Pre-existing sensory biases in the spectral domain in frogs: empirical results and methodological considerations

In many species of anurans, advertisement calls excite only one of the two inner-ear organs. One prediction of the pre-existing bias hypothesis is that signal innovations that additionally excite the “untapped” organ will be more behaviorally effective than normal calls. However, recent studies have shown that females of three species with single-peaked calls that stimulate only the basilar papilla (BP) preferred single-peaked synthetic calls with a frequency typical of conspecific calls to two-peaked calls that also stimulated the amphibian papilla (AP). We report that in spring peepers (Pseudacris crucifer) that also produce single-peaked calls, females did not show a preference in choices between single-peaked and two-peaked synthetic calls. Thus, the addition of energy exciting the AP had a neutral effect on signal attractiveness. Together, these results are unsupportive of the pre-existing bias hypothesis. An alternative hypothesis is that positive fitness consequences of responding to sounds providing extraordinary spectral stimulation are required for a novel call to become established as a mate-attracting signal. Testing these ideas requires a taxonomically broader examination of responses to sounds with novel spectral complexity, and attention to some methodological details will improve the comparability of such studies.     

Temporal call changes and prior experience affect graded signalling in the cricket frog

We investigated how male cricket frogs Acris crepitans, alter their advertisement calls in response to broadcasts of synthetic calls that were either 'attractive' or 'aggressive'. The stimulus calls differed in temporal but not spectral characteristics. Male cricket frogs produced a more aggressive call when presented with the aggressive stimulus, indicating that they perceived the temporal differences between the two call categories. The direction and degree of temporal and spectral changes depended on the relative dominant frequency of the resident and opponent. If the resident's dominant frequency was initially higher than the stimulus frequency, the pattern of change in dominant frequency mirrored that seen for the temporal call characters. In contrast, if the resident's initial dominant frequency was below that of the stimulus, then the temporal and spectral changes were in opposite directions. Furthermore, stimulus order influenced whether males responded differently to playbacks of aggressive and attractive calls; males that received the aggressive call first produced more aggressive calls during the aggressive stimulus, while males that received the attractive call first produced similar calls in response to the two stimuli. This suggests that experience with different types of signals influences the subsequent calling behaviour of male cricket frogs. Copyright 1999 The Association for the Study of Animal Behaviour.     

Female reproductive state influences the auditory midbrain response

Female behavioral responses to sensory stimuli can be highly variable across the reproductive cycle. Female green treefrogs (Hyla cinerea) use the male vocal signal to locate and choose a mate. Gravid females approach a vocalizing male to mate but do not approach if they have recently mated. Such differences in behavioral response may be due in part to shifts in the neural representation of auditory information in the brain. In this study, we investigated the influence of female reproductive state on neural responses in the auditory midbrain to both communication signals (advertisement calls) and non-communication sounds (band limited noise bursts). Recently mated females exhibited significantly reduced response strengths compared to females not recently mated. Reduced response strengths in post-mated females were in response to both noise bursts and male advertisement calls but were limited to the lower frequency range corresponding to the amphibian papilla of the peripheral auditory system. Our results therefore show that the ability of social signals to stimulate the auditory system differs in females depending on their reproductive state, and that the differential effect on low versus high spectral sensitivities may influence the way the two spectral peaks of male advertisement calls are represented.     

Signal Redundancy and Receiver Permissiveness in Acoustic Mate Recognition by the Tungara Frog, Physalaemus pustulosus

The tungara frog has an advertisement call with two structurallyand functionally distinct components: the whine is both necessaryand sufficient for species recognition, and addition of chucksfurther enhances call attractiveness. Only the fundamental frequencyof the whine contributes to phonotaxis; the upper harmonicsplay no role. Furthermore, only a small portion of the whine,within the first 0–100 msec, is necessary to elicit phonotaxis;there is some redundancy since either the 0–50 msec orthe 50–100 msec portion elicits a response. Most of theremainder of the whine's fundamental further enhances call attractiveness.Only the amphibian papilla in the peripheral auditory systemis involved in decoding the whine. In stark contrast, severalchuck variants, which stimulate either the amphibian papillaor the basilar papilla, and white noise are as effective inenhancing call attractiveness as is the normal chuck, showingthat either peripheral end organ can be involved in the perceptionof a more attractive call. Thus there is greater receiver specificityfor decoding stimuli in interspecific mate choice, and greaterreceiver permissiveness for decoding stimuli in intraspecificmate choice; it appears that intraspecific mate choice can takeadvantage of a greater array of neural pathways for call decodingthan can interspecific mate choice.     

