Social context-induced song variation affects female behavior and gene expression

Social cues modulate the performance of communicative behaviors in a range of species, including humans, and such changes can make the communication signal more salient. In songbirds, males use song to attract females, and song organization can differ depending on the audience to which a male sings. For example, male zebra finches (Taeniopygia guttata) change their songs in subtle ways when singing to a female (directed song) compared with when they sing in isolation (undirected song), and some of these changes depend on altered neural activity from a specialized forebrain-basal ganglia circuit, the anterior forebrain pathway (AFP). In particular, variable activity in the AFP during undirected song is thought to actively enable syllable variability, whereas the lower and less-variable AFP firing during directed singing is associated with more stereotyped song. Consequently, directed song has been suggested to reflect a “performance” state, and undirected song a form of vocal motor “exploration.” However, this hypothesis predicts that directed–undirected song differences, despite their subtlety, should matter to female zebra finches, which is a question that has not been investigated. We tested female preferences for this natural variation in song in a behavioral approach assay, and we found that both mated and socially naive females could discriminate between directed and undirected song—and strongly preferred directed song. These preferences, which appeared to reflect attention especially to aspects of song variability controlled by the AFP, were enhanced by experience, as they were strongest for mated females responding to their mate's directed songs. We then measured neural activity using expression of the immediate early gene product ZENK, and found that social context and song familiarity differentially modulated the number of ZENK-expressing cells in telencephalic auditory areas. Specifically, the number of ZENK-expressing cells in the caudomedial mesopallium (CMM) was most affected by whether a song was directed or undirected, whereas the caudomedial nidopallium (NCM) was most affected by whether a song was familiar or unfamiliar. Together these data demonstrate that females detect and prefer the features of directed song and suggest that high-level auditory areas including the CMM are involved in this social perception.     

Social Context–Induced Song Variation Affects Female Behavior and Gene Expression

Social cues modulate the performance of communicative behaviors in a range of species, including humans, and such changes can make the communication signal more salient. In songbirds, males use song to attract females, and song organization can differ depending on the audience to which a male sings. For example, male zebra finches (Taeniopygia guttata) change their songs in subtle ways when singing to a female (directed song) compared with when they sing in isolation (undirected song), and some of these changes depend on altered neural activity from a specialized forebrain-basal ganglia circuit, the anterior forebrain pathway (AFP). In particular, variable activity in the AFP during undirected song is thought to actively enable syllable variability, whereas the lower and less-variable AFP firing during directed singing is associated with more stereotyped song. Consequently, directed song has been suggested to reflect a “performance” state, and undirected song a form of vocal motor “exploration.” However, this hypothesis predicts that directed–undirected song differences, despite their subtlety, should matter to female zebra finches, which is a question that has not been investigated. We tested female preferences for this natural variation in song in a behavioral approach assay, and we found that both mated and socially naive females could discriminate between directed and undirected song—and strongly preferred directed song. These preferences, which appeared to reflect attention especially to aspects of song variability controlled by the AFP, were enhanced by experience, as they were strongest for mated females responding to their mate's directed songs. We then measured neural activity using expression of the immediate early gene product ZENK, and found that social context and song familiarity differentially modulated the number of ZENK-expressing cells in telencephalic auditory areas. Specifically, the number of ZENK-expressing cells in the caudomedial mesopallium (CMM) was most affected by whether a song was directed or undirected, whereas the caudomedial nidopallium (NCM) was most affected by whether a song was familiar or unfamiliar. Together these data demonstrate that females detect and prefer the features of directed song and suggest that high-level auditory areas including the CMM are involved in this social perception.     

