Leader preference in Neoconocephalus ensiger katydids: a female preference for a nonheritable male trait.

Female preferences for males producing their calls just ahead of their neighbours, leader preferences, are common in acoustically communicating insects and anurans. While these preferences have been well studied, their evolutionary origins remain unclear. We tested whether females gain a fitness benefit by mating with leading males in Neoconocephalus ensiger katydids. We mated leading and following males with random females and measured the number and quality of F1 , the number of F2 and the heritability of the preferred male trait. We found that females mating with leaders and followers did not differ in the number of F1 or F2 offspring. Females mating with leading males had offspring that were in better condition than those mating with following males suggesting a benefit in the form of higher quality offspring. We found no evidence that the male trait, the production of leading calls, was heritable. This suggests that there is no genetic correlate for the production of leading calls and that the fitness benefit gained by females must be a direct benefit, potentially mediated by seminal proteins. The presence of benefits indicates that leader preference is adaptive in N. ensiger, which may explain the evolutionary origin of leader preference; further tests are required to determine whether fitness benefits can explain the phylogenetic distribution of leader preference in Neoconocephalus. The absence of heritability will prevent leader preference from becoming coupled with or exaggerating the male trait and prevent females from gaining a 'sexy-sons' benefit, weakening the overall selection for leader preference.

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.

Parallel changes in mate-attracting calls and female preferences in autotriploid tree frogs.

For polyploid species to persist, they must be reproductively isolated from their diploid parental species, which coexist at the same time and place at least initially. In a complex of biparentally reproducing tetraploid and diploid tree frogs in North America, selective phonotaxis--mediated by differences in the pulse-repetition (pulse rate) of their mate-attracting vocalizations--ensures assortative mating. We show that artificially produced autotriploid females of the diploid species (Hyla chrysoscelis) show a shift in pulse-rate preference in the direction of the pulse rate produced by males of the tetraploid species (Hyla versicolor). The estimated preference function is centred near the mean pulse rate of the calls of artificially produced male autotriploids. Such a parallel shift, which is caused by polyploidy per se and whose magnitude is expected to be greater in autotetraploids, may have facilitated sympatric speciation by promoting reproductive isolation of the initially formed polyploids from their diploid parental forms. This process also helps to explain why tetraploid lineages with different origins have similar advertisement calls and freely interbreed.

Molecular patterns of differentiation in canyon treefrogs (Hyla arenicolor): evidence for introgressive hybridization with the Arizona treefrog (H. wrightorum) and correlations with advertisement call differences.

Detection of genetic and behavioural diversity within morphologically similar species has led to the discovery of cryptic species complexes. We tested the hypothesis that US populations of the canyon treefrog (Hyla arenicolor) may consist of cryptic species by examining mate-attraction signals among three divergent clades defined by mtDNA. Using a multi-locus approach, we re-analysed phylogenetic relationships among the three clades and a closely related, but morphologically and behaviourally dissimilar species, the Arizona treefrog (H. wrightorum). We found evidence for introgression of H. wrightorum's mitochondrial genome into H. arenicolor. Additionally, the two-clade topology based on nuclear data is more congruent with patterns of call variation than the three-clade topology from the mitochondrial dataset. The magnitude of the call divergence is probably insufficient to promote isolation of the nuclear DNA-defined clades should they become sympatric, but further divergence in call properties significant in species identification could promote speciation in the future.

Divergence in the calling songs between sympatric and allopatric populations of the southern wood cricket Gryllus fultoni (Orthoptera: Gryllidae).

In the eastern United States the wood cricket Gryllus fultoni (Orthoptera: Gryllidae) occurs in sympatry with G. vernalis in an area between eastern Kansas and west of the Appalachian Mountains. Calling songs were recorded from 13 sympatric and allopatric localities. Both field and laboratory recordings showed that chirp rate (CR) and pulse rate (PR) overlapped extensively between allopatric populations of G. fultoni and sympatric populations of G. vernalis; by contrast, there was little or no overlap in these variables between sympatric populations of these two species. Divergence in PR and CR between the two species was thus greater in areas of sympatry than in areas of allopatry. Our field and laboratory studies of G. fultoni calling songs thus demonstrate the pattern expected of character displacement and support the genetic assumptions of this hypothesis. Other possible explanations for the sympatric divergence such as ecological character displacement and clinal variation are discussed.

AM representation in green treefrog auditory nerve fibers: neuroethological implications for pattern recognition and sound localization.