Sexual signaling and immune function in the black field cricket Teleogryllus commodus

The immunocompetence handicap hypothesis predicts that male sexual trait expression should be positively correlated with immunocompetence. Here we investigate if immune function in the cricket, Teleogryllus commodus, is related to specific individual components of male sexual signals, as well as to certain multivariate combinations of these components that females most strongly prefer. Male T. commodus produce both advertisement and courtship calls prior to mating. We measured fine-scale structural parameters of both call types and also recorded nightly advertisement calling effort. We then measured two standard indices of immune function: lysozyme-like activity of the haemolymph and haemocyte counts. We found a weak, positive relationship between advertisement calling effort and lysozyme-like activity. There was, however, little evidence that individual structural call components or the net multivariate attractiveness of either call type signalled immune function. The relationships between immunity and sexual signaling did not differ between inbred and outbred males. Our data suggest that it is unlikely that females assess overall male immune function using male calls.     

Double stimulation of the inner ear organs of an anuran species (Alytes cisternasii) with simple tonal advertisement calls

Midwife toads present one of the simplest calls in anurans, with the whole energy concentrated in a single band without frequency modulation. The tuning curves of the Iberian midwife toads Alytes cisternasii show the typical bimodal pattern in anurans, with two best excitatory frequencies at 0.412 kHz (corresponding to the amphibian papilla) and at 1.358 kHz (corresponding to the basilar papilla and matching the male call frequency). In this study, the hypothesis that complex calls arose in anurans because they were inherently more attractive to females, since they provided greater acoustic stimulation, was tested. However, our results indicate that splitting the call energy to stimulate both inner ear organs simultaneously, the male call is not more attractive to female midwife toads, but sometimes renders it unattractive. The biological role of the amphibian papilla is discussed in ecological and evolutionary terms.     

Mechanics of the inner ear of the bullfrog (Rana catesbeiana): the contact membranes and the periotic canal

The frog inner ear consists of a complex of fluid-filled membranous sacs and canals containing eight distinct clusters of sensory hair cells. In this study we attempt to delineate the potential pathways for acoustic energy flow toward two of these clusters located within the amphibian papilla and the basilar papilla. Detailed morphological measurements of the periotic canal based on internal casts of the inner ear in the bullfrog (Rana catesbeiana) revealed that it is divided into a wide, tapered section and a narrower section comprised of two branches – one short and blind projecting into the endolymphatic space and another longer, terminating in the round window. Additionally, we used laser Doppler velocimetry to record the velocity responses of the contact membranes of the amphibian papilla and basilar papilla. We found that the acoustic energy flow through these two structures is frequency dependent such that the amphibian papilla contact membrane displays a peak velocity amplitude at frequencies less than 500 Hz, whereas the basilar papilla contact membrane velocity response exhibits a maximum above 1100 Hz. Our data advocate a mechanical substrate underlying the frequency segregation in the auditory nerve fibers innervating the amphibian papilla and the basilar papilla. Accepted: 9 March 2000     

Arginine Vasotocin Injection Increases Probability of Calling in Cricket Frogs, but Causes Call Changes Characteristic of Less Aggressive Males

Male cricket frogs,Acris crepitanscommunicate to males and females using advertisement calls, which are arranged into call groups. Calls at the middle and end, but not beginning of the call group, are modified in response to male–male aggressive interactions. We found in this field study of male cricket frogs in natural breeding choruses that the peptide hormone arginine vasotocin (AVT) not only increased the probability that males called after injections, but also caused modifications in middle and end calls to produce calls characteristic of less aggressive males. Moreover, AVT-injected males showed significantly greater increases in call dominant frequency than saline-injected males, again, a characteristic of less aggressive males. Cricket frog calls are used to both repel males and attract females, thus call changes may relate to male–male and/or male–female interactions. Saline-injected males also demonstrated significant changes in several call traits, including changes that occurred in the beginning and middle calls of the call groups, but not the end calls. AVT appeared to block some call changes produced through handling. These data suggest that AVT can influence acoustic communication in frogs in several ways, including effects on call characteristics and dominant frequency, as well as potentially blocking some handling effects.     