Male singing behaviour and female presence in the territory in whitethroats Sylvia communis

Unmated male songbirds usually change their vocal behaviour when females enter their territories. Either the males court the females by changing the rate or pattern with which their normal long-ranging full songs are emitted, or they shift to special displays and long- or short-ranging vocalisations. In this study we quantified how female presence and behaviour affect the singing behaviour of male whitethroats. In the presence of a female the male frequently performed song flights, maybe to locate the female before it was courted, with sequences of diving-song displays. The courtship was interrupted by periods of perch songs. Female dscharp calls and short movements made the males initiate or resume courtship, whereas short horizontal jumps made the males intensify their courtship. Overall, the males changed their dual-function song activity in females' presence by emitting fewer perch songs and more flight songs. The quiet diving songs were only emitted during direct courtship of a female. The song types emitted immediately before, during, and after courtship are all highly variable, thus allowing for a quick assessment of the male's song repertoire. The courtship was also interrupted by periods of male woid calling, a call that is known to have a deterring effect on rival males. Bouts of woid calls were usually followed by song flights, again probably to locate the female that might have moved out of sight, or maybe to locate potential rival intruders. The latter was supported by an increased intrusion rate during female presence. Communicated by P. McGregor     

Context-dependent effects of castration and testosterone treatment on song in male European starlings

Most seasonally breeding songbirds display dramatic seasonal fluctuations in plasma testosterone (T) levels and mate attraction behaviors, including song. However, males of some songbird species, such as the European starling (Sturnus vulgaris), continue to sing at high levels after the breeding season, when T levels are basal. In male starlings song during the breeding season functions mainly to attract mates, whereas song during the nonbreeding season appears unrelated to reproduction. This suggests that song produced in a context unrelated to female courtship, unlike song directed toward females, is not regulated by plasma T. In captive males housed in large outdoor aviaries we explored the relationship between plasma T and song produced during the breeding season within and outside a courtship context. This was achieved by determining the effects of castration and subsequent T treatment on song and mate attraction behaviors in both the presence and the absence of a female. Compared to intact males, castrated males did not show reduced song activity in the absence of a female for at least 6 months after the operation, strongly suggesting that the expression of noncourtship song is not regulated by plasma T. Likewise, we found that experimentally elevating T levels in castrated males did not affect noncourtship song rates. However, control castrated males receiving empty implants tended to show reduced noncourtship song rates after implantation. This may have been due to a suppressive effect caused by the presence of the T-implanted castrated males in the same aviary. In contrast, courtship singing was clearly controlled by plasma T: it was abolished by castration and restored by subsequent T replacement when males were housed both individually and in a group situation. High plasma levels of T also appeared necessary for the activation of three other behavioral traits critical for mate attraction, namely, nesthole occupancy, spending time (singing) in a nesthole, and carrying green nesting material into a nesthole.     

Watch Your Tone: Social Conditions Modulate Singing Strategies

Bird song is typically depicted as a male singing a long‐distance signal to potentially unknown receivers to (1) deter males and (2) attract females. Nevertheless, many songbirds sing from close distances to a known receiver; males of these species may be under more intense selective pressure to modify their songs depending on the sex of the receiver in order to convey different motivational states (aggression versus courtship) to the different sexes. In a laboratory setting, we examined how receiver sex affected within‐song variation of the close‐range singing behavior in the brown‐headed cowbird (Molothrus ater). Although we know that cowbird song is influenced by flock composition, it is still unclear as to how the cowbird modifies his song based on social context. Using a cross‐correlation analysis of each male's different song types, we found that pairs of songs were significantly more dissimilar if they were directed to females compared with songs directed to males. We subsequently tested whether there were any consistent spectral or temporal patterns in the songs males gave to females versus to males. Our results lend support for the Motivational Structural Rules Hypothesis as songs directed toward males had higher entropy (i.e., harshness) than the same song type directed toward females. Our results suggest that cowbirds may have evolved the ability to alter multiple dimensions of their singing behavior based on receiver sex.     