In addition to spectral call components, temporal patterns in the advertisement-call envelope of green treefrog males ( Hyla cinerea) provide important cues for female mate choice. Rapid amplitude modulation (AM) with rates of 250-300 Hz is typical for this species' advertisement calls. Here we report data on the encoding of these rapid call modulations by studying the responses of single auditory nerve fibers to two-tone stimuli with envelope periodicities close to those of the natural call. The free-field response properties of 86 nerve fibers were studied from 32 anesthetized males. The accuracy of stimulus envelope coding was quantified using both a Gaussian function fit to the interspike interval histograms derived from the first seven 20-ms stimulus segments, and the vector-strength metric applied to the phase-locked responses. Often, AM encoding in the initial stimulus segment was more faithful than that in its second half. This result may explain why conspecific females prefer calls in which the initial segment is unmasked rather than masked. Both the questions of pattern recognition and localization are discussed, and the data are related to behavioral observations of female choice and localization performance in this species.

Habituation as a mechanism of reduced aggression between neighboring territorial male bullfrogs (Rana catesbeiana).

Territorial animals often exhibit relatively lower levels of aggression toward familiar territorial neighbors than toward strangers. Habituation to a neighbor or its communication signals has been proposed to account for this reduced aggression between adjacent territorial neighbors. The authors asked whether discrimination between neighbors and strangers by territorial male bullfrogs (Rana catesbeiana) could result from habituation of the aggressive response to repeated presentations of the acoustic communication signals of a simulated new neighbor calling from an adjacent territory. In 3 field playback experiments, the authors found evidence for 5 response characteristics that operationally define habituation. Moreover, aggressive response decrements persisted between nights of chorusing and were specific to an individually distinct property of male advertisement calls. The authors suggest that reduced aggression between neighboring territorial male bullfrogs could result from long-term, stimulus-specific habituation to the advertisement calls of a new neighbor.

Polyploidy alters advertisement call structure in gray treefrogs.

Whole-genome duplication is believed to have played a significant role in the early evolution and diversification of vertebrate animals. The establishment of newly arisen polyploid lineages of sexually reproducing animals requires assortative mating between polyploids. Here, we show that genome duplication can directly alter a phenotypic trait mediating mate choice in the absence of genotypic change. Our results suggest that the direct effect of polyploidy on behaviour is a consequence of increased cell size.

Mating preference functions of individual female barking treefrogs, Hyla gratiosa, for two properties of male advertisement calls.

A mating preference function describes the relationship between variation in a trait in potential mates and the strength of the preference for that trait. Few studies have measured mating preference functions either at a population level or for individuals. We used two-choice playback experiments to determine the mating preference functions of individual female barking treefrogs for two call characteristics: call-repetition rate and fundamental frequency. We tested each female four times with each pair of stimuli and with three to six pairs of stimuli. Individual females exhibited directional preferences for higher call rates and stabilizing preferences for intermediate fundamental frequency. These individual preference functions were similar to population-level preferences documented in previous studies. Within a stimulus type (call rate or fundamental frequency), nearly all females exhibited the same general preference function. However, females varied in the minimum difference between stimuli necessary to elicit a unanimous choice for the higher call rate, and they differed in both the intermediate fundamental frequency they preferred most and the minimum difference in fundamental frequency that elicited a unanimous choice for one of the two alternatives. The variation we observed among females was not repeatable; repeatabilities were in general low and statistically nonsignificant. The observed variation in the preferences of females, even if unrepeatable, should weaken selection on male traits relative to selection in the absence of such variation.

A quantitative analysis of behavioral selectivity for pulse rise-time in the gray treefrog, Hyla versicolor.

The selectivity of female phonotactic responses to synthetic advertisement calls was tested in choice situations. Preferences based on differences in the linear rise-time of synthetic pulses depended on intensity and carrier frequency. When the carrier frequency was 1.1 kHz, simulating the low-frequency peak in the advertisement call, females preferred alternatives with slower rise-time pulses that differed by 5 ms at playback levels of 75 dB SPL and higher. A rise-time difference of 10 ms was discriminated at 65 dB SPL. When the carrier frequency was 2.2 kHz, simulating the high-frequency peak in the call, females discriminated a 5-ms difference in rise-time only at 85 dB SPL. Females showed no preference when the difference was 10 ms at lower playback levels. The difference in the thresholds (about 15-20 dB) for discriminating differences in rise-time at the two carrier frequencies was greater than the difference in behavioral thresholds for these two frequencies (about 10 dB). This result suggests that rise-time discrimination can be mediated solely by the neural channel mainly tuned to the low-frequency peak in the call. Females probably assess differences in rise-time by comparing the first few pulses of each call rather than by averaging over the entire call.

