and a higher human mortality risk. However, almost all of those studies have focused on human health outcomes, and the effects of anthropogenic air pollution on wildlife species is still poorly understood. In this study, we investigated the impacts of air pollution in the Mexico City Metropolitan Area (MCMA) on house sparrows (Passer domesticus). We assessed two physiological responses commonly used as biomarkers: stress response (the corticosterone concentration in feathers), and constitutive innate immune response (the concentration of both natural antibodies and lytic complement proteins), which are non-invasive techniques. We found a negative relationship between the ozone concentration and the natural antibodies response (p = 0.003). However, no relationship was found between the ozone concentration and the stress response or the complement system activity (p > 0.05). These results suggest that ozone concentrations in air pollution within MCMA may constrain the natural antibody response in the immune system of house sparrows. Our study shows, for the first time, the potential impact of ozone pollution on a wild species in the MCMA presenting the Nabs activity and the house sparrow as suitable indicators to assess the effect of air contamination on the songbirds.

Canalised and plastic components of melanin-based colouration: a diet-manipulation experiment in house sparrows.

Whether melanin-based plumage colouration accurately reflects a bird's quality is still controversial. To better understand potential mechanisms behind the observed variation in plumage colouration, we shifted our attention from a high-level expression of colour to low-level physiological phenomena by targeting the microstructure and pigment content of the feather. In a well-studied model system, the house sparrow (Passer domesticus), we combined an experimental manipulation of birds' physiological condition and availability of resources that are key to the production of the studied colouration (phenylalanine and tyrosine (PT). We found that feathers from sparrows fed with the control diet had noticeably lower values of brightness, suggesting a higher quality of the ornamental "blackness" in comparison to those sampled from birds fed with a PT-reduced diet. Electron paramagnetic resonance (EPR) spectroscopy detected higher melanin concentrations in samples from the control than the PT-reduced group. Our multi-level analysis excluded mechanisms such as barbule density and melanosomes' distribution, clearly pointing to the finest-level proxy of colour: the concentration of melanin in melanosomes themselves. Despite melanins being manufactured by birds endogenously, the efficiency of melanogenesis can be noticeably limited by diet. As a result, the birds' plumage colouration is affected, which may entail consequences in social signalling.

Experimental Exposure to Tebuconazole Affects Metabolism and Body Condition in a Passerine Bird, the House Sparrow (Passer domesticus).

Triazole compounds are among the most widely used fungicides in agroecosystems to protect crops from potential fungal diseases. Triazoles are suspected to have an impact on nontarget species due to their interactions with nonfungal sterol synthesis, and wild birds are likely to be contaminated by triazole fungicides because many of them live in agroecosystems. We experimentally tested whether exposure to environmental concentrations of a triazole could alter key integrative traits (metabolic rates and body condition) of an agroecosystem sentinel species, the house sparrow (Passer domesticus). Wild-caught adult sparrows were maintained in captivity and exposed (exposed group) or not (control group) for 7 continuous months to tebuconazole through drinking water. The metabolic rates of exposed and control sparrows were then measured at two different temperatures (12 °C and 25 °C), which correspond, respectively, to the thermoregulation and thermoneutrality temperatures of this species. We found that exposed sparrows had lower resting metabolic rates (i.e., measured at thermoneutrality, 25 °C) than controls. However, the thermoregulatory metabolic rates (i.e., measured at 12 °C) did not differ between exposed and control sparrows. Although the body mass and condition were not measured at the beginning of the exposure, sparrows at the time of the metabolic measurements 7 months after the onset of such exposure had a higher body condition than controls, supporting further the idea that tebuconazole affects metabolic functions. Our study demonstrates for the first time that the use of tebuconazole can alter metabolism and could potentially lead to adverse effects in birds. Environ Toxicol Chem 2022;41:2500-2511. © 2022 SETAC.

The Effect of a Combined Fast and Chronic Stress on Body Mass, Blood Metabolites, Corticosterone, and Behavior in House Sparrows (Passer domesticus).