THE SENSORY BASIS OF SEXUAL SELECTION FOR COMPLEX CALLS IN THE TÚNGARA FROG,PHYSALAEMUS PUSTULOSUS (SEXUAL SELECTION FOR SENSORY EXPLOITATION)

Male túngara frogs (Physalaemus pustulosus) vocalize to attract females, and enhance the attractiveness of their simple, whine-only call by adding chucks to produce complex calls. Complex calls contain more total energy and are of longer duration. By virtue of the greater frequency range of the chuck, complex calls also simultaneously stimulate both the amphibian papilla and the basilar papilla of the frog's inner ear. Female phonotaxis experiments using synthetic stimuli demonstrate that an increase in the call's acoustic energy is not sufficient to account for the enhanced attractiveness of the complex call. However, the stimulation of either or both of the female's sound-sensitive inner-ear organs is sufficient to elicit her preference. We suggest that the female's sensory system generates selection that equally favors at least three evolutionary alternatives for enhancing call attractiveness and that historical constraints imposed by the male's morphology determined which of the alternatives was more likely to evolve. These data are consistent with our hypothesis of sensory exploitation, which states that selection favors those traits that elicit greater stimulation from the female's sensory system and which emphasizes the nonadaptive nature of female preference.     

The separate and combined effects of harmonic structure, phase, and FM on female preferences in the barking treefrog (Hyla gratiosa)

In natural advertisement calls of the barking treefrog, Hyla gratiosa, a small amount of incoherent frequency modulation (FM) is present. Incoherency in the FM of a call creates inharmonicity and phase changes between its frequency components. In this study, the combined and separate effects of the harmonic structure, phase spectrum, and FM of an advertisement call on female choice were tested. The harmonic structure of a call can have a direct effect on female preference; females showed a significant preference for static-inharmonic calls over static-harmonic calls. Neither differences in phase or FM alone conferred a preference in two choice tests. However, when FM is present in both calls it does influence female preference for harmonic structure -namely harmonic calls become preferable to inharmonic calls. This reversal of female preference for inharmonicity in a call by the presence of FM suggests that call parameters may interact, and thereby effect mate choice.Abbreviations AP amphibian papilla - BP basilar papilla - FM frequency modulation - PM phase modulation - HS harmonic structure - GB Gaussian Band     

A laryngeal fibrous mass impacts the acoustics and attractiveness of a multicomponent call in túngara frogs (Physalaemus pustulosus)

The evolution of elaborate signals can emerge from changes in anatomical signaling structures. In the male túngara frog, a simple advertisement call (‘whine’) can be ornamented facultatively with a suffix (‘chuck’) to produce a more attractive complex call, or ‘whine-chuck’. A fibrous mass (FM1) supported by the vocal cords plays a role in chuck production. Here, we examine the effects of FM1 ablation on a large set of spectral and temporal features of both the whine and chuck and we test the hypothesis that FM1 ablation reduces call attractiveness to females. Both call components were impacted by FM1 ablation, but especially the suffix. The proportion of energy in the suffix’s odd compared to even harmonics diminished markedly as did the relative amplitude, effectively eliminating the chuck percept. FM1 ablation also reduced the whine’s frequency and its rate of frequency modulation. Moreover, post-surgery chucks no longer enhanced the attractiveness of the simple call and whines also appeared to diminish in attractiveness. Together, our results demonstrate that FM1 plays a role in the production of both call components in a way that stimulates the female auditory system and that the growth of FM1 had potentially positive sexual selection implications for the pre-existing simple call.     

Diversity of form in the amphibian papilla of Puerto Rican frogs

Summary In modern frogs, the amphibian papilla exhibits a caudal extension whose shape, relative length, and proportion of hair cells vary markedly from species to species. Tuning in the caudal extension is organized tonotopically and evidently involves the tectorium. In terms of the proportion of amphibian-papillar hair cells in the caudal extension, we report more diversity among 8 species of a single genus (Eleutherodactylus) on a single island (Puerto Rico) than has been found so far among all of the (more than 50) other modern anurans examined for this feature from around the world. These 8 Puerto Rican species have overlapping habitat and conspicuous diversity in the male advertisement call. For 7 of the 8 species, we report that the call has transient spectral components in the frequency range of the amphibian papilla, and that the proportion of caudal extension hair cells and the frequency distribution of those components are correlated. Thus one might conclude that the selective pressures that led to diversity of calls among the 8 species also led to diversity in form of the amphibian papilla.Abbreviations AP amphibian papilla - BEF best excitatory frequency - PR Puerto Rican - SEM scanning electron microscope - SVL snout-vent length - TC tectorial corner - TM tectorial membrane     

Correlation between auditory evoked responses in the thalamus and species-specific call characteristics

