Mate Preferences and Choosiness Are Distinct Components of Mate Choice in Eastern Gray Treefrogs (Hyla versicolor).

AbstractMate choice is an important cause of natural and sexual selection, driving the evolution of ornaments and promoting diversification and speciation. Mate choice decisions arise from the interaction of several components, and knowledge of whether they interact, and how, is crucial for understanding their contributions to selection. Here we focus on the relationship between preference functions (attractiveness ranking of prospective mates) and choosiness (effort invested in obtaining the preferred mate) and test the hypothesis that they are independent components of mate choice decisions. We examine individual variation in preference functions and choosiness for call duration in female Hyla versicolor treefrogs and show that measures describing preference functions and choosiness are not correlated. We also found a suggestive but inconclusive pattern that both components are influenced by different factors (body measures and hormones). Independence of preference and choosiness suggests that the joint study of variation in both components is required to gain a complete understanding of how mate choice contributes to sexual selection and speciation.

Positive-to-negative behavioural responses suggest hedonic evaluation in treefrog mate choice.

Sexual competition hinges on the ability to impress other conspecifics, to drive them away or attract them. In such cases, the selective environment may be hedonic or affective in nature, as it consists of the evaluations of the individuals making the decisions. This may contribute to the power of sexual selection because evaluations may range from positive to negative rather than simply from positive to neutral. Selection due to mate choice may therefore be stronger than currently appreciated. Further, change in preferred mate types can occur simply by changes (flips) in the evaluation of similar display features, adding to the dynamism of sexual selection as well as its strength. We tested the hypothesis of positive-to-negative behavioural responses in mate choice with a playback experiment using two treefrog species with 'mirror image' structures in their advertisement and aggressive calls. Female treefrog responses ranged from approach to evasion, and the presence of an aversive stimulus tainted evaluation of an attractive stimulus. Further, females in the two species showed flips in approach/evasion of stimuli with comparable signal structure. These results suggest that hedonic evaluation may have an important role in mate choice and showcase how mechanistic analysis can help understand evolutionary processes.

On the architecture of mate choice decisions: preference functions and choosiness are distinct traits.

Mate choice is an important cause of sexual selection; it can drive the evolution of extravagant ornaments and displays, and promote speciation through the reproductive isolation generated by rapid divergence of sexual traits. Understanding mate choice requires knowledge of the traits involved in generating mate-choice decisions, and how those traits may interact with each other. It has been proposed that mate-choice decisions are the outcome of two components that vary independently: the preference function (the ranking of the attractiveness of prospective mates) and choosiness (the effort invested in mate assessment). Here we test this hypothesis by examining individual variation in female preference functions and choosiness in green treefrogs ( Hyla cinerea). We show that measures describing preference functions and choosiness are not correlated. We also show that both components are influenced differently by variation in female body size, and that preference function shape (closed and preferring intermediate values or open-ended and preferring extremes) has a strong influence on this relationship: function traits are positively correlated with body size only for individuals with closed functions, while choosiness is positively correlated with body size for individuals with open functions, but negatively for those with closed functions.

Frequency channel-dependent selectivity for temporal call characteristics in gray treefrogs, Hyla versicolor.

Sensory receptors transmit information on multiple stimulus dimensions. Much remains to be understood about how the processing of different signal characteristics is partitioned and integrated in different areas of the nervous system. Amphibian hearing involves two morphologically distinct inner-ear organs that process different components of the frequency spectrum. Many anuran signals contain two frequency peaks, each one matching the sensitivity of one of these two organs. We hypothesized that the processing of temporal characteristics of acoustic signals would differ in these two frequency channels, perhaps because of differences in the response properties of the two inner-ear organs. We tested this hypothesis in the gray treefrog, Hyla versicolor; male advertisement calls of this species contain a bimodal frequency spectrum. We generated synthetic male advertisement calls in which we independently manipulated the pattern of amplitude modulation in the low-frequency peak or the high-frequency peak and measured the attractiveness of these stimuli to females in single-speaker and two-speaker phonotaxis tests. We obtained multiple lines of evidence that females were more selective for fine-temporal characteristics in the high-frequency peak. We discuss the potential implications of frequency channel-dependent temporal processing for signal evolution and suggest that additional neurophysiological investigations of the anuran auditory periphery will give important insights into how the nervous system partitions the encoding of multiple characteristics of complex signals.