One aspect of the Reactive Scope Model is wear-and-tear, which describes a decrease in an animal's ability to cope with a stressor, typically because of a period of chronic or repeated stressors. We investigated whether wear-and-tear due to chronic stress would accelerate a transition from phase II to phase III of fasting. We exposed house sparrows (Passer domesticus) to three weeks of daily fasts combined with daily intermittent repeated acute stressors to create chronic stress, followed by two weeks of daily fasts without stressors. We measured circulating glucose, ß-hydroxybutyrate (a ketone), and uric acid in both fasted and fed states. We expected birds to be in phase II (high fat breakdown) in a fasted state, but if wear-and-tear accumulated sufficiently, we hypothesized a shift to phase III (high protein breakdown). Throughout the experiment, the birds exhibited elevated ß-hydroxybutyrate when fasting but no changes in circulating uric acid, indicating that a transition to phase III did not occur. In both a fasted and fed state, the birds increased glucose mobilization throughout the experiment, suggesting wear-and-tear occurred, but was not sufficient to induce a shift to phase III. Additionally, the birds exhibited a significant decrease in weight, no change in corticosterone, and a transient decrease in neophobia with chronic stress. In conclusion, the birds appear to have experienced wear-and-tear, but our protocol did not accelerate the transition from phase II to phase III of fasting.

Novel objects alter immediate early gene expression globally for ZENK and regionally for c-Fos in neophobic and non-neophobic house sparrows.

Neophobia - an animal's reluctance to approach novel objects, try new foods, or explore unfamiliar environments - affects whether animals can adapt to new environments and exploit novel resources. However, despite its importance, the neurobiological mechanisms underlying this personality trait are poorly understood. In this study, we examined regional brain activity using the expression of two immediate early genes (IEGs), ZENK and c-Fos, in response to novel objects or control conditions in captive house sparrows (Passer domesticus, n = 22). When exposed to novel objects, we predicted that we would see differential IEG activity in brain regions involved in regulating stress and emotion (hippocampus, medial ventral arcopallium, lateral septum), reward and learning (striatum), and executive function (NCL) between neophobic and non-neophobic individuals. To classify birds by phenotype, we used behavior trials that tested willingness to approach a food dish in the presence of several different novel objects, habituation to one novel object, and willingness to try several different novel foods. We then exposed birds to a new novel object or a control condition and assessed protein expression of two IEGs in neophobic vs non-neophobic individuals after this final exposure. An analysis of average sparrow feeding times in the presence of novel objects showed a bimodal distribution of neophobia behavior. There was also high repeatability of individual novel object responses, and average responses to all three trial types (novel object, novel food, and habituation to a novel object) were significantly correlated. Although we saw no differences between neophobic and non-neophobic birds in IEG expression in response to novel objects in any of the 6 brain regions examined, there was a significant global decrease in ZENK expression and a significant increase in c-Fos expression in the medial ventral arcopallium and the caudal hippocampus in response to novel objects compared to controls, suggesting that these two regions may be important in novelty detection and threat perception. Additionally, there was no object effect in the rostral hippocampus, which supports the hypothesis that the avian hippocampus may have a rostrocaudal functional gradient similar to the septotemporal gradient in mammals.

Profiling of systemic and brain steroids in male songbirds: Seasonal changes in neurosteroids.

Steroids are secreted by the gonads and adrenal glands into the blood to modulate neurophysiology and behaviour. In addition, the brain can metabolise circulating steroids and synthesise steroids de novo. Songbirds show high levels of neurosteroid synthesis. In the present study, we developed and validated a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for the measurement of 10 steroids in whole blood, plasma and microdissected brain tissue (1-2 mg) of song sparrows. Our assay is highly accurate, precise, specific and sensitive. Moreover, the liquid-liquid extraction is fast, simple and effective. We quantified steroids in the blood and brain of wild male song sparrows in both breeding and non-breeding seasons. As expected, systemic androgen levels were higher in the breeding season than in the non-breeding season. Brain androgens were detectable only in the breeding season; androstenedione and 5α-dihydrotestosterone levels were up to 20-fold higher in specific brain regions than in blood. Oestrogens were not detectable in blood in both seasons. Oestrone and 17ß-oestradiol were detectable in brain in the breeding season only (up to 1.4 ng g-1 combined). Progesterone levels in several regions were higher in the non-breeding season than the breeding season, despite the lack of seasonal changes in systemic progesterone. Corticosterone levels in the blood were higher in the breeding season than in the non-breeding season but showed few seasonal differences in the brain. In general, the steroid levels presented here are lower than those in previous reports using immunoassays, because of the higher specificity of mass spectrometry. We conclude that (i) brain steroid levels can differ greatly from circulating steroid levels and (ii) brain steroid levels show region-specific seasonal patterns that are not a simple reflection of circulating steroid levels. This approach using ultrasensitive LC-MS/MS is broadly applicable to other species and allows steroid profiling in microdissected brain regions.