This evoked potential study of the bullfrog's auditory thalamic area (an auditory responsive region in the posterior dorsal thalamus) shows that complex processing, distinct from that reported in lower auditory regions, occurs in this center. An acoustic stimulus consisting of two tones, one which stimulates either the low-frequency or the mid-frequency sensitive population of auditory nerve fibers from the amphibian papilla and the other the high-frequency sensitive population of fibers from the basilar papilla, evoked a maximal response. The amplitude of the response to the simultaneous stimulation of the two auditory organs was, in some locations, much larger than the linear sum of the responses to the individual tones presented separately. Bimodal spectral stimuli that had relatively long rise-times (greater than or equal to 100 ms) evoked much larger responses than similar sounds with short rise-times. The optimal rise-times were close to those occurring in the bullfrog's mating call. The response was dependent on the waveform periodicity and harmonic content, with a fundamental frequency of 200 Hz producing a larger response than those with fundamentals of 50, 100 or 300 Hz. Six of the natural calls in the bullfrog's vocal repertoire were tested and the mating call and warning call were found to evoke the best responses. Each of these calls stimulate the two auditory organs simultaneously. The evoked response had a long refractory period which could not be altered by lesioning the efferent telencephalic pathways. The type of spectral and temporal information extracted by the auditory thalamic area suggests that this center is involved in processing complex sounds and likely plays an important role in the bullfrog's detection of some of its vocal signals.     

The rôle of call frequency and the auditory papillae in phonotactic behavior in male Dart-poison frogs <Emphasis Type="Italic">Epipedobates femoralis</Emphasis> (Dendrobatidae)

Territorial males of the pan-Amazonian Dart-poison frog, Epipedobates femoralis, are known to present stereotypic phonotactic responses to the playback of conspecific and synthetic calls. Fixed site attachment and a long calling period within an environment of little temperature change render this terrestrial and diurnal pan-Amazonian frog a rewarding species for field bioacoustics. In experiments at the field station Aratai, French Guiana, we tested whether the prominent frequency modulation of the advertisement-call notes is critical for eliciting phonotactic responses. Substitution of the natural upward sweep by either a pure tone within the species frequency range or a reverse sweep did not alter the males' phonotactic behavior. Playbacks with artificial advertisement calls embedded in high levels of either low-pass or high-pass masking noise designed to saturate nerve fibers from either the amphibian papilla or basilar papilla showed that male phonotactic behavior in this species is subserved by activation of the basilar papilla of the inner ear.     

Hearing conspecific vocal signals alters peripheral auditory sensitivity

We investigated whether hearing advertisement calls over several nights, as happens in natural frog choruses, modified the responses of the peripheral auditory system in the green treefrog, Hyla cinerea. Using auditory evoked potentials (AEP), we found that exposure to 10 nights of a simulated male chorus lowered auditory thresholds in males and females, while exposure to random tones had no effect in males, but did result in lower thresholds in females. The threshold change was larger at the lower frequencies stimulating the amphibian papilla than at higher frequencies stimulating the basilar papilla. Suprathreshold responses to tonal stimuli were assessed for two peaks in the AEP recordings. For the peak P1 (assessed for 0.8–1.25 kHz), peak amplitude increased following chorus exposure. For peak P2 (assessed for 2–4 kHz), peak amplitude decreased at frequencies between 2.5 and 4.0 kHz, but remained unaltered at 2.0 kHz. Our results show for the first time, to our knowledge, that hearing dynamic social stimuli, like frog choruses, can alter the responses of the auditory periphery in a way that could enhance the detection of and response to conspecific acoustic communication signals.     

Baffling: a condition-dependent alternative mate attraction strategy using self-made tools in tree crickets

Intense sexual selection in the form of mate choice can facilitate the evolution of different alternative reproductive strategies, which can be condition-dependent. Tree cricket males produce long-distance acoustic signals which are used by conspecific females for mate localization and mate choice. Our study shows that baffling, an acoustic call amplification strategy employed by male tree crickets using self-made tools, is a classic example of a condition-dependent alternative strategy. We show that though most males can baffle, less preferred males, such as smaller and lower-amplitude callers, predominantly use this alternative strategy. Baffling allows these males to increase their call amplitude and advertisement range, which attracts a higher number of females. Baffling also gives these males a mating benefit because females mate for longer durations with them. Our results suggest that the advantage of baffling in terms of gain in the number of sperm cells transferred while mating is primarily limited to less preferred males, thus maintaining the polymorphism of calling strategies in the population. We summarize that baffling is a condition-dependent strategy used by less preferred tree cricket males to obtain mating benefits.     