Nephila clavipes spiders (Araneae: Nephilidae) keep track of captured prey counts: testing for a sense of numerosity in an orb-weaver.

Nephila clavipes golden orb-web spiders accumulate prey larders on their webs and search for them if they are removed from their web. Spiders that lose larger larders (i.e., spiders that lose larders consisting of more prey items) search for longer intervals, indicating that the spiders form memories of the size of the prey larders they have accumulated, and use those memories to regulate recovery efforts when the larders are pilfered. Here, we ask whether the spiders represent prey counts (i.e., numerosity) or a continuous integration of prey quantity (mass) in their memories. We manipulated larder sizes in treatments that varied in either prey size or prey numbers but were equivalent in total prey quantity (mass). We then removed the larders to elicit searching and used the spiders' searching behavior as an assay of their representations in memory. Searching increased with prey quantity (larder size) and did so more steeply with higher prey counts than with single prey of larger sizes. Thus, Nephila spiders seem to track prey quantity in two ways, but to attend more to prey numerosity. We discuss alternatives for continuous accumulator mechanisms that remain to be tested against the numerosity hypothesis, and the evolutionary and adaptive significance of evidence suggestive of numerosity in a sit-and-wait invertebrate predator.

Female gray treefrogs, Hyla versicolor, are responsive to visual stimuli but unselective of stimulus characteristics.

The visual ecology of nocturnal anurans is poorly understood, but there is growing evidence that vision plays a role in important behaviors such as mate choice. While several recent studies have demonstrated that females are responsive to visual cues when selecting mates, the forces responsible for these preferences are unknown. We investigated the responsiveness of female gray treefrogs, Hyla versicolor, to video playbacks of calling conspecific males in which we varied attributes of the vocal sac, a conspicuous visual characteristic of calling males and a target of female choice in other species. Females responded surprisingly strongly to the video playbacks, but did so indiscriminately with respect to variation in vocal sac characteristics. We followed up on these results with a series of additional tests that examined female responses to abstract stimuli. Females continued to respond to such stimuli, leading us to conclude that their behavior was related to a generalized phototactic response. Because of this, we were unable to make conclusions regarding female preferences for vocal sac characteristics. Nonetheless, our results are significant in two respects. First, we illustrate that despite much effort into improving video playback methodologies, challenges remain, and we offer our experimental design as a method to ensure that appropriate conclusions can be drawn from such studies. Second, we argue that the female phototactic response has potentially significant behavioral implications and in general the consequences of anuran visual preferences deserve further investigation.

Diversification under sexual selection: the relative roles of mate preference strength and the degree of divergence in mate preferences.

The contribution of sexual selection to diversification remains poorly understood after decades of research. This may be in part because studies have focused predominantly on the strength of sexual selection, which offers an incomplete view of selection regimes. By contrast, students of natural selection focus on environmental differences that help compare selection regimes across populations. To ask how this disparity in focus may affect the conclusions of evolutionary research, we relate the amount of diversification in mating displays to quantitative descriptions of the strength and the amount of divergence in mate preferences across a diverse set of case studies of mate choice. We find that display diversification is better explained by preference divergence rather than preference strength; the effect of the latter is more subtle, and is best revealed as an interaction with the former. Our findings cast the action of sexual selection (and selection in general) in a novel light: the strength of selection influences the rate of evolution, and how divergent selection is determines how much diversification can occur. Adopting this view will enhance tests of the relative role of natural and sexual selection in processes such as speciation.

Dip listening or modulation masking? Call recognition by green treefrogs (Hyla cinerea) in temporally fluctuating noise.

Despite the importance of perceptually separating signals from background noise, we still know little about how nonhuman animals solve this problem. Dip listening, an ability to catch meaningful 'acoustic glimpses' of a target signal when fluctuating background noise levels momentarily drop, constitutes one possible solution. Amplitude-modulated noises, however, can sometimes impair signal recognition through a process known as modulation masking. We asked whether fluctuating noise simulating a breeding chorus affects the ability of female green treefrogs (Hyla cinerea) to recognize male advertisement calls. Our analysis of recordings of the sounds of green treefrog choruses reveal that their levels fluctuate primarily at rates below 10 Hz. In laboratory phonotaxis tests, we found no evidence for dip listening or modulation masking. Mean signal recognition thresholds in the presence of fluctuating chorus-like noises were never statistically different from those in the presence of a non-fluctuating control. An analysis of statistical effects sizes indicates that masker fluctuation rates, and the presence versus absence of fluctuations, had negligible effects on subject behavior. Together, our results suggest that females listening in natural settings should receive no benefits, nor experience any additional constraints, as a result of level fluctuations in the soundscape of green treefrog choruses.