SLC2A12 of SLC2 Gene Family in Bird Provides Functional Compensation for the Loss of SLC2A4 Gene in Other Vertebrates.

Avian genomes are small and lack some genes that are conserved in the genomes of most other vertebrates including nonavian sauropsids. One hypothesis stated that paralogs may provide biochemical or physiological compensation for certain gene losses; however, no functional evidence has been reported to date. By integrating evolutionary analysis, physiological genomics, and experimental gene interference, we clearly demonstrate functional compensation for gene loss. A large-scale phylogenetic analysis of over 1,400 SLC2 gene sequences identifies six new SLC2 genes from nonmammalian vertebrates and divides the SLC2 gene family into four classes. Vertebrates retain class III SLC2 genes but partially lack the more recent duplicates of classes I and II. Birds appear to have completely lost the SLC2A4 gene that encodes an important insulin-sensitive GLUT in mammals. We found strong evidence for positive selection, indicating that the N-termini of SLC2A4 and SLC2A12 have undergone diversifying selection in birds and mammals, and there is a significant correlation between SLC2A12 functionality and basal metabolic rates in endotherms. Physiological genomics have uncovered that SLC2A12 expression and allelic variants are associated with insulin sensitivity and blood glucose levels in wild birds. Functional tests have indicated that SLC2A12 abrogation causes hyperglycemia, insulin resistance, and high relative activity, thus increasing energy expenditures that resemble a diabetic phenotype. These analyses suggest that the SLC2A12 gene not only functionally compensates insulin response for SLC2A4 loss but also affects daily physical behavior and basal metabolic rate during bird evolution, highlighting that older genes retain a higher level of functional diversification.

Time course of photo-induced Egr-1 expression in the hypothalamus of a seasonally breeding songbird.

Many seasonally-breeding species use daylength to time reproduction. Light-induced release of progonadal hormones involves a complex cascade of responses both inside and outside the brain. In this study, we used induction of early growth response 1 (Egr-1), the protein product of an immediate early gene, to evaluate the time course of such responses in male white-throated sparrows (Zonotrichia albicollis) exposed to a single long day. Induction of Egr-1 in the pars tuberalis began ∼11 h after dawn. This response was followed ∼6 h later by dramatic induction in the tuberal hypothalamus, including in the ependymal cells lining the third ventricle. At approximately the same time, Egr-1 was induced in dopaminergic and vasoactive intestinal peptide neurons in the tuberal hypothalamus and in dopaminergic neurons of the premammillary nucleus. We noted no induction in gonadotropin-releasing hormone (GnRH) neurons until 2 h after dawn the following morning. Overall, our results indicate that Egr-1 responses in GnRH neurons occur rather late during photostimulation, compared with responses in other cell populations, and that such induction may reflect new synthesis related to GnRH depletion rather than stimulation by light cues.

Long-term monitoring of mercury in adult saltmarsh sparrows breeding in Maine, Massachusetts and New York, USA 2000-2017.

Here we report on the results of a long-term study of mercury exposure in a songbird species, the saltmarsh sparrow (Ammodramus caudacutus). We measured total mercury concentrations in blood (n = 840) and feathers (n = 560) of adult saltmarsh sparrows at six locations between 2000 and 2017: Rachel Carson National Wildlife Refuge (RCNWR) in Wells, Maine; Scarborough Marsh State Wildlife Management Area in Scarborough, Maine; Parker River National Wildlife Refuge on Plum Island, Massachusetts; Pine Neck Preserve in Southampton, Long Island, New York; and North Cinder and North Green Sedge Islands off the coast of Long Island, New York. During the 12-17 year sampling periods, we found that mercury exposure differed by site and year but there was no consistent temporal trend across sites. Blood mercury concentrations declined only at RCNWR in Maine. We also found seasonal variation in blood mercury concentrations and a positive relationship between mercury concentrations of blood and innermost primary feather, but not between blood and tail feather.

Effects of stress-induced increases of corticosterone on circulating triglyceride levels, biliverdin concentration, and heme oxygenase expression.