Isolation and characterization of simple sequence repeat loci in the gray tree frog, Hyla chrysoscelis.

A gray tree frog (Hyla chrysoscelis) genomic library was constructed and characterized with regard to the incidence and complexity of simple sequence repeat (SSR) loci. The partial genomic library, containing approximately 10,000 clones with an average-sized insert of 350 bp, was screened with six SSR repeat oligonucleotides (AC, AG, ACG, AGC, AAC, and AAG). Screening identified 31 unique positive clones containing 41 SSR loci. Sequences of tandemly arrayed dinucleotide repeats were more common (36 of 41) than trinucleotide repeats. Twenty-six loci were identified using the AC dinucleotide probe, while 7 loci were identified using the AG dinucleotide probe. An additional 3 AT dinucleotide loci were serendipitously identified. The AT repeats generally comprised the longest dinucleotide repeat loci. The SSR repeat loci reported here should provide potent markers for identity, parentage, and short-lineage determinations in large-scale experiments using gray tree frogs.

Significance of forebrain structures in acoustically guided behavior in anurans.

In order to test the hypothesis that the forebrain is involved in controlling acoustically guided behaviour, we carried out behavioural studies in combination with brain lesions, neuroanatomical and electrophysiological experiments in males and females of different species of frogs. Whereas the dorsomedial pallium plays no or only a minor role, the striatum, the septum, and the preoptic area potentially influence the behaviour because they send parallel descending projections to different premotor and motor networks in the brainstem. These parallel projections may be the basis for the variability seen in the behaviour.

Call duration as an indicator of genetic quality in male gray tree frogs.

The "good genes" hypothesis predicts that mating preferences enable females to select mates of superior genetic quality. The genetic consequences of the preference shown by female gray tree frogs for long-duration calls were evaluated by comparing the performance of maternal half-siblings sired by males with different call durations. Offspring of male gray tree frogs that produced long calls showed better performance during larval and juvenile stages than did offspring of males that produced short calls. These data suggest that call duration can function as a reliable indicator of heritable genetic quality.

Selective phonotaxis to advertisement calls in the grey treefrog Hyla versicolor: behavioral experiments and neurophysiological correlates.

1. The significance of particular acoustic properties of advertisement calls for selective phonotaxis by the gray treefrog, Hyla versicolor (= HV), was studied behaviorally and neurophysiologically. Most stimuli were played back at 85 dB SPL, a level typically measured at 1-2 m from a calling male. 2. Females preferred stimuli with conspecific pulse shapes at 20 degrees and 24 degrees C, but not at 16 degrees C. Tests with normal and time-reversed pulses indicated the preferences were not influenced by the minor differences in the long-term spectra of pulses of different shape. 3. Pulse shape and rate had synergistic or antagonistic effects on female preferences depending on whether the values of one or both of these properties in alternative stimuli were typical of those in HV or heterospecific (H. chrysoscelis = HC) calls. 4. More auditory neurons in the torus semicircularis were temporally selective to synthetic calls (90%) than to sinusoidally AM tones and noise (< 70%). 5. Band-pass neurons were tuned to AM rates of 15-60 Hz. Neurons were more likely to be tuned to HV AM rates (< 40 Hz) when stimuli had pulses with HV rather than HC shapes. 6. Sharp temporal tuning was uncommon and found only in neurons with band-pass or low-pass characteristics. 7. Many neurons differed significantly in response to HV and HC stimulus sets. Maximum spike rate was more often elicited by an HV stimulus (74%) than by an HC stimulus (24%). 8. Differences in spike rates elicited by HV and HC stimuli were attributable to combinations of differences in the rise times and shapes of the pulses.

Midbrain auditory sensitivity in the spring peeper (Pseudacris crucifer): correlations with behavioral studies.