Acoustic,auditory, and morphological divergence in three species of neotropical frog

Advertisement calls, auditory tuning, and larynx and ear morphology were examined in 3 neotropical frogs, Hyla microcephala, H. phlebodes and H. ebraccata, H. microcephala has the highest call dominant frequency (6.068 kHz) and basilar papilla tuning (5.36 kHz). H. phlebodes and H. ebraccata calls have lower dominant frequencies (3.832 and 3.197 kHz respectively) and basilar papilla tuning (2.79 and 2.56 kHz). The primary call notes of H. ebraccata are longer (181.6 ms) than those of H. microcephala (95.5 ms) or H. phlebodes (87.3 ms). Morphometric analysis suggests that temporal call features differ as laryngeal musculature changes, in the process changing the overall size of the larynx. The spectral aspects of the call differ as head size, and hence the size of its resonating and radiating structures, changes, modifying the dominant frequency of calls by accentuating their higher harmonics when head size decreases. Decreasing head size decreases the size of the middle and inner ear chambers, changing the mechanical tuning of the ear in the same direction as the change in dominant frequency. These changes result in divergent spectral-temporal characteristics of both the sending and receiving portions of the acoustic communication system underlying social behavior in these frogs.Abbreviations AP amphibian papilla - BEF best excitatory frequency - BP basilar papilla - dB SPL decibels sound pressure level re:20 N/m²     

Acoustically induced call modification in the white-lipped frog,Leptodactylus albilabris

The vocalization behaviour of Leptodactylus albilabris was investigated using field playback experiments. To assess the response of males to pre-recorded natural ‘chirp’ (advertisement call) and natural ‘chuckle” (aggressive call) stimuli of gradually increasing broadcast intensity, three parameters (intensity, dominant frequency and repetition rate) of the chirp call were analysed. Of the males tested, 69% showed a significant increase in chirp intensity with increased levels of both stimulus types. In response to playback of the chirp stimulus, males actively modified the dominant frequency of their chirp calls over a mean range of 91·42 Hz, and in one case as much as 400 Hz. Moreover, 12 of 17 males shifted the frequency of their call towards the dominant frequency of the chirp stimulus (2175 Hz) by either increasing or decreasing the dominant frequency of their chirp calls. In response to the natural chuckle stimulus, 83% of the males showed either a decrease or no significant change in the dominant frequency of their chirps. All eight males for which both the chirp frequency and intensity were analysed and that showed an increase in chirp intensity also showed a concomitant increase in chirp dominant frequency. These results are the first to document quantitatively the plasticity of advertisement call intensity and dominant frequency in an anuran. The possible effects of advertisement call modification on male mating success in L. albilabris is discussed.     

Mid-frequency suppression in the green treefrog (<Emphasis Type="Italic">Hyla cinerea</Emphasis>): 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.     

Tympanic and extratympanic sound transmission in the leopard frog

Summary The inner ear of the leopard frog,Rana pipiens, receives sound via two separate pathways: the tympanic-columellar pathway and an extratympanic route. The relative efficiency of the two pathways was investigated. Laser interferometry measurements of tympanic vibration induced by free-field acoustic stimulation reveal a broadly tuned response with maximal vibration at 800 and 1500 Hz. Vibrational amplitude falls off rapidly above and below these frequencies so that above 2 kHz and below 300 Hz tympanic vibration is severely reduced. Electrophysiological measurements of the thresholds of single eighth cranial nerve fibers from both the amphibian and basilar papillae in response to pure tones were made in such a way that the relative efficiency of tympanic and extratympanic transmission could be assessed for each fiber. Thresholds for the two routes are very similar up to 1.0 kHz, above which tympanic transmission eventually becomes more efficient by 15–20 dB. By varying the relative phase of the two modes of stimulation, a reduction of the eighth nerve response can be achieved. When considered together, the measurements of tympanic vibration and the measurements of tympanic and extratympanic transmission thresholds suggest that under normal conditions in this species (1) below 300 Hz extratympanic sound transmission is the main source of inner ear stimulation; (2) for most of the basilar papilla frequency range (i.e., above 1.2 kHz) tympanic transmission is more important; and (3) both routes contribute to the stimulation of amphibian papilla fibers tuned between those points. Thus acoustic excitation of the an uran's inner ear depends on a complex interac tion between tympanic and extratympanic sound transmission.Abbreviations dB SPL decibels sound pressure level re: 20 N/ m² - AP amphibian papilla - BP basilar papilla - BEF best excitatory frequency