When exposed to stressors, animals physiologically respond by secreting glucocorticoid hormones. Most birds, reptiles, and amphibians secrete corticosterone (CORT), which allows them to maximize short-term survival, including by modulating lipid metabolism. However, the factors regulating lipid metabolism, particularly during acute (i.e., short-term) stressors, are not well-characterized. To investigate one putative mechanism, we examined how expression of the enzyme heme oxygenase (HO), which primarily converts heme into biliverdin, changes during an acute stressor. Because HO has links to decreased levels of triglycerides, we tested the hypothesis that an acute stressor increases HO expression, which would concomitantly decrease circulating lipid levels. We compared free-living house sparrow (Passer domesticus) nestlings exposed to a one-hour stressor to control individuals, and quantified HO expression and biliverdin concentration in spleen, liver, or kidney tissue, as well as circulating CORT, triglyceride, and glycerol levels. Nestlings exposed to a stressor had reduced circulating triglycerides consistent with an increased rate of gluconeogenesis during an acute stressor. Concentrations of triglycerides were also negatively correlated with HO expression in the liver, which is consistent with mammalian studies. However, contrary to our predictions, exposure to a stressor did not affect HO expression, or biliverdin concentration in liver, spleen, or kidney. Overall, our results support links between CORT, triglyceride levels, and HO expression, though the molecular pathways connecting these metrics still need to be elucidated.

Molecular Diversification of the Seminal Fluid Proteome in a Recently Diverged Passerine Species Pair.

Seminal fluid proteins (SFPs) mediate an array of postmating reproductive processes that influence fertilization and fertility. As such, it is widely held that SFPs may contribute to postmating, prezygotic reproductive barriers between closely related taxa. We investigated seminal fluid (SF) diversification in a recently diverged passerine species pair (Passer domesticus and Passer hispaniolensis) using a combination of proteomic and comparative evolutionary genomic approaches. First, we characterized and compared the SF proteome of the two species, revealing consistencies with known aspects of SFP biology and function in other taxa, including the presence and diversification of proteins involved in immunity and sperm maturation. Second, using whole-genome resequencing data, we assessed patterns of genomic differentiation between house and Spanish sparrows. These analyses detected divergent selection on immunity-related SF genes and positive selective sweeps in regions containing a number of SF genes that also exhibited protein abundance diversification between species. Finally, we analyzed the molecular evolution of SFPs across 11 passerine species and found a significantly higher rate of positive selection in SFPs compared with the rest of the genome, as well as significant enrichments for functional pathways related to immunity in the set of positively selected SF genes. Our results suggest that selection on immunity pathways is an important determinant of passerine SF composition and evolution. Assessing the role of immunity genes in speciation in other recently diverged taxa should be prioritized given the potential role for immunity-related proteins in reproductive incompatibilities in Passer sparrows.

Slowing down the metabolic engine: impact of early-life corticosterone exposure on adult metabolism in house sparrows (Passer domesticus).

Whole-organism metabolism is an integrative process that determines not only the energy cost of living but also the energy output that is available for behavioral and physiological processes during the life cycle. Developmental challenge is known to affect growth, development of several organs, and several physiological mechanisms (such as HPA responsiveness, oxidative stress or immunity), which may altogether affect adult metabolism. All of these developmental effects are likely to be mediated by glucocorticoids, but the impact of developmental glucocorticoid exposure on adult metabolism has rarely been studied and the results are equivocal. In this study, we examined the impact of developmental exposure to corticosterone (CORT, the main avian glucocorticoid hormone) on resting metabolic rate (RMR, measured in thermoneutrality, 25°C) and thermoregulatory metabolic rate (TMR, measured in cold challenge conditions, 5°C) in the house sparrow. Following experimental administration of CORT at the nestling stage, house sparrows were kept in captivity until adulthood, when their metabolism was measured. We found that post-natal CORT exposure decreased both RMR and TMR in adult sparrows. This CORT-mediated reduction of metabolism was also associated with a reduced overnight body mass loss. Therefore, our results suggest that developmental CORT exposure can orient the phenotype towards an energy-saving strategy, which may be beneficial in a constraining environmental context.

Post-natal glucocorticoid elevation affects GnRH-induced luteinizing hormone concentration in female house sparrows.

