Females communicate non-genetic information about the environment to their developing offspring to generate potentially adaptive phenotypic variation. For example, birds allocate steroid hormones to their egg yolks in response to social stimuli and these hormones can induce long-lasting changes to offspring physiology and behavior. However, little work has explored how multiple yolk steroids respond to social challenges, despite their shared biosynthetic pathway, or how they impact early development. Here, we conducted simulated territorial intrusions to identify socially responsive yolk steroids using liquid chromatography with tandem mass spectroscopy (LC-MS-MS) and explored their relationship with gene expression patterns in extra-embryonic membranes during the earliest stages of embryonic development in house sparrows (Passer domesticus). We also characterized how yolk hormones changed across development, from embryonic day 0 (ED0) to ED5. We found that yolk hormone concentrations largely declined with embryonic development, but at ED0 multiple hormones along the androgenic pathway were elevated in response to the number of territorial intrusions a female experienced, with yolk testosterone and progesterone showing the strongest responses. At ED3, these socially responsive yolk hormones were related to gene expression in extra-embryonic membranes linked to key developmental processes, such as growth and immune function. These data highlight a critical early window during which yolk hormones may impact offspring traits through changes in extra-embryonic gene expression.

Uncovering the genomic bases of phenotypic adaptation is a major goal in biology, but this has been hard to achieve for complex behavioural traits. Here we leverage the repeated, independent evolution of obligate cavity nesting in birds to test the hypothesis that pressure to compete for a limited breeding resource has facilitated convergent evolution in behaviour, hormones and gene expression. We used an integrative approach, combining aggression assays in the field, testosterone measures and transcriptome-wide analyses of the brain in wild-captured females and males. Our experimental design compared species pairs across five avian families, each including one obligate cavity-nesting species and a related species with a more flexible nest strategy. We find behavioural convergence, with higher levels of territorial aggression in obligate cavity nesters, particularly among females. Across species, levels of testosterone in circulation were not associated with nest strategy nor aggression. Phylogenetic analyses of individual genes and co-regulated gene networks revealed more shared patterns of brain gene expression than expected by drift, although the scope of convergent gene expression evolution was limited to a small percentage of the genome. When comparing our results to other studies that did not use phylogenetic methods, we suggest that accounting for shared evolutionary history may reduce the number of genes inferred as convergently evolving. Altogether, we find that behavioural convergence in response to shared ecological pressures is associated with largely independent evolution of gene expression across different avian families, punctuated by a narrow set of convergently evolving genes.

Offspring from females breeding in competitive social environments are often exposed to more testosterone (T) during embryonic development, which can affect traits from growth to behavior in potentially adaptive ways. Despite the important role of maternally derived steroids in shaping offspring development, the molecular mechanisms driving these processes are currently unclear. Here, we use tree swallows (Tachycineta bicolor) to explore the effects of the maternal social environment on yolk T concentrations and genome-wide patterns of neural gene expression in embryos. We measured aggressive interactions among females breeding at variable densities and collected their eggs at two timepoints, including the day laid to measure yolk T concentrations and on embryonic day 11 to measure gene expression in whole brain samples. We found that females breeding in high-density sites experienced elevated rates of physical aggression and their eggs had higher yolk T concentrations. A differential gene expression and weighted gene co-expression network analysis indicated that embryos from high-density sites experienced an upregulation of genes involved in hormone, circulatory, and immune processes, and these gene expression patterns were correlated with yolk T levels and aggression. Genes implicated in neural development were additionally downregulated in embryos from high-density sites. These data highlight how early neurogenomic processes may be affected by the maternal social environment, giving rise to phenotypic plasticity in offspring.

The prenatal transfer of testosterone (T) from mother to offspring is an important source of phenotypic plasticity. In birds, exposure to ecologically relevant stimuli, such as social competition or an attractive mate, can cause females to deposit more T into their egg yolks. Exposure to elevated yolk T can modify the expression of several fitness-related traits in offspring (e.g. growth, immune function, secondary sex traits and behaviour). Despite some of these changes being potentially adaptive, not all studies find that yolk T levels change in response to ecologically relevant stimuli. This heterogeneity is currently unexplained, limiting our ability to predict inter-generational responses to ecological change. Here, we performed a systematic literature search and found 119 observations across 39 wild species that measured inter-female variation in yolk T allocation in response to various stimuli. We used boosted regression trees, a form of machine learning, to examine whether species-specific traits or variation in study-level variables could explain variation in yolk T allocation (i.e. statistically significant vs. non-significant responses). We found that both species-specific traits and study-level variables are important predictors of significant changes in yolk T levels. Geographic range (latitude and longitude), evolutionary distinctiveness, longevity, egg mass relative to female mass, sociality, migration status and time to fledge were among the top 10 most influential predictors of the 48 examined. We also found that studies measuring or manipulating social stimuli (e.g. competition and breeding density) or breeding date were more likely to detect changes in yolk T allocation compared with studies examining other ecological contexts. Overall, these data provide several testable hypotheses concerning yolk T allocation and its adaptive value across species and contexts. Additionally, these findings can help us predict how ecological changes will affect hormonal responses in females that can shape future generations. Among birds, yolk testosterone allocation patterns are not consistent across species or studies, which dampens our ability to understand what drives and shapes maternal effects. Here, the authors examine ecological factors and species-specific traits that may shape these responses and generate hypotheses for investigating differences in yolk T allocation.image