The song structure and seasonal patterns of vocal behavior of male and female bellbirds (Anthornis melanura)

The bellbird (Anthornis melanura) is a honeyeater endemic to New Zealand, which uses song to defend breeding territories and/or food resources year round. Both sexes sing and the song structure and singing behavior have not yet been quantified. The number of song types, spectral structure, repertoire size, and singing behavior of male and female bellbirds was investigated for a large island population. Song types differed between the sexes with males singing a number of structurally distinct song types and females producing song types that overlapped in structure. Singing behavior also differed between the sexes; males often sung long series of songs while females sung each song at relatively long and variable intervals. Singing by both sexes occurred year round but the frequency of male and female singing bouts showed contrasting seasonal patterns. The frequency of female singing bouts increased as the breeding season progressed, whereas male singing bouts decreased. In contrast to almost all studied passerines, female bellbirds exhibited significant singing behavior and sung songs of complex structure and variety that parallel male song. These results provide a quantitative foundation for further research of song in bellbirds and in particular the function of female vocal behavior.     

丹顶鹤性活动的声行为研究

丹顶鹤繁殖期的性活动可分为雄鹤求偶、雌鹤对雄性求偶的应答、两性交配和交配完结4个阶段,其相应的鸣声模式分别为雄性的求偶鸣声、雌性对雄性求偶的应答声和两性的对鸣声、两性对唱的交配声和两性的高声合唱。4个阶段鸣声都是以基本音的主频率(PF)为主音的单音调声,前3个阶段都带数个近似fn=nf0(f0=FP)关系的低幅值谐频成分。第4个阶段带数个近似fn=nf0(f0=FP)关系的高幅值谐频成分;品质因数(Q3dB)多半为4～6,声脉冲重复频率(RFP)一般为150～180Hz,而第2阶段声的RFP一般为180～260Hz。雄性鸣声的每个单次叫声中含有的音节数较少,一般不超过4个;而雌性鸣声比较复杂。每个单次叫声中含有的音节数较多,一般都在7～8个以上;但雌雄鸣声的每个音节都是由3个声脉冲组成。雄鹤鸣唱声频率变化范围较小,而雌鹤鸣唱声频率变化形式是由低到高达到高峰后又开始下降。4个阶段的鸣声都具有较好共鸣。只有第2阶段发声运动较快。而且发现雄鹤鸣唱单次鸣叫声的音节数“增多”。各阶段鸣声特性均存在差异,不同配偶间均存在显著差异,研究结果表明丹顶鹤雌雄都具有不同的鸣声,且其性活动过程中不同的鸣声行为具有较高的个体识别信号潜能。另外,求偶鸣叫声和求偶应答与对鸣声在性活动鸣声中起着决定性的作用。     

Broad Selectivity for Courtship Song in the Cricket Gryllus bimaculatus

Various characteristics of a long‐distance acoustic signal have been shown to vary to different degrees. It has been suggested that female preferences based on stable song parameters are stabilising or weakly directional, and preferences based on variable parameters are strongly directional. We tested this hypothesis based on a short‐distance signal (courtship song) produced by the field cricket, Gryllus bimaculatus. We studied the degree of variability of different courtship song parameters and the behavioural importance of several parameters using synthesised song models in playback experiments. We found that most of the courtship song elements of G. bimaculatus were quite variable (coefficient of variation, CV, in the range of 20–53%). The most variable parameter of the courtship song was the relative amplitude of two elements: high‐amplitude ticks and low‐amplitude pulses. Because songs containing only ticks (of rare occurrence) appeared to be more effective than songs with both ticks and pulses (of frequent occurrence), we consider female preferences to be directional. Alteration of less variable traits, such as the carrier frequency and duration of ticks (CV = 20–25%), had a different effect on female responsiveness. The synthesised songs with different carrier frequencies of ticks were as attractive to females as the positive control (courtship of muted males accompanied by playback of the recorded song). Altering the duration of ticks had a crucial effect on the female response rate, decreasing female responsiveness to the level observed in the negative control (courtship of muted males). Thus, we did not find a strong relationship between the variability of individual song parameters and their potential importance in song recognition and the evaluation of male quality. The partial inconsistency of our results with the data of other authors may be due to different patterns of past and current selection on long‐distance and short‐distance acoustic signals.     