Vibrational playback by means of airborne stimuli.

Substrate-borne vibrational communication is a common form of communication in animals. Current contact-based playback methods limit the number of substrates that can be stimulated simultaneously and potentially change the transmission properties of the substrate. Here, we explore a solution to these limitations by broadcasting airborne stimuli onto plant substrates to impart vibrational playbacks. We demonstrate that one can effectively compensate for the filtering properties of any individual plant across a range of frequencies. We then address how well both compensated broad-band and pure-tone stimuli for one plant individual apply to other individuals across days. Variation within and between plants was similar across the range tested but was quite variable at certain frequencies. Focusing on a subset of this range, at low frequencies, responses were flat across days and pure-tone frequency stimuli in this range were consistently transmitted despite repositioning of plants relative to the loudspeaker. Our results present a potential solution to researchers interested in exposing large samples of individuals to vibrational signals but also highlight the importance of validating the use of airborne stimuli as vibrational playbacks to the particular substrate type and frequency range of interest.

Woe is the loner: Female treefrogs prefer clusters of displaying males over single "hotshot" males.

Communal displays such as leks and choruses are puzzling phenomena, as it is not obvious why signalers or choosers should aggregate. It has been hypothesized that signalers in leks enjoy higher per-capita reproductive success because choosers prefer to sample among dense configurations ("clusters") that are easier to compare. Although female preferences as well as the signal features of attractive males are well characterized in many chorusing species, we know little about how mate sampling is influenced by the spatial dynamics within communal displays. Here, we ask how female Eastern Gray Treefrogs (Hyla versicolor) respond to isolated and clustered call stimuli in a simple one versus three playback design. We explored (i) whether females exhibit a general preference for call clusters, (ii) whether spatial preference is robust to call-feature preference, and (iii) how this affects the relative success of attractive and unattractive males in different spatial combinations. We found generalized spatial discrimination against lone callers but did observe fine-scale assessment of call features within clusters. The prominence of the spatial preference impacts the attractiveness of males, conferring particular advantage to attractive callers within clusters, while reducing attractiveness of isolated males regardless of their acoustic features. Our findings indicate that female frogs navigate complex choruses by initially orientating toward clusters of calling males, and then assess call features within them. This study provides novel insight into the mate choice heuristics involved in animal choruses.

Phenotypic integration and the evolution of signal repertoires: A case study of treefrog acoustic communication.

Animal signals are inherently complex phenotypes with many interacting parts combining to elicit responses from receivers. The pattern of interrelationships between signal components reflects the extent to which each component is expressed, and responds to selection, either in concert with or independently of others. Furthermore, many species have complex repertoires consisting of multiple signal types used in different contexts, and common morphological and physiological constraints may result in interrelationships extending across the multiple signals in species' repertoires. The evolutionary significance of interrelationships between signal traits can be explored within the framework of phenotypic integration, which offers a suite of quantitative techniques to characterize complex phenotypes. In particular, these techniques allow for the assessment of modularity and integration, which describe, respectively, the extent to which sets of traits covary either independently or jointly. Although signal and repertoire complexity are thought to be major drivers of diversification and social evolution, few studies have explicitly measured the phenotypic integration of signals to investigate the evolution of diverse communication systems. We applied methods from phenotypic integration studies to quantify integration in the two primary vocalization types (advertisement and aggressive calls) in the treefrogs Hyla versicolor, Hyla cinerea, and Dendropsophus ebraccatus. We recorded male calls and calculated standardized phenotypic variance-covariance (P) matrices for characteristics within and across call types. We found significant integration across call types, but the strength of integration varied by species and corresponded with the acoustic similarity of the call types within each species. H. versicolor had the most modular advertisement and aggressive calls and the least acoustically similar call types. Additionally, P was robust to changing social competition levels in H. versicolor. Our findings suggest new directions in animal communication research in which the complex relationships among the traits of multiple signals are a key consideration for understanding signal evolution.

Beyond temperature coupling: Effects of temperature on ectotherm signaling and mate choice and the implications for communication in multispecies assemblages.

