Male Song Production Research Articles

Sexual signal loss, pleiotropy, and maintenance of a male reproductive polymorphism in crickets.

Pleiotropy between male signals and female preferences can facilitate evolution of sexual communication by maintaining coordination between the sexes. Alternatively, it can favor variation in the mating system, such as a reproductive polymorphism. It is unknown how common either of these scenarios is in nature. In Pacific field crickets (Teleogryllus oceanicus) on Kauai, Hawaii, a mutation (flatwing) that segregates as a single locus is responsible for the rapid loss of song production in males. We used outbred cricket colonies fixed for male wing morph to investigate whether homozygous flatwing and normal-wing (wild-type) females differ in responsiveness to male calling song and propensity to mate when paired with either a flatwing or normal-wing male in the presence or absence of courtship song. Flatwing females were less likely to mount a male than normal-wing females. Females of both genotypes showed a preference for normal-wing males and were more likely to mate in the presence of courtship song; normal-wing females were particularly likely to mate with song. Our results show that negative pleiotropy between obligate male silence and female mating behavior can constrain the evolution of sexual signal loss and contribute to the maintenance of a male reproductive polymorphism in the wild.

Frequency tuning and directional sensitivity of tympanal vibrations in the field cricket Gryllus bimaculatus.

Female field crickets use phonotaxis to locate males by their calling song. Male song production and female behavioural sensitivity form a pair of matched frequency filters, which in Gryllus bimaculatus are tuned to a frequency of about 4.7 kHz. Directional sensitivity is supported by an elaborate system of acoustic tracheae, which make the ears function as pressure difference receivers. As a result, phase differences between left and right sound inputs are transformed into vibration amplitude differences. Here we critically tested the hypothesis that acoustic properties of internal transmissions play a major role in tuning directional sensitivity to the calling song frequency, by measuring tympanal vibrations as a function of sound direction and frequency. Rather than sharp frequency tuning of directional sensitivity corresponding to the calling song, we found broad frequency tuning, with optima shifted to higher frequencies. These findings agree with predictions from a vector summation model for combining external and internal sounds. We show that the model provides robust directional sensitivity that is, however, broadly tuned with an optimum well above the calling song frequency. We therefore advocate that additional filtering, e.g. at a higher (neuronal) level, significantly contributes to frequency tuning of directional sensitivity.

Dominance-related seasonal song production is unrelated to circulating testosterone in a subtropical songbird

Circulating testosterone (T) is widely considered to play a key role in the production of sexual displays by male vertebrates. While numerous studies support a role for circulating T in promoting the production of song in male birds, this understanding is based primarily on evidence from seasonally breeding northern temperate species, leaving it unclear whether this mechanism generalizes to other regions of the world. Here we investigate whether variation in circulating levels of T can explain the marked within- and among-individual variation in male song performance observed in a subtropical population of the year-round territorial white-browed sparrow weaver (Plocepasser mahali mahali). Our findings reveal that both circulating T and male song production peaked at a similar time point, halfway through the population-level breeding season. However, while dominant males were more likely to sing and sang for longer than subordinate males, within-group paired comparisons revealed no dominance-related differences in circulating T. Moreover, comparisons both among and within individual dominant males revealed that song duration, syllable rate and proportion of time spent singing were all unrelated to circulating T. Together, our findings suggest that natural variation in male song production, at least in this population of white-browed sparrow weavers, is achieved principally through mechanisms other than variation in circulating T concentration. More widely, our results are in line with the view that male song production is not exclusively regulated by gonadally synthesized steroids.

Sexually dimorphic perineuronal net expression in the songbird

Sexual dimorphism of song behavior and its underlying brain architecture characterizes the vocal learning system of many songbirds (Nottebohm and Arnold [1] and Ball and MacDougall-Shackleton [2]). Females do not sing and their neural song control system is reduced compared to males. However, females do learn to discriminate male song during a sensitive period (Clayton [3] and Nelson et al. [4]). We reasoned that any song nuclei that might store developmental social learning in female zebra finches would be marked by extracellular perineuronal nets (PNNs), which limit plasticity at the ends of critical periods (Hockfield et al. [5] and Hensch [6]), stabilize learning (Wang and Fawcett [7]), and demarcate all song nuclei in male finches (Balmer et al. [8]). PNN expression patterns in four brain areas were compared in males and females: the Lateral Magnocellular Nucleus (LMAN) and a basal ganglia region that contains Area X in males, which together constitute an anterior cortical-basal ganglia pathway necessary for male song plasticity, and HVC and the Robust Nucleus of the Arcopallium (RA), which together form a posterior pathway that is critical for song production in males. We found a dichotomy of PNN expression in the female song system: the anterior cortical-basal ganglia pathway was highly enriched for PNNs and indistinguishable from males, whereas the posterior pathway was essentially devoid of PNNs. These data indicate that social signaling in the female song system may undergo a true critical period, similar to sensory systems, and that limiting this critical period selectively involves the song cortical-basal ganglia pathway.