Variance in female responses to the fine structure of male song in the field cricket, Gryllus integer

Although female mating preferences are a focus of current controversy,little detailed information exists on female preferences withinnatural populations. In the field cricket Gryllus integer, malecalls attract sexually receptive females, and females preferentiallymove toward male calls with longer calling bouts (periods ofcalling containing no pause greater than 0.10 s in real time).This study investigated female preferences for other variablesof the male song, including syllable period, chirp pause, andnumber of syllables per chirp. Male song was measured in thefield to determine mean values for each variable in nature.Female preferences were determined using a locomotor-compensatordevice, on which females ran in response to sequential playbacksof synthesized male song. Mean female preferences correspondedroughly to mean male song variables. Nonetheless, females variedgreatly in their responses to synthesized calls differing insyllable period, syllable number, and chirp pause. Moreover,individual females who were more selective for any one variablealso tended to be more selective for others. These results showthat females may differ from one another in their mating preferencesand degrees of selectivity, even within a single population.     

The importance of calling song and courtship song in female mate choice in the variable field cricket

Male field crickets produce calling songs, courtship songs, tactile signals and chemical signals. Although calling songs are known to play an important role in female mate choice, the importance of the other signals in mate choice is poorly understood. In the variable field cricket, Gryllus lineaticeps, females select mates, in part, based on variation in male calling song. Females prefer higher chirp rates, a trait which is partially dependent on male nutrient intake, and females prefer longer chirp durations, a trait which appears to be independent of male nutrient intake. We tested whether females also have preferences based on variation in male courtship song, and whether the structure of male courtship song varies with nutrient intake. First, we reexamined female preference for calling song chirp rate. Then, we examined: (1) female preference based on courtship song chirp rate; (2) the relative importance of calling song and courtship song chirp rate; (3) the nutrition dependence of courtship song chirp rate; and (4) the correlation between calling song and courtship song chirp rate. As reported previously, females preferred higher calling song chirp rates, and in addition, preferred higher courtship song chirp rates. Females were more likely to switch from a speaker broadcasting more attractive calling song to a speaker broadcasting less attractive calling song when the attractive calling song was associated with an unattractive courtship song than when it was associated with an attractive courtship song. Preferences based on courtship song may thus cause females to alter the choices that they made based on calling song. Males that received greater nutrients did not produce higher courtship song chirp rates. There was no correlation between calling song and courtship song chirp rate. As a result, the two traits may provide information to females about different aspects of male quality. Copyright 2000 The Association for the Study of Animal Behaviour.     

Nocturnal and diurnal singing activity in the nightingale: correlations with mating status and breeding cycle

This study on the nightingale, Luscinia megarhynchos, is the first to examine both nocturnal and diurnal singing activity of mated and unmated males throughout a species' entire breeding cycle. Nocturnal song was sung mostly by unmated males. After pair formation, males ceased nocturnal singing and resumed it if their mate deserted. These results strongly suggest that nocturnal song of unmated males functions to attract a mate. Diurnal singing activity before females settled was low and did not predict future mating status. However, unmated males showed a continuous increase in diurnal singing activity until the end of the breeding cycle, but diurnal singing activity of mated males decreased after the egg-laying period. Mated males resumed nocturnal song for, on average, 3 nights during egg laying by their mates. This second period of nocturnal song coincided with the peak of diurnal singing activity. Such a high male singing effort during egg laying might allow the female to adjust her reproductive effort to male quality, deter rival males (e.g. through honest announcement of the female's fertility) or attract females for extrapair copulations. Copyright 2002 The Association for the Study of Animal Behaviour. Published by Elsevier Science Ltd. All rights reserved.     

Increased syllable rate during aggressive singing in a bird with complex and fast song

Sexual signals can comprise traits with multiple functions, and species with extreme phenotypes offer an opportunity to link function with signal evolution. This is the case in the serin Serinus serinus, a songbird with extremely fast syllable rate compared to related finches, and high sound frequency for its body size. Previous work on receiver responses showed that playback of artificially increased syllable rate is avoided and inhibits vocal responses, suggesting it is perceived as aggressive, while, on the contrary, higher sound frequency appears preferred by females. We tested whether senders also change these traits during aggressive singing, with a field playback experiment. Serin males responding aggressively by approaching the playback loudspeaker also increased syllable rate, while males responding less aggressively did not change syllable rate. Together with work on receiver responses, this suggests that aggressive signalling may have been an important selective pressure for the evolution of extremely fast syllable rate in this species. It is noteworthy that aggressive male serins still increase syllable rate, despite of their already elevated natural syllable rate. We found no changes in sound frequency when singing aggressively, which agrees with previous work that instead showed a female preference for high song frequency. We conclude that the evolution of extreme traits in serin song is best explained by multiple functions.     