Many organisms share communication channels, generating complex signaling environments that increase the risk of signal interference. Variation in abiotic conditions, such as temperature, may further exacerbate signal interference, particularly in ectotherms. We tested the effects of temperature on the pulse rate of male signals in a community of Oecanthus tree crickets, and for one focal species we also assessed its effect on female pulse rate preferences and motivation to seek mates. We confirm prior findings of temperature-dependent signals that result in increasing signal similarity at lower temperatures. Temperature also affected several aspects of female preferences: The preferred pulse rate value was temperature dependent, and nearly perfectly coupled with signal pulse rate; the range of pulse rate values that females found attractive also increased with temperature. By contrast, the motivation of females to perform phonotaxis was unaffected by temperature. Thus, at lower temperatures the signals of closely related species were more similar and females more discriminating. However, because signal similarity increased more strongly than female discrimination, signal interference and the likelihood of mismating may increase as temperatures drop. We suggest that a community approach will be useful for understanding the role of environmental variability in the evolution of communication systems.

Evolution of advertisement signals in North American hylid frogs: vocalizations as end-products of calling behavior.

We studied the advertisement signals in two clades of North American hylid frogs in order to characterize the relationships between signal acoustic structure and underlying behavior. A mismatch was found between the acoustic structure and the mechanism of sound production. Two separate sets of phylogenetic characters were coded following acoustic versus mechanistic criteria, and exploratory treatments were made to compare their respective phylogenetic content in comparison with the molecular phylogeny (Faivovich et al., 2005). We discuss the consequences of the acoustic/mechanistic mismatch in terms of significance of acoustic characters for phylogenetic and comparative studies; and the evolution of vocalizations in North American treefrogs. Considering only the acoustic structure of frog vocalizations can lead to misleading results in terms of both phylogenetic signal and evolution of vocalizations. In contrast, interpreting the acoustic signals with regard to the mechanism of sound production results in consistent phylogenetic information. The mechanistic coding also provides strong homologies for use in comparative studies of frog vocalizations, and to derive and test evolutionary hypotheses.

Do Green Treefrogs Use Social Information to Orient Outside the Breeding Season?

Gerlinde Höbel and Ashley Christie (2016) To decide efficiently where to forage, rest or breed, animals need information about their environment, which they may gather by monitoring the behavior of others. For example, attending to the signals of conspecifics or heterospecifics with similar habitat requirements may facilitate habitat choice. Such social information use seems taxonomically widespread, yet there is currently a dearth of information for amphibians. Anuran amphibians, with their highly developed auditory system and robust phonotaxis towards advertisement calls when searching for mates seem predisposed to use this hearing capability in other behavioral contexts. We conducted playback experiments to test whether anurans exploit acoustic signals in a non-reproductive context. In our experiments female Green Treefrogs did not show phonotaxis to signals associated with the presence of other frogs, and the orientation and speed of their movement was not different from animals randomly moving inside a silent arena. Previous studies documenting social information use in anurans have tested reproductively active frogs during the breeding season. By contrast, our study examined non-reproductive animals, and these did not approach social signals. We propose two non-exclusive hypotheses for this observed difference in phonotaxis behavior: (1) attending to social signals is restricted to ecologically most relevant time periods in a frogs life (i.e., finding breeding sites during the mating season), or (2) the ability of acoustic signals to stimulate the auditory system may be influenced by hormone levels regulating the reproductive state.

Modality interactions alter the shape of acoustic mate preference functions in gray treefrogs.

Sexual selection takes place in complex environments where females evaluating male mating signals are confronted with stimuli from multiple sources and modalities. The pattern of expression of female preferences may be influenced by interactions between modalities, changing the shape of female preference functions, and thus ultimately altering the selective landscape acting on male signal evolution. We tested the hypothesis that the responses of female gray treefrogs, Hyla versicolor, to acoustic male advertisement calls are affected by interactions with visual stimuli. We measured preference functions for several call traits under two experimental conditions: unimodal (only acoustic signals presented), and multimodal (acoustic signals presented along with a video-animated calling male). We found that females were more responsive to multimodal stimulus presentations and, compared to unimodal playbacks, had weaker preferences for temporal call characteristics. We compared the preference functions obtained in these two treatments to the distribution of male call characteristics to make inferences on the strength and direction of selection expected to act on male calls. Modality interactions have the potential to influence the course of signal evolution and thus are an important consideration in sexual selection studies.

Reproductive character displacement in the acoustic communication system of green tree frogs (Hyla cinerea).