Ecogeographic rules describe spatial patterns in biological trait variation and shed light on the drivers of such variation. In animals, a consensus is emerging that 'pioneering' traits may facilitate range shifts via a set of bold, aggressive and stress-resilient traits. Many of these same traits are associated with more northern latitudes, and most range shifts in the northern hemisphere indicate northward movement. As a consequence, it is unclear whether pioneering traits are simply corollaries of existing latitudinal variation, or whether they override other well-trodden latitudinal patterning as a unique ecogeographic rule of phenotypic variation. The tree swallow Tachycineta bicolor is a songbird undergoing a southward range shift in the eastern United States, in direct opposition of the poleward movement seen in most other native species' range shifts. Because this organic range shift countervails the typical direction of movement, this case study provides for unique ecological insights on organisms and their ability to thrive in our changing world. We sampled female birds across seven populations, quantifying behavioural, physiological and morphological traits. We also used GIS and field data to quantify a core set of ecological factors with strong ties to these traits as well as female performance. Females at more southern expansion sites displayed higher maternal aggression, higher baseline corticosterone and more pronounced elevation of corticosterone following a standardized stressor, contrary to otherwise largely conserved latitudinal patterning in these traits. Microhabitat variation explained some quantitative phenotypic variation, but the expansion and historic ranges did not differ in openness, distance to water or breeding density. This countervailing range shift therefore suggests that pioneering traits are not simply corollaries of existing latitudinal variation, but rather, they may override other well-trodden latitudinal patterning as a unique ecogeographic rule of phenotypic variation.

Studies of the evolutionary causes and consequences of variation in circulating glucocorticoids (GCs) have begun to reveal how they are shaped by selection. Yet the extent to which variation in circulating hormones reflects variation in other important regulators of the hypothalamic-pituitary-adrenal (HPA) axis, and whether these relationships vary among populations inhabiting different environments, remain poorly studied. Here, we compare gene expression in the brain of female tree swallows (Tachycineta bicolor) from populations that breed in environments that differ in their unpredictability. We find evidence of inter-population variation in the expression of glucocorticoid and mineralocorticoid receptors in the hypothalamus, with the highest gene expression in a population from an extreme environment, and lower expression in a population from a more consistent environment as well as in birds breeding at an environmentally variable high-altitude site that are part of a population that inhabits a mixture of high and low altitude habitats. Within some populations, variation in circulating GCs predicted differences in gene expression, particularly in the hypothalamus. However, some patterns were present in all populations, whereas others were not. These results are consistent with the idea that some combination of local adaptation and phenotypic plasticity may modify components of the HPA axis affecting stress resilience. Our results also underscore that a comprehensive understanding of the function and evolution of the stress response cannot be gained from measuring circulating hormones alone, and that future studies that apply a more explicitly evolutionary approach to important regulatory traits are likely to provide significant insights.

The ovary plays an important role in mediating both a female's response to her social environment and communicating it to her developing offspring via maternal effects. Past work has focused on how ovarian hormones respond to competition, but we know little about how the broader ovarian transcriptomic landscape changes, either during or after competition, giving us a narrow perspective on how socially induced phenotypes arise. Here, we experimentally generated social competition among wild, cavity-nesting female birds (tree swallows, Tachycineta bicolor), a species in which females lack a socially induced rise in circulating testosterone but they nevertheless increase allocation to eggs. After territory settlement, we reduced availability of nesting cavities, generating heightened competition; within 24 h we reversed the manipulation, causing aggressive interactions to subside. We measured ovarian transcriptomic responses at the peak of competition and 48 h later, along with date-matched controls. Network analyses indicated that competing females experienced an immediate and temporary decrease in the expression of genes involved in the early stages of steroidogenesis, and this was moderately correlated with plasma testosterone; however, two days after competition had ended, there was a marked increase in the expression of genes involved in the final stages of steroidogenesis, including HSD17B1. Gene networks related to the cell cycle, muscle performance, and extracellular matrix organization also displayed altered activity. Although the functional consequences of these findings are unclear, they shed light on socially responsive ovarian genomic mechanisms that could potentially exert lasting effects on behavior, reproduction, and maternal effects.