Antiphonal duetting and sex hormones in the tropical bush shrike Laniarius funebris (HARTLAUB).

We investigated changes in antiphonal duetting with phases of reproduction and circulating levels of luteinizing hormone, testosterone, and estradiol in slate-colored boubous (Laniarius funebris) breeding in aviaries. Frequency of overall male singing did not vary with reproductive phase while frequencies of female singing and female vocal responses to male song were reduced during incubation and feeding of nestlings. This resulted in significant changes in frequency of duetting. Males sang the sexual song type M1 more often during courtship and nest building than during the nestlings phase. Their territorial song types M2 and M4 did not vary with breeding phase. Females were less responsive to M1 during incubation and to M2 during the nest building and nestlings than during the courtship phase. Plasma levels of luteinizing hormone and testosterone increased in males from the prebreeding to the courtship phase. While testosterone decreased already during nest building and remained low during subsequent phases of reproduction, luteinizing hormone decreased during incubation and feeding of nestlings. Female luteinizing hormone levels were highest during nest building. Female estradiol levels decreased from nest building to incubation and increased again during subsequent nest building. Female testosterone levels were low but not basal and did not vary with phase. Neither the overall male and female singing frequencies nor the frequencies of male song types were correlated with hormonal state. However, female participation in territorial duets M4 correlated positively with their testosterone levels. It is suggested that in this monogamous, duetting species with prolonged pairbonds behavioral cues between the mates are more important than the hormonal state in control of male and female singing.     

The Effect of Male Competition on the Courtship Song of Drosophila melanogaster

The courtship song of Drosophila has been extensively used as a model system for studies of sexual selection and species recognition. Traditionally, the courtship song has been recorded from males placed individually with a female. However, under natural conditions females are exposed to multiple courting males, and the effect of their joint signal on mate recognition by the female is not yet understood. Here, we recorded the courtship behavior of D. melanogaster males singing either individually to a female lpar;1:1) or in the presence of an additional male lpar;2:1). We compared the structure of the male song in the two experimental designs. Our results show that when two males courted a female their songs could overlap each other. Males produced a significantly different signal in the presence of competition; the duration of each song component was significantly shorter and the rate of singing was markedly lower. The present study demonstrates that male competition can dramatically alter the acoustic signal detected by the female.     

Der Gesang des weiblichen Kanarienvogels

Summary Isolated female canaries show song activity after the end of the reproductive period. Their repertoire is similar to those of themales. However, the utterances have a much higher range of variability in females than in males. Measurements of sex steroids give no support that the song controlling brain centres are short term activated by an increased titer of testosterone. Singing females show that the neural mechanisms for singing are differentiated in females, although singing is not practised during ontogeny.     

Variation in social relationships relates to song preferences and EGR1 expression in a female songbird

Social experiences can profoundly shape social behavior and the underlying neural circuits. Across species, the formation of enduring social relationships is associated with both neural and behavioral changes. However, it remains unclear how longer‐term relationships between individuals influence brain and behavior. Here, we investigated how variation in social relationships relates to variation in female preferences for and neural responses to song in a pair‐bonding songbird. We assessed variation in the interactions between individuals in male‐female zebra finch pairs and found that female preferences for their mate's song were correlated with the degree of affiliation and amount of socially modulated singing, but not with the frequency of aggressive interactions. Moreover, variation in measures of pair quality and preference correlated with variation in the song‐induced expression of EGR1, an immediate early gene related to neural activity and plasticity, in brain regions important for auditory processing and social behavior. For example, females with weaker preferences for their mate's song had greater EGR1 expression in the nucleus Taeniae, the avian homologue of the mammalian medial amygdala, in response to playback of their mate's courtship song. Our data indicate that the quality of social interactions within pairs relates to variation in song preferences and neural responses to ethologically relevant stimuli and lend insight into neural circuits sensitive to social information. © 2016 Wiley Periodicals, Inc. Develop Neurobiol 76: 1029–1040, 2016     