Early-life stress affects the behavioural and neural response of female song sparrows to conspecific song

In songbirds, both song production and song preferences may be influenced by early-life experience. Early-life stress impairs development of the song-control brain regions and permanently affects male song production. However, few studies have examined the effects of early-life stress on female song preferences or the brains of female songbirds. We exposed female song sparrows, Melospiza melodia, to stressors (food restriction or exogenous corticosterone treatment) early in development. When birds were adults, we determined the effects of the stressors on: (1) the behavioural response to high-complexity versus low complexity-songs as well as to conspecific versus heterospecific songs; (2) the volume of song-control brain regions; and (3) the immediate early gene (Zenk) response in auditory forebrain regions following exposure to either conspecific or heterospecific song. We found no significant effect of developmental stress on the strength of the behavioural response to high- versus low-complexity song, but control females showed more selectivity in their behavioural response to conspecific versus heterospecific song when compared with food-restricted or corticosterone-treated birds. We observed no effect of either stressor on volume of the song-control nuclei. Control females exposed to conspecific song had more Zenk-immunoreactive cells in the auditory brain regions than control females exposed to heterospecific song. In contrast, food-restricted and corticosterone-treated females did not have more Zenk-immunoreactive cells after exposure to conspecific song. These results show that stressors known to affect male song production might also affect neural processing of song by females, and their subsequent behavioural response to song.

HVC lesions modify immediate early gene expression in auditory forebrain regions of femalesongbirds

It is well established that auditory forebrain regions of oscine birds are essential for the encoding of species-typical songs and are, therefore, vital for recognition of song during sociosexual interactions. Regions such as the caudal medial nidopallium (NCM) and the caudal medial mesopallium (CMM) are involved in perceptual processing of song and the formation of auditory memories. There is an additional telencephalic nucleus, however, that has also been implicated in species recognition. This nucleus is HVC, a prominent nucleus that sits at the apex of the song system, and is well known for its critical role in song learning and song production in male songbirds. Here, we explore the functional relationship between auditory forebrain regions (i.e., NCM and CMM) and HVC in female canaries (Serinus canaria). We lesion HVC and examine immediate early gene responses to conspecific song presentation within CMM and NCM to explore whether HVC can modulate auditory responses within these forebrain regions. Our results reveal robust deficits in ZENK-ir in CMM and NCM of HVC-lesioned females when compared with control- and sham-lesioned females, indicating that functional connections exists between HVC and NCM/CMM. Although these connected regions have been implicated in song learning and production in males, they likely serve distinct functions in female songbirds that face the task of song recognition rather than song production. Identifying functional connections between HVC and auditory regions involved in song perception is an essential step toward developing a comprehensive understanding of the neural basis of song recognition.

Vocally correlated seasonal auditory variation in the house sparrow (Passer domesticus)

Songbirds exhibit seasonal plasticity in a broad variety of behavioral and morphological traits associated with reproduction. Changes in song production are well described while changes in song reception are not. In the present study, we test for seasonal variation in auditory processing of the house sparrow (Passer domesticus L.) using auditory brainstem responses (ABRs) to tone bursts. We measured amplitude and latency of the first ABR peak in spring, summer and autumn at stimulus frequencies from 0.8 to 6.4 kHz and intensity levels from 24 to 80 dB SPL. ABR thresholds were determined at each frequency using cross-correlation. Amplitude was greater in spring than in autumn at frequencies from 3.2 to 6.4 kHz whereas latency and thresholds exhibited no seasonal variation. The results indicate an increase in the number or temporal synchrony of responses from peripheral auditory neurons during the early breeding season. Changes in peripheral auditory processing may enhance temporal coding of the fine structure and envelope of song; thereby, improving assessment of encoded information in both sexes (e.g. individual identity and dominance status) and auditory feedback during song production in males. Peripheral auditory changes may be mediated by reproductive hormones, and could involve changes in hair cell density on the basilar papilla. Our results suggest that peripheral auditory processing of songbirds changes seasonally in parallel with other behavioral and morphological traits, such as song production.

