AbstractThe social environment is often the most dynamic and fitness-relevant environment animals experience. Here we tested whether plasticity arising from variation in social environments can promote signal-preference divergence-a key prediction of recent speciation theory but one that has proven difficult to test in natural systems. Interactions in mixed social aggregations could reduce, create, or enhance signal-preference differences. In the latter case, social plasticity could establish or increase assortative mating. We tested this by rearing two recently diverged species of Enchenopa treehoppers-sap-feeding insects that communicate with plant-borne vibrational signals-in treatments consisting of mixed-species versus own-species aggregations. Social experience with heterospecifics (in the mixed-species treatment) resulted in enhanced signal-preference species differences. For one of the two species, we tested but found no differences in the plastic response between sympatric and allopatric sites, suggesting the absence of reinforcement in the signals and preferences and their plastic response. Our results support the hypothesis that social plasticity can create or enhance signal-preference differences and that this might occur in the absence of long-term selection against hybridization on plastic responses themselves. Such social plasticity may facilitate rapid bursts of diversification.

Decision making resides at the interface between sensory perception and movement production. Female songbirds in the context of mate choice are an excellent system to define neural circuits through which sensory perception influences production of courtship behaviors. Previous experiments by our group and others have implicated secondary auditory brain sites, including the caudal nidopallium (NC), in mediating behavioral indicators of mate choice. Here, we used anterograde tracer molecules to define projections that emerge from NC in female songbirds, identifying pathways through which NC influences downstream sites implicated in signal processing and decision making. Our results reveal that NC sends projections into the arcopallium, including the ventral intermediate arcopallium (AIV). Previous work revealed that AIV also receives input from another auditory area implicated in song preference and mate choice (caudal mesopallium, CM), suggesting that convergent input from multiple auditory areas may play important roles in initiating mate choice behaviors. In the present results, NC projects to an area implicated in postural and locomotory control (dorsal arcopallium, Ad), suggesting that NC may play a role in directing those forms of copulatory behavior. NC projections also systematically avoid a vocal motor region of the arcopallium that is innervated by CM (robust nucleus of the arcopallium). These results suggest a model in which both NC and CM project to arcopallial pathways implicated in behavioral motivation. These brain regions may exert different influences on pathways through which auditory information can direct different facets of behavioral responses to information detected in those auditory signals.

Female songbirds use male song as an indicator of fitness and use that information to select their mate. Investigations of the female auditory system have provided evidence that the neurons within the caudal mesopallium (CM) are involved in the processing of songs that a female finds attractive, however, it is not clear how CM may exert its influence on behavioral indicators of mate choice. In the present study, anterograde tracing revealed the efferent connections of the female songbird CM. The results demonstrate connections to other auditory regions previously described in males, as well as novel connections to brain regions implicated in motor control. As in males, CM neurons in females project robustly to the lateral and medial extents of the caudal nidopallium, and to the ventral intermediate arcopallium. In a novel finding that is not present in males, CM neurons also project to the robust nucleus of the arcopallium and to the caudal striatum. Calling behavior and the expression of copulation solicitation displays are key indicators of female mate choice, and the projections found here bridge critical gaps necessary to understand how auditory perception can influence circuits related to the expression of those affiliative behaviors in female songbirds.

Mate selection can be stressful; time spent searching for mates can increase predation risk and/or decrease food consumption, resulting in elevated stress hormone levels. Both high predation risk and low food availability are often associated with increased variation in mate choice by females, but it is not clear whether stress hormone levels contribute to such variation in female behavior. We examined how the stress hormone corticosterone (CORT) affects female preferences for acoustic signals in the green treefrog, Hyla cinerea. Specifically, we assessed whether CORT administration affects female preferences for call rate - an acoustic feature that is typically under directional selection via mate choice by females in most anurans and other species that communicate using acoustic signals. Using a dual speaker playback paradigm, we show that females that were administered higher doses of CORT were less likely to choose male advertisement calls broadcast at high rates. Neither CORT dose nor level was related to the latency of female phonotactic responses, suggesting that elevated CORT does not influence the motivation to mate. Results were also not related to circulating sex steroids (i.e., progesterone, androgens or estradiol) that have traditionally been the focus of studies examining the hormonal basis for variation in female mate choice. Our results thus indicate that elevated CORT levels decrease the strength of female preferences for acoustic signals.

Male courtship displays may be regulated by, and affect the production of, circulating hormones. The Energetics-Hormone Vocalization (EHV) model, for example, posits that interactions among chorusing male anuran amphibians stimulate androgen production that then mediates an increase in vocal effort. Increased vocal effort is expected to deplete energy reserves and increase glucocorticoid levels that, in turn, negatively affect androgen levels and vocalization. Androgen levels, glucocorticoid levels, and vocal effort are thus expected to increase across and within nights of chorus activity and should be positively correlated in calling males; energy reserves should decline temporally and be inversely related to glucocorticoid levels. We tested predictions of the EHV model in the green treefrog, Hyla cinerea. Consistent with the model, both testosterone and dihydrotestosterone levels increased across the breeding season in calling males. However, testosterone levels decreased and dihydrotestosterone levels did not change within nights of chorus activity, suggesting that chorusing behavior did not drive the seasonal elevation in androgens. Corticosterone (CORT) level remained relatively stable across the breeding season and decreased within nights of chorus activity, contrary to model predictions. Body condition, the proxy for energetic state, was inversely correlated with CORT level but discrepancies between model predictions and temporal patterns of CORT production arose because there was no evidence of a temporal decrease in body condition or increase in vocal effort. Moreover, androgen and CORT levels were not positively correlated with vocal effort. Additional ecological and physiological measures may be needed to support predictions of the EHV model.