Song Choice Is Modulated by Female Movement in Drosophila Males

Mate selection is critical to ensuring the survival of a species. In the fruit fly, Drosophila melanogaster, genetic and anatomical studies have focused on mate recognition and courtship initiation for decades. This model system has proven to be highly amenable for the study of neural systems controlling the decision making process. However, much less is known about how courtship quality is regulated in a temporally dynamic manner in males and how a female assesses male performance as she makes her decision of whether to accept copulation. Here, we report that the courting male dynamically adjusts the relative proportions of the song components, pulse song or sine song, by assessing female locomotion. Male flies deficient for olfaction failed to perform the locomotion-dependent song modulation, indicating that olfactory cues provide essential information regarding proximity to the target female. Olfactory mutant males also showed lower copulation success when paired with wild-type females, suggesting that the male’s ability to temporally control song significantly affects female mating receptivity. These results depict the consecutive inter-sex behavioral decisions, in which a male smells the close proximity of a female as an indication of her increased receptivity and accordingly coordinates his song choice, which then enhances the probability of his successful copulation.     

Different singing styles in mated and unmated Reed Buntings Emberiza schoeniclus

Ewin described two singing styles, "rapid" and "slow", in the Reed Bunting Emberiza schoeniclus. My research has demonstrated that the prevalence of song styles is related to male mating status. Songs of mated and unmated males differed in the interval structure, number of syllables per song, number of frequency modulations per syllable and number of introductory syllables. Since both singing styles are uttered at the same intensity, it is not likely that they are parts of a continuous range and it is justified to treat them as two different song categories. After losing their mate, males sang like unpaired males and therefore a learning effect can be excluded. Both singing styles were directed to females, the slow one probably only to the mate. The use of these singing styles as a means of territory or mate defence cannot be excluded, but, if so used, it would seem to be of minor significance.     

The Onset of Song Learning and Song Tutor Selection in Fledgling Zebra Finches

We investigated song development in the pre‐independent zebra finch (aged 15–35 d), a period when neural pathways for song learning and production are forming and social influences outside the family are limited. Expt 1 investigated the onset and the minimum duration of tutoring needed for song learning in fledglings. We found most begin to learn song from 25 d of age and need about 10 d contact with the father tutor to make accurate copies. This onset corresponds with major developments in the formation of the neural circuitry implicated in song acquisition. Subsong also begins on day 25 suggesting that the sensory and motor phases of song learning fully overlap in the zebra finch. Our findings support the hypothesis that the song circuitry is fully functional by 35 d of age and the sensitive phase for zebra finches extends therefore from about days 25–65. However, only the first 10 d of this period are necessary to learn a tutor's song with fair accuracy. Expt 2 investigated the role of the paternal bond, spatial proximity and mating status in a fledgling's choice of song tutor where the father was the sole parent. Young chose the father over single unrelated males (expt 2a) or unrelated males in company with their female partners (expt 2b). Given the close spatial proximity of both potential tutors to the fledglings it is probably the filial bond, established via paternal care that is the cause of this preference. Zebra finches sing the same song phrase in two contrasting contexts: female‐directed song during pre‐copulatory courtship and undirected song where no female or display is involved. In expt 3 we tested the song learning preference of pre‐independent young for two categories of non‐paternal tutors: those singing predominantly female‐directed song and those singing exclusively undirected song. There was a small, but significant, preference for fledgling zebra finches to copy songs from males that sang female‐directed song. This preference is consistent with the hypothesis that young males not only learn the acoustic features of their tutor's song but also the visual and dynamic movements that constitute the courtship display.