Periods of social instability can elicit adaptive phenotypic plasticity to promote success in future competition. However, the underlying molecular mechanisms have primarily been studied in captive and laboratory-reared animals, leaving uncertainty as to how natural competition among free-living animals affects gene activity. Here, we experimentally generated social competition among wild, cavity-nesting female birds (tree swallows, Tachycineta bicolor). After territorial settlement, we reduced the availability of key breeding resources (i.e., nest boxes), generating heightened competition; within 24 h we reversed the manipulation, causing aggressive interactions to subside. We sampled females during the peak of competition and 48 h after it ended, along with date-matched controls. We measured transcriptomic and epigenomic responses to competition in two socially relevant brain regions (hypothalamus and ventromedial telencephalon). Gene network analyses suggest that processes related to energy mobilization and aggression (e.g., dopamine synthesis) were up-regulated during competition, the latter of which persisted 2 d after competition had ended. Cellular maintenance processes were also down-regulated after competition. Competition additionally altered methylation patterns, particularly in pathways related to hormonal signaling, suggesting those genes were transcriptionally poised to respond to future competition. Thus, experimental competition among free-living animals shifts gene expression in ways that may facilitate the demands of competition at the expense of self-maintenance. Further, some of these effects persisted after competition ended, demonstrating the potential for epigenetic biological embedding of the social environment in ways that may prime individuals for success in future social instability.

Timberlake was, unquestionably, an ethologist’s psychologist--his cross-disciplinary incorporation of biology defining his research program. Centering his notion of “biological behaviorism,” I provide a brief overview of the life and work of William Timberlake. Timberlake, like other behaviorism-influenced researchers, saw value in a detailed understanding of the behavioral structure of organisms without necessarily exploring just how, causally, these behaviors are mediated. This framing shaped his work from his early training in Hullian behaviorism, to his initial flirtations with ethology, to his pioneering disequilibrium account of reinforcement, and ultimately to his trademark behavior systems approach. Bringing in new insights about the animal and its ecology allowed for Timberlake’s work on learning to achieve marked predictive success. However, there was more to his project, and I conclude by placing Timberlake’s animal-centered biological behaviorism in the context of his integrative vision for the science of animal behavior (and for CISAB).

Decades of work indicate that female birds can control their offspring sex ratios in response to environmental and social cues. In laying hens, hormones administered immediately prior to sex chromosome segregation can exert sex ratio skews, indicating that these hormones may act directly on the germinal disc to influence which sex chromosome is retained in the oocyte and which is discarded into an unfertilizable polar body. We aimed to uncover the gene pathways involved in this process by testing whether treatments with testosterone or corticosterone that were previously shown to influence sex ratios elicit changes in the expression of genes and/or gene pathways involved in the process of meiotic segregation. We injected laying hens with testosterone, corticosterone, or control oil 5h prior to ovulation and collected germinal discs from the F1 preovulatory follicle in each hen 1.5h after injection. We used RNA-sequencing (RNA-seq) followed by DESeq2 and gene set enrichment analyses to identify genes and gene pathways that were differentially expressed between germinal discs of control and hormone-treated hens. Corticosterone treatment triggered downregulation of 13 individual genes, as well as enrichment of gene sets related to meiotic spindle organization and chromosome segregation, and additional gene sets that function in ion transport. Testosterone treatment triggered upregulation of one gene, and enrichment of one gene set that functions in nuclear chromosome segregation. This work indicates that corticosterone can be a potent regulator of meiotic processes and provides potential gene targets on which corticosterone and/or testosterone may act to influence offspring sex ratios in birds.

Competitive interactions often occur in series; therefore animals may respond to social challenges in ways that prepare them for success in future conflict. Changes in the production of the steroid hormone testosterone (T) are thought to mediate phenotypic responses to competition, but research over the past few decades has yielded mixed results, leading to several potential explanations as to why T does not always elevate following a social challenge. Here, we measured T levels in tree swallows (Tachycineta bicolor), a system in which females compete for limited nesting cavities and female aggression is at least partially mediated by T. We experimentally induced social challenges in two ways: (1) using decoys to simulate territorial intrusions and (2) removing subsets of nesting cavities to increase competition among displaced and territory-holding females. Critically, these experiments occurred pre-laying, when females are physiologically capable of rapidly increasing circulating T levels. However, despite marked aggression in both experiments, T did not elevate following real or simulated social challenges, and in some cases, socially challenged females had lower T levels than controls. Likewise, the degree of aggression was negatively correlated with T levels following a simulated territorial intrusion. Though not in line with the idea that social challenges prompt T elevation in preparation for future challenges, these patterns nevertheless connect T to territorial aggression in females. Coupled with past work showing that T promotes aggression, these results suggest that T may act rapidly to allow animals to adaptively respond to the urgent demands of a competitive event.