Subcellular compartmentalization of aromatase is sexually dimorphic in the adult zebra finch brain

The vertebrate brain is a source of estrogen (E) via the expression of aromatase (E-synthase). In the zebra finch (Taeniopygia guttata), despite documented dimorphisms in E-action, no differences are detectable in circulating E, or the neural levels of aromatase transcription, activity, or somal protein expression. Studies of aromatase expression at the light- and electron-microscope levels reveal greater numbers of fibers and presynaptic boutons in adult males relative to females. We assayed aromatase activity and content in synaptosomes and microsomes from the anterior [containing lMAN and Area X (males)] and posterior telencephalon (containing HVC and RA) of adult birds. In contrast to non-song birds and mammals, both cell fractions contain abundant aromatase measurable in terms of activity (enzyme assays) and content (Western blots) with minimal enrichment in microsomes. From brain homogenates of identical concentration, aromatase activity was higher in the synaptosomal relative to the microsomal fraction, in males relative to females, and in the posterior compared to anterior telencephalon. These effects were driven by high levels of synaptosomal aromatase in the male posterior telencephalon. These data suggest that males possess more aromatase per presynaptic bouton, or a greater number of aromatase-containing presynaptic boutons than females in the posterior telencephalon. Further, the present report reveals synaptic aromatization as a considerable source of E in the zebra finch brain, and supports the idea that telencephalic synapses in and around the adult male song production nuclei may be exposed to higher levels of E compared to the female brain.

Immediate early gene activity in song control nuclei and brain areas regulating motivation relates positively to singing behavior during, but not outside of, a breeding context

In some species, such as songbirds, much is known about how the brain regulates vocal learning, production, and perception. What remains a mystery is what regulates the motivation to communicate. European starlings (Sturnus vulgaris) sing throughout most of the year, but the social and environmental factors that motivate singing behavior differ seasonally. Male song is highly sexually motivated during, but not outside of, the breeding season. Brain areas outside the song control system, such as the medial preoptic nucleus (POM) and ventral tegmental area (VTA), have been implicated in regulating sexually motivated behaviors in birds, including song. The present study was designed to explore whether these regions, as well as three song control nuclei [area X, the high vocal center (HVC), and the robust nucleus of the arcopallium (RA)], might be involved differentially in song produced within compared to outside of a breeding context. We recorded the behavioral responses of breeding and nonbreeding condition male starlings to the introduction of a female conspecific. Males did not show context-dependent differences in the overall amount of song sung. However, immunocytochemistry for the protein product of the immediate early gene cFOS revealed a positive linear relationship between the total amount of songs sung and number of cFOS-labeled cells in POM, VTA, HVC, and RA for birds singing during, but not outside of, a breeding context. These results suggest that these regions differentially regulate male song production depending on reproductive context. Overall the data support the hypothesis that the POM and VTA interact with the song control system, specifically HVC and RA, to regulate sexually motivated vocal communication in songbirds.

Adult female and male zebra finches show distinct patterns of spine deficits in an auditory area and in the song system when reared without exposure to normal adult song

Male songbirds typically require exposure to normal adult conspecific song during development in order to learn a normal song of their own. Females require exposure to conspecific song during development in order to select high-quality, learned song over the incomplete song produced by males reared in isolation. Altering males' opportunity for song learning during development affects the neuroanatomy of brain regions involved in song production (the song system), but in females the neural effects of song learning are unknown. We raised male and female zebra finches (Taeniopygia guttata) with differing amounts of exposure to singing males during development. At 120 days, we Golgi-stained their brains and measured the frequency of dendritic spines in brain areas used in song perception or production. We found that females reared with little or no exposure to song have 31% fewer dendritic spines per unit length of dendrite in caudomedial nidopallium (NCM), a brain area activated by song perception, compared to control females. The deprived females had small deficits in the frequency of spines in HVC, a region activated by song production in males. Males with limited exposure to song had a 24% lower spine density in HVC than controls but only a 10% lower density in NCM. These data support the hypothesis that NCM is important in auditory learning, while HVC is involved in sensorimotor learning, and that these capacities are differentially emphasized in the two sexes.

Singing as a handicap: the effects of food availability and weather on song output in the Australian reed warbler Acrocephalus australis

Bird song is generally regarded as a sexually selected trait, and may represent a reliable handicap signal under at least certain conditions. Females may use the degree of male song production as a reliable cue to male condition or territory quality. We investigated the effect of supplementary feeding on song output in the migratory Australian reed warbler Acrocephalus australis. We experimentally increased the food availability on alternate days, and recorded several weather variables. We measured song rate and song length independently. Supplementary fed birds sang more on feeding days than on non‐feeding days, while control birds did not show this effect. Song output was not significantly associated with any of the weather variables examined. Our results indicate that singing has the potential to serve as a reliable handicap signal to territorial food availability irrespective of the prevailing weather conditions. We discuss the role of energetic constraints and behavioural flexibility on the signaling function of song.