Most non-mammalian studies investigating the long-term effects of early-life stressor exposure on endocrine regulation have focused on the hypothalamic-pituitaryadrenal (HPA) axis. However, the hypothalamic-pituitary-gonadal (HPG) axis may more directly affect fitness by regulating reproduction. Changes in HPG axis regulation could allow vertebrates to adaptively mitigate negative effects of early-life stressor exposure. However, only a few studies have examined long-term effects of early-life stressor experience on the HPG axis, and these have found mixed results. Here, we evaluate long-term effects of post-natal corticosterone exposure on the HPG axis in adult female house sparrows (Passer domesticus). We elevated circulating corticosterone non-invasively in wild nestling house sparrows between 8 and 11 days post-hatching, and then brought birds into captivity at fledging. Early in their first breeding season (ages 285-353d post-hatching), females were given a gonadotropin releasing hormone (GnRH) challenge. We found that early-life corticosterone exposure interacted with current condition such that females exposed to elevated post-natal corticosterone had higher baseline and GnRH-induced luteinizing hormone (LH) concentration than control females, but only if they had a high mass. Our results suggest that female house sparrows may mitigate negative impacts of early-life corticosterone exposure by investing in early reproduction, but only when current energetic condition allows.

Metal Exposure Risk Assessment for Tree Sparrows at Different Life Stages via Diet from a Polluted Area in Northwestern China.

To estimate the risk of metal (Cu, Zn, Pb, and Cd) exposure of tree sparrows through food at different life stages (including nestling, juvenile, adult in summer, and adult in winter), metal daily intake (MDI), metal daily intake per unit of body weight (MDIBW ), and exposure risks (hazard quotient and hazard index) of tree sparrows at different life stages from a polluted area, Baiyin, and a control area, Liujiaxia, were assessed. Metal daily intakes and MDIBW s of tree sparrows from Baiyin were higher than those from Liujiaxia, which can be attributed to higher metal concentrations of food sources in Baiyin than those in Liujiaxia. Metal daily intakes and MDIBW s of nestlings were higher than those of adults and juveniles. This difference is observed probably because nonphytophagous invertebrates, the predominant food fed to nestlings, possessed the highest metal concentrations. In addition, adults in summer had higher MDIs and MDIBW s than juveniles and adults in winter. This finding is probably observed because juveniles and adults in winter consumed a higher proportion of maize, which had the lowest metal concentrations. In addition, the biomagnification of 4 metals through the food chain increased the health risks of tree sparrows. Exposure risks of tree sparrows to metals were comparable to those of waterfowl and raptors, even when soil metals were below threshold. Environ Toxicol Chem 2019;38:2785-2796. © 2019 SETAC.

A multi-biomarker approach supports the use of compound-specific stable isotope analysis of amino acids to quantify basal carbon source use in a salt marsh consumer.

RATIONALE: Determining the flow of energy from primary producers to higher trophic levels in complex systems remains an important task for ecologists. Biomarkers can be used to trace carbon or energy sources contributing to an organism's tissues. However, different biomarkers vary in their ability to trace carbon sources based on how faithfully they transfer between trophic levels. Comparing emerging biomarker techniques with more commonly used techniques can demonstrate the relative efficacy of each in specific systems. METHODS: Two common biomarker techniques, fatty acid analysis (FAA) and bulk stable isotope analysis (SIA), and one emerging biomarker technique, compound-specific stable isotope analysis of amino acids (CSIA-AA), were compared to assess their ability to characterize and quantify basal carbon sources supporting the seaside sparrow (Ammodramus maritimus), a common salt marsh species. Herbivorous insect and deposit-feeding fiddler crab biomarker values were analyzed as proxies of major terrestrial and aquatic basal carbon sources, respectively. RESULTS: All three biomarker techniques indicated that both terrestrial and aquatic carbon sources were important to seaside sparrows. However, FAA could only be evaluated qualitatively, due to a currently limited understanding of trophic modification of fatty acids between primary producer and this consumer's tissues. Quantitative stable isotope (SIA or CSIA-AA) mixing models predicted nearly equal contributions of terrestrial and aquatic carbon sources supporting seaside sparrows, yet estimates based on CSIA-AA had greater precision. CONCLUSIONS: These findings support the use of CSIA-AA as an emerging tool to quantify the relative importance of basal carbon sources in salt marsh consumers. Integrating multiple biomarker techniques, with their differing benefits and limitations, will help to constrain models of carbon and energy flow in future ecosystem studies.

In vivo imaging of D2 receptors and corticosteroids predict behavioural responses to captivity stress in a wild bird.