Interactions between species can affect the evolution of their sexual signals, receiver selectivity, or both. One commonly expected outcome is reproductive character displacement, whereby adverse consequences of mismating select for greater differentiation of communication systems in areas of sympatry than in areas of allopatry. We found evidence of reproductive character displacement in the acoustic communication system of green tree frogs (Hyla cinerea). The strength of female preferences for the spectral properties of calls that distinguish conspecific calls from those of a closely related congener, H. gratiosa, was greater in areas of sympatry with H. gratiosa than in areas of allopatry. We also found subtle differences in advertisement calls and in the heights of male calling perches when we restricted our comparisons to localities in which H. gratiosa was also breeding (syntopy) with localities where this species was absent. Hyla cinerea and H. gratiosa show only weak genetic incompatibility, but the calls representative of interspecific hybrids were unattractive to females of both parental species. Hybrids might also be at an ecological disadvantages because of different habitat preferences of the two taxa. Thus, selection against production of less fit or less attractive hybrid or backcross offspring are probably the main causes responsible for the differences documented in this paper.

Effect of temporal and spectral noise features on gap detection behavior by calling green treefrogs.

Communication plays a central role in the behavioral ecology of many animals, yet the background noise generated by large breeding aggregations may impair effective communication. A common behavioral strategy to ameliorate noise interference is gap detection, where signalers display primarily during lulls in the background noise. When attempting gap detection, signalers have to deal with the fact that the spacing and duration of silent gaps is often unpredictable, and that noise varies in its spectral composition and may thus vary in the degree in which it impacts communication. I conducted playback experiments to examine how male treefrogs deal with the problem that refraining from calling while waiting for a gap to appear limits a male's ability to attract females, yet producing calls during noise also interferes with effective sexual communication. I found that the temporal structure of noise (i.e., duration of noise and silent gap segments) had a stronger effect on male calling behavior than the spectral composition. Males placed calls predominantly during silent gaps and avoided call production during short, but not long, noise segments. This suggests that male treefrogs use a calling strategy that maximizes the production of calls without interference, yet allows for calling to persist if lulls in the background noise are infrequent.

Listening for males and bats: spectral processing in the hearing organ of Neoconocephalus bivocatus (Orthoptera: Tettigoniidae).

Tettigoniids use hearing for mate finding and the avoidance of predators (mainly bats). Using intracellular recordings, we studied the response properties of auditory receptor cells of Neoconocephalus bivocatus to different sound frequencies, with a special focus on the frequency ranges representative of male calls and bat cries. We found several response properties that may represent adaptations for hearing in both contexts. Receptor cells with characteristic frequencies close to the dominant frequency of the communication signal were more broadly tuned, thus extending their range of high sensitivity. This increases the number of cells responding to the dominant frequency of the male call at low signal amplitudes, which should improve long distance call localization. Many cells tuned to audio frequencies had intermediate thresholds for ultrasound. As a consequence, a large number of receptors should be recruited at intermediate amplitudes of bat cries. This collective response of many receptors may function to emphasize predator information in the sensory system, and correlates with the amplitude range at which ultrasound elicits evasive behavior in tettigoniids. We compare our results with spectral processing in crickets, and discuss that both groups evolved different adaptations for the perceptual tasks of mate and predator detection.

Mid-frequency suppression in the green treefrog (Hyla cinerea): mechanisms and implications for the evolution of acoustic communication.

Advertisement calls of green treefrogs (Hyla cinerea) have two spectral peaks centered at about 1 kHz and 3 kHz. Addition of a component of intermediate frequency (1.8 kHz) to a synthetic call reduced its attractiveness to females relative to an alternative lacking this component. This mid-frequency suppression occurred over a 20-dB range of playback levels. Addition of other intermediate frequencies had weak effects on preferences at some playback levels, in some localities, and at lower-than-normal temperatures. These effects correlate well with the response properties of a population of low-frequency-tuned auditory neurons innervating the amphibian papilla. Males of a closely related species (H. gratiosa) produce calls with emphasized frequencies within the range of suppression in H. cinerea; however, suppression also occurred in localities well outside the area of geographical overlap with this species. Thus, previous speculation that mid-frequency suppression evolved to enhance species discrimination is probably incorrect. This phenomenon is more likely to reflect a general sensory bias in anurans and other vertebrates, tone-on-tone inhibition. Such negative biases, and other inhibitory mechanisms, almost certainly play an important role in the evolution of communication systems but have received far less attention than positive biases that enhance signal attractiveness.