Evidence for Opioid Involvement in the Regulation of Song Production in Male European Starlings (Sturnus vulgaris).

Many social animals vocalize at high rates, suggesting that vocal communication is highly motivated and rewarding. In songbirds, much is known about the neural control of vocal behavior; however, little is known about neurobiological mechanisms regulating the motivation to communicate. This study examined a possible role for opioid neuropeptides in motivation and reward associated with song production in male European starlings (Sturnus vulgaris). Peripheral opioid blockade facilitated male song production. Furthermore, methionine-enkephalin immunolabeled fiber densities within brain regions in which opioids are known to regulate motivation and reward (i.e., the medial preoptic nucleus and ventral tegmental area) related positively to male song production. These data suggest that song production might be regulated by opioid activity within motivation and reward neural systems.

TrkB-like immunoreactivity in the song system of developing zebra finches

The neural song system in zebra finches develops for approximately the first 2 months after hatching. During that time, male-biased sexual dimorphisms emerge in the volume of song control nuclei as well as in the number and size of neurons within them. Brain derived neurotrophic factor (BDNF) has been documented in song control nuclei at various stages of development. Its high affinity receptor (tyrosine kinase B; trkB) is also in the song system, at least at around 1 month of age. The present study was designed to more completely describe the timing and potential location of BDNF action by investigating trkB expression during sexual differentiation of the song control nuclei. The pattern of immunoreactivity to a trkB antibody was examined in male and female zebra finches at post-hatching days 3–60. Labeling in somata and neuropil appeared to define the telencephalic components of the motor pathway (high vocal center and robust nucleus of the archistriatum) for song production in males from days 30 to 60, and in females on days 45 and 60 (high vocal center). These results are consistent with the hypothesis that the receptor, and its ligand BDNF, play a role in processes related to song learning in both sexes, including perhaps the motor component exhibited by developing males.

Temperature coupling in cricket acoustic communication. II. Localization of temperature effects on song production and recognition networks in Gryllus firmus.

Acoustic communication in Gryllus firmus is temperature-coupled: temperature induces parallel changes in male calling song temporal pattern, and in female preference for song. Temperature effects on song production and recognition networks were localized by selectively warming head or thorax or both head and thorax of intact crickets, then eliciting aggression song production (males) or phonotaxis to synthetic calling song (females). Because male song is produced by a thoracic central pattern generator (CPG), and because head ganglia are necessary for female song recognition, measurements of female phonotaxis under such conditions may be used to test the following competing hypotheses about organization of the song recognition network: 1. A set of neurons homologous to the male song CPG exist in the female, and are used as a template that determines preferred values of song temporal parameters for song pattern recognition (the common neural elements hypothesis), and 2. temporal pattern preference is determined entirely within the head ganglia. Neither selective warming of the head nor of the thorax was effective in changing female song preference, but simultaneous warming of head and thorax shifted preference toward a faster song in most preparations, as did warming the whole animal by raising ambient temperature. These results suggest that phonotactic preference for song temporal pattern is plurisegmentally determined in field crickets. Selective warming experiments during aggression song production in males revealed that syllable period is influenced but not completely determined by thoracic temperature; head temperature is irrelevant. The song CPG appears to receive some rate-setting information from outside the thoracic central nervous system.

Genetic and Molecular Analysis of the Love Song Preferences of Drosophila Females

The courtship songs of male Drosophila have been studied at the behavioural, genetic and molecular levels. Less attention has been paid to the female's responses to these songs. Playback experiments are described which suggest that courtship songs are an important component of female mate choice. Some of the implications of the behavioural responses of hybrid females between D. melanogaster and D. simulans are examined in the light of theories concerning the mechanisms by which insect communication systems might evolve. The role of the period gene in both male song production and in female song reception is considered, and the neural regions in the female which may be important for song integration are briefly discussed.

Altered Perception of Species-Specific Song by Female Birds After Lesions of a Forebrain Nucleus

Female birds that do not normally sing possess brain nuclei associated with song production in males. To determine whether one song nucleus, the caudal nucleus of the ventral hyperstriatum (HVc), acts in conspecific song perception, courtship responses of female canaries to canary and white-crowned sparrow songs were compared before and after HVc lesions. Bilateral lesions of a portion of the HVc resulted in copulation solicitations to heterospecific song as well as conspecific song. Control females continued to respond only to conspecific song. This suggests that the HVc is critical for conspecific song perception in females. Because female canaries do not normally sing, neurons in female HVc must develop response selectivity by a mechanism different from that proposed for male birds in the motor theory of song perception.