Individual physiological variation may underlie individual differences in behaviour in response to stressors. This study tested the hypothesis that individual variation in dopamine and corticosteroid physiology in wild house sparrows (Passer domesticus, n = 15) would significantly predict behaviour and weight loss in response to a long-term stressor, captivity. We found that individuals that coped better with captivity (fewer anxiety-related behaviours, more time spent feeding, higher body mass) had lower baseline and higher stress-induced corticosteroid titres at capture. Birds with higher striatal D2 receptor binding (examined using positron emission tomography (PET) with 11C-raclopride 24 h post-capture) spent more time feeding in captivity, but weighed less, than birds with lower D2 receptor binding. In the subset of individuals imaged a second time, D2 receptor binding decreased in captivity in moulting birds, and larger D2 decreases were associated with increased anxiety behaviours 2 and 4 weeks post-capture. This suggests changes in dopaminergic systems could be one physiological mechanism underlying negative behavioural effects of chronic stress. Non-invasive technologies like PET have the potential to transform our understanding of links between individual variation in physiology and behaviour and elucidate which neuroendocrine phenotypes predict stress resilience, a question with important implications for both humans and wildlife.

Preparing to migrate: expression of androgen signaling molecules and insulin-like growth factor-1 in skeletal muscles of Gambel's white-crowned sparrows.

Migratory birds, including Gambel's white-crowned sparrows (Zonotrichia leucophrys gambelii), exhibit profound modifications of skeletal muscles prior to migration, notably hypertrophy of the pectoralis muscle required for powered flight. Muscle growth may be influenced by anabolic effects of androgens; however, prior to spring departure, circulating androgens are low in sparrows. A seasonal increase in local androgen signaling may occur within muscle to promote remodeling. We measured morphological parameters, plasma and tissue levels of testosterone, as well as mRNA expression levels of androgen receptor, 5α-reductase (converts testosterone to 5α-dihydrotestosterone), and the androgen-dependent myotrophic factor insulin-like growth factor-1. We studied the pectoralis muscle as well as the gastrocnemius (leg) muscle of male sparrows across three stages on the wintering grounds: winter (February), pre-nuptial molt (March), and pre-departure (April). Testosterone levels were low, but detectable, in plasma and muscles at all three stages. Androgen receptor mRNA and 5α-reductase Type 1 mRNA increased at pre-departure, but did so in both muscles. Notably, mRNA levels of insulin-like growth factor-1, an androgen-dependent gene critical for muscle remodeling, increased at pre-departure in the pectoralis but decreased in the gastrocnemius. Taken together, these data suggest a site-specific molecular basis for muscle remodeling that may serve to enable long-distance flight.

Daily expression of clock genes in central and peripheral tissues of tree sparrow (Passer montanus).

Almost all organisms live in a fluctuating environment. To achieve synchrony with the fluctuating environment, organisms have evolved with time-tracking mechanism commonly known as biological clocks. This circadian clock machinery has been identified in almost all cells of vertebrates and categorized as central and peripheral clocks. In birds, three independent circadian clocks reside within the nervous tissues in the hypothalamus, pineal and retina, which interact with each other and produce circadian time at a functional level. There is limited knowledge available of the molecular clockwork, and of integration between central and peripheral clocks in birds. Here, we studied daily expression of canonical clock genes (Bmal1, Clock, Per2, Per3, Cry1 and Cry2) and clock-controlled gene (Npas2) in all three central tissues (hypothalamus, pineal and retina) and in peripheral tissues (liver, intestine and muscle). Wild caught adult male tree sparrows were exposed to equinox photoperiod (12L:12D) for 2 weeks and after that birds were sacrificed (N = 5 per time point) at six time points (ZT1, ZT5, ZT9, ZT13, ZT17 and ZT21; ZT0 is lights on). Daily expression of clock genes was studied using qPCR. Bmal1, Clock, Per2, Per3, Cry1, Cry2 and Npas2 showed daily oscillation in all tissues except Cry2 in hypothalamus, pineal and intestine. We observed tissue-specific expression pattern for all clock and clock-controlled genes. Bmal1 transcripts expressed during early phase of night. Clock acrophase was observed during middle or late day time in the central clock while during early-to-middle phase of night in peripheral tissues. Npas2 expression pattern was similar to Bmal1. Per genes peaked either late at night or early during day time. However, Cry genes were peaked either at late day time (Cry1in retina, liver and intestine; Cry2 in liver and intestine) or at early night phase (Cry1 in hypothalamus, pineal and muscle; Cry2 in hypothalamus, pineal, retina and muscle). Our results are consistent with the autoregulatory circadian feedback loop, and suggest a conserved tissue-level circadian time generation in tree sparrows. Change in peak expression timing of these genes in different tissues implicates tissue-specific contribution of individual clock genes in the circadian time generation.