1. We derived audiograms from recordings of multiunit activity in the torus semicircularis of 10 males and 6 females of the spring peeper from central Missouri, USA. We used free-field stimulation with tone bursts that had temporal properties similar to typical advertisement calls and that ranged in frequency from 500-6000 Hz. 2. Audiograms from different electrode positions in the same animal had the same general shape. There was no evidence of tonotopy. 3. Audiograms showed two regions of maximal sensitivity: a low-frequency region (500-700 Hz); and a high-frequency region (2000-4000 Hz). Absolute thresholds and frequencies of maximum sensitivity varied considerably from individual to individual. 4. Audiograms derived from all individuals of each sex indicated that in the high-frequency region, corresponding to the frequency range of advertisement calls, males were more broadly tuned than females. However, tuning in both sexes was relatively weak, and the data predict relatively little selectivity in behavioral responses over the entire range of variation in frequency of the advertisement call in local populations. 5. The results are discussed in terms of behavioral experiments with both males and females from the same populations in central Missouri. We show that merely summarizing the audiograms based on estimates of minimum thresholds of a population or species may mask significant individual differences in tuning. Moreover, most behavioral studies are conducted at playback levels considerably above threshold. For these reasons, behavioral selectivity is not always accurately predicted by inspection of "average" audiograms.

Directional hearing in the gray tree frog Hyla versicolor: eardrum vibrations and phonotaxis.

1. We used laser vibrometry to study the vibrational frequency response of the eardrum of female gray tree frogs for different positions of the sound source in three-dimensional space. Furthermore, we studied the accuracy of 3-D phonotaxis in the same species for sounds with different frequency contents. 2. The directionality of the eardrum was most pronounced in a narrow frequency range between 1.3 and 1.8 kHz. 3. The average 3-D, horizontal and vertical jump error angles for phonotactic approaches with a sound similar to the natural advertisement call (1.1 and 2.2 kHz frequency components) were 23 degrees, 19 degrees and 12 degrees, respectively. 4. 3-D jump error angle distributions for the 1.4 + 2.2 kHz, 1.0 kHz and 2.0 kHz sounds were not significantly different from that for the 1.1 + 2.2 kHz sound. 5. The average 3-D jump error angle for the 1.4 kHz sound was 36 degrees, and the distribution was significantly different from that for the 1.1 + 2.2 kHz sound. Hence, phonotactic accuracy was poorer in the frequency range of maximum eardrum directionality. 6. Head scanning was not observed and is apparently unnecessary for accurate sound localization in three-dimensional space. 7. Changes in overall sound pressure level experienced by the frog during phonotactic approaches are not an important cue for sound localization.

Female green treefrogs (Hyla cinerea) do not selectively respond to signals with a harmonic structure in noise.

1. Females of the green treefrog, Hyla cinerea, communicate in noisy environments, with spectrally complicated signals. A previous study (Megela Simmons 1988), using the reflex modification technique, found that the masked threshold of green treefrogs to two-tone signals differed by about 10 dB depending on whether or not the two components were harmonically-related. The present study used the same two-component stimuli to test the prediction that gravid females would better detect harmonic sounds in noise than inharmonic ones. 2. We offered gravid treefrogs simultaneous choices between alternative two-component synthetic sounds: (1) an inharmonic sound of 831 + 3100 Hz, and a harmonic sound of 828 + 2760 Hz. We varied the sound pressure level (SPL in decibels [dB]) to which we equalized these alternatives at the female's release point (75 and 80 dB SPL), and we tested females in quiet conditions and in the presence of broadband background noise (52 dB/Hz at the female's release point). 3. At a signal playback level of 75 dB SPL, one-third of the females responded in the presence of background noise. Subtracting the spectrum level yields a critical ratio estimate of 23 dB, a value that is very similar to estimates for single pure tones in noise reported in other studies of this species (Ehret and Gerhardt 1980; Moss and Megela Simmons 1986). Females did not, however, choose the harmonic sound over the inharmonic sound in this condition, at the higher signal-to-noise ratio, or in either of the unmasked situations.(ABSTRACT TRUNCATED AT 250 WORDS)

Recognition of spectral patterns in the green treefrog: neurobiology and evolution.

The natural communication behavior of frogs has provided a framework for studying both the mechanisms and evolution of sound pattern recognition. In the green treefrog behavioral studies using synthetic stimuli have identified the pertinent acoustic properties of its complex but stereotyped vocal signals. There are two optimal frequency bands: the most effective signals have about the same amount of energy in the two bands. Neurophysiological studies revealed biases for these two frequency bands at the level of the peripheral auditory system. Furthermore, the neural response properties in an auditory thalamic area are especially well correlated with selective phonotaxis based on the appropriate combination of low- and high-frequency sound energy. Comparing the optimal frequency bands in terms of female preferences with the range of frequencies produced by males in natural populations suggests that intraspecific mate choice based on call frequency alone is unlikely.