Experimental support for a model of birdsong production.

In this work we present an experimental validation of a recently proposed model for the production of birdsongs. We have previously observed that driving the model with simple functions of time, which represent tensions in vocal muscles, produces a wide variety of sounds resembling birdsongs. In this work we drive the model with functions whose time dependence comes from recordings of muscle activities and air sac pressure. We simultaneously recorded the birds' songs and compared them with the synthetic songs. The model produces recognizable songs. Beyond finding a qualitative agreement, we also test some predictions of the model concerning the relative levels of activity in the gating muscles at the beginning and end of a syllable.

Parvalbumin-positive projection neurons characterise the vocal premotor pathway in male, but not female, zebra finches.

Parvalbumin (PV) and calbindin (CB) immunoreactivities were assessed in nucleus robustus archistriatalis (RA) of male and female zebra finches, together with retrograde labelling of RA neurons. The results of double and triple labelling experiments suggested that, in males, moderately and faintly PV-positive neurons were projection neurons, but that all intensely PV-positive cells were not. The latter, which are presumably interneurons, were also intensely CB-positive, and may correspond to the GABAergic inhibitory interneurons identified by others. In addition, the complete RA pathway and its terminal fields in the respiratory-vocal nuclei of the brainstem were strongly PV-positive. In female zebra finches, which do not sing, no evidence was found that PV-positive RA cells were projection neurons, yet the pattern of projections of RA neurons, as determined by anterograde transport of biotinylated dextran amine, was very similar to that of RA in males. Moreover, in females, RA neurons retrogradely labelled from injections of cholera toxin B-chain into the tracheosyringeal nucleus (XIIts) were abundant and included, in the lateral part of the nucleus, a population of cells that were as large as those in the male RA. Parvalbumin immunoreactivity was also present in RA and its projections in males of several other songbird species (northern cardinal, brown headed cowbird, canary) and in the female cardinal, which sings to some extent, but the labelling was not as intense as that in male zebra finches.

Neural pathways for bilateral vocal control in songbirds.

Ipsilateral and contralateral projections of nucleus robustus archistriatalis (RA), a telencephalic vocal premotor nucleus, to respiratory-vocal nuclei in the brainstem were defined in adult male Wasserschlager canaries, grey catbirds, and zebra finches, three songbird species that appear to differ in the degree of lateralized syringeal dominance. In all three species, ipsilateral projections of RA to the medulla included the tracheosyringeal part of the hypoglossal nucleus (XIIts), that innervates the syrinx, the bird's vocal organ, the suprahypoglossal area (SH), and two respiratory-related nuclei, retroambigualis (RAm) and parambigualis (PAm; Reinke and Wild [1998] J Comp Neurol 391:147-163). Projections of RA to the contralateral XIIts, SH and RAm, were substantial in canaries, which use the left side of the syrinx predominantly during singing; less pronounced in catbirds, which have no lateral dominance for song control; and least pronounced in zebra finches, in which there is a right-sided dominance for song control. There were no obvious differences in the number of crossed projections in birds injected in the left or right RA. Local sources of inputs to XIIts and RAm were defined anatomically in zebra finches and canaries. RAm, including neurons in close proximity to XIIts, was found to project to XIIts and the suprahypoglossal area bilaterally but predominantly ipsilaterally. RAm also had reciprocal connections with its contralateral homologue. These results suggest a pattern of connections between premotor and motor respiratory-vocal nuclei that may be involved in bilateral control of vocal output at medullary levels, a control that involves a high degree of coordination between vocal and respiratory structures on both sides of the body.

Sensitive period for sensorimotor integration during vocal motor learning.

Sensory experience during sensitive periods in development may direct the organization of neural substrates, thereby permanently influencing subsequent adult behavior. We report a sensitive period during the imitative motor learning phase of sensorimotor integration in birdsong development. By temporarily and reversibly blocking efference to the vocal muscles, we disrupted vocal motor practice during selected stages of song development. Motor disruption during prolonged periods early in development, which allows recovery of vocal control prior to the onset of adult song, has no effect on adult song production. However, song disruption late in development, during the emergence of adult song, results in permanent motor defects in adult song production. These results reveal a decreased ability to compensate for interference with motor function when disturbances occur during the terminal stage of vocal motor development. Temporary disruption of syringeal motor control in adults does not produce permanent changes in song production. Permanent vocal aberrations in juveniles are evident exclusively in learned song elements rather than nonlearned calls, suggesting that the sensitive period is associated with motor learning.

Bilaterally symmetrical respiratory activity during lateralized birdsong.

We investigated whether activity of expiratory muscles reflects lateralized activity of the vocal organ during production of birdsong. Respiration and syringeal motor activity were assessed in brown thrashers by monitoring bilateral airflow and subsyringeal air sac pressure, together with the electromyographic activity of expiratory abdominal muscles and vocal output. Activity of expiratory muscles was always present on both sides, regardless of whether song was produced bilaterally or on only one side of the syrinx. The average amplitude of expiratory EMG of one side does not change significantly, even if that side is silent during phonation. The temporal pattern of the electromyogram (EMG) was similar on both sides. Bilateral bursts of EMG activity on both sides accompanied changes in the rate of syringeal airflow, even when these flow fluctuations were generated only by one side of the syrinx. Motor commands to the respiratory muscles therefore appear to be bilaterally distributed, in contrast to the lateralized motor control of the syrinx.

A bird's own song contributes to conspecific song perception.

We investigated the role of developmental vocal experience in adult song perception by muting juvenile male zebra finches prior to song development and testing their behavioral responses to song playback as adults. Birds were raised in a normal social and acoustic environment. Non-treated sibling control birds demonstrated statistically significant phonotactic preferences for particular conspecific familiar or novel songs. Muted birds responded to playbacks at chance levels, showing no preferences for individual conspecific songs. These results suggest that the acquisition of a bird's own song may contribute to the perceptual processing, recognition, or discrimination of different conspecific songs.

The neuromuscular control of birdsong.

Birdsong requires complex learned motor skills involving the coordination of respiratory, vocal organ and craniomandibular muscle groups. Recent studies have added to our understanding of how these vocal subsystems function and interact during song production. The respiratory rhythm determines the temporal pattern of song. Sound is produced during expiration and each syllable is typically followed by a small inspiration, except at the highest syllable repetition rates when a pattern of pulsatile expiration is used. Both expiration and inspiration are active processes. The oscine vocal organ, the syrinx, contains two separate sound sources at the cranial end of each bronchus, each with independent motor control. Dorsal syringeal muscles regulate the timing of phonation by adducting the sound-generating labia into the air stream. Ventral syringeal muscles have an important role in determining the fundamental frequency of the sound. Different species use the two sides of their vocal organ in different ways to achieve the particular acoustic properties of their song. Reversible paralysis of the vocal organ during song learning in young birds reveals that motor practice is particularly important in late plastic song around the time of song crystallization in order for normal adult song to develop. Even in adult crystallized song, expiratory muscles use sensory feedback to make compensatory adjustments to perturbations of respiratory pressure. The stereotyped beak movements that accompany song appear to have a role in suppressing harmonics, particularly at low frequencies.

Inspiratory muscle activity during bird song.

The apparently continuous flow of bird song is in reality punctuated by brief periods of silence during which there are short inspirations called minibreaths. To determine whether these minibreaths are accompanied, and thus perhaps caused, by activity in inspiratory muscles, electromyographic (EMG) activity was recorded in M. scalenus in zebra finches and in M. scalenus and Mm. levatores costarum in cowbirds, together with EMGs from the abdominal expiratory muscles, air sac pressure and tracheal airflow. EMG activity in Mm. scalenus and levatores costarum consistently preceded the onset of negative air sac pressure by approximately 11 ms during both quiet respiration and singing in both species. The electrical activity of these two muscles was very similar. Compared with during quiet respiration, the amplitude of inspiratory muscle EMG during singing was increased between five- and 12-fold and its duration was decreased from >200 ms to on average 41 ms during minibreaths, again for both species, but inspiratory muscle activity did not overlap with that of the expiratory muscles. Thus, there was no indication that the inspiratory muscles acted either to shorten the duration of expiration or to reduce the expiratory effort as might occur if both expiratory and inspiratory muscles were simultaneously active. Inspiratory and expiratory muscle activities were highly stereotyped during song to the extent that together, they defined the temporal pattern of the songs and song types of individual birds.

Peripheral control and lateralization of birdsong.

Recent studies on several species of oscine songbirds show that they achieve their varied vocal performances through coordinated activity of respiratory, syringeal, and other vocal tract muscles in ways that take maximum advantage of the acoustic flexibility made possible by the presence of two independently controlled sound sources in their bipartite syrinx (vocal organ). During song, special motor programs to respiratory muscles alter the pattern of ventilation to maintain the supply of respiratory air and oxygen to permit songs of long duration, high syllable repetition rates, or maximum spectral complexity. Each side of the syrinx receives its own motor program that, together with that sent to respiratory muscles, determines the acoustic properties of the ipsilaterally produced sound. The acoustic expression of these bilaterally distinct, phonetic motor patterns depends on the action of dorsal syringeal adductor muscles that, by opening or closing the ipsilateral side of the syrinx to airflow, determine the amount each side contributes to song. The syringeally generated sound is further modified by muscles that control the shape of the vocal tract. Different species have adopted different motor strategies that use the left and right sides of the syrinx in patterns of unilateral, bilateral, alternating, or sequential phonation to achieve the differing temporal and spectral characteristics of their songs. As a result, the degree of song lateralization probably varies between species to form a continuum from unilateral dominance to bilateral equality.

Role of syringeal muscles in controlling the phonology of bird song.

1. The contribution of syringeal muscles to controlling the phonology of song was studied by recording bilateral airflow, subsyringeal air sac pressure, electromyograms (EMGs) of six syringeal muscles, and vocal output in spontaneously singing brown thrashers (Toxostoma rufum). 2. EMG activity in musculus syringealis ventralis (vS), the largest syringeal muscle, increases exponentially with the fundamental frequency of the ipsilaterally generated sound and closely parallels frequency modulation. 3. The EMG activity of other syringeal muscles is also positively correlated with sound frequency, but the amplitude of their EMGs changes only a small amount compared with variation in the amplitude of their EMGs correlated with changing syringeal resistance. The elevated activity in all syringeal muscles during high-frequency sounds may reflect an increased need for structural stability during the strong contractions of the largest syringeal muscle (vS). 4. Several syringeal mechanisms are used to generate amplitude modulation (AM). The most common of these involves modulating the rate of syringeal airflow, through activity by adductor (m. syringealis dorsalis and m. tracheobronchialis dorsalis) and abductor (m. tracheobronchialis ventralis) muscles, which change syringeal resistance, switch sound production from one side of the syrinx to the other, or produce rapid oscillatory flow changes. Variation in the phase relationship between AM and EMG bursts during oscillatory airflow suggests complex biomechanical interaction between antagonistic muscles. 5. AM can also arise from acoustic interactions of two independently generated sounds (beat notes) including cross talk signals between the two syringeal halves. In this latter mechanism, sound generated on one side radiates slightly out of phase with the source from the contralateral side, resulting in lateralized AM generation.

Motor stereotypy and diversity in songs of mimic thrushes.

The relationship between the motor and acoustic similarity of song was examined in brown thrashers (Toxostoma rufum) and grey catbirds (Dumetella carolinensis) (family Mimidae), which have very large song repertoires and sometimes mimic other species. Motor similarity was assessed by cross correlation of syringeal airflows and air sac pressures that accompany sound production. Although most syllables were sung only once in the song analyzed, some were repeated, either immediately forming a couplet, or after a period of intervening song, as a distant repetition. Both couplets and distant repetitions are produced by distinctive, stereotyped motor patterns. Their motor similarity does not decrease as the time interval between repetitions increases, suggesting that repeated syllables are stored in memory as fixed motor programs. The acoustic similarity between nonrepeated syllables, as indicated by correlation of their spectrograms, has a significant positive correlation with their motor similarity. This correlation is weak, however, suggesting that there is no simple linear relationship between motor action and acoustic output and that similar sounds may sometimes be produced by different motor mechanisms. When compared without regard to the sequence in which they are sung, syllables paired for maximum spectral similarity form a continuum with repeated syllables in terms of their acoustic and motor similarity. The prominence of couplets in the "syntax" of normal song is enhanced by the dissimilarity of successive nonrepeated syllables that make up the remainder of the song.

Role of syringeal muscles in gating airflow and sound production in singing brown thrashers.

1. The role of syringeal muscles in song production, particularly in regulating airflow through the syrinx, was studied in singing brown thrashers (Toxostoma rufum). In nine individuals, muscle activity was recorded electromyographically together with bilateral syringeal airflow, subsyringeal air sac pressure, and vocal output. 2. Dorsal muscles, m. syringealis dorsalis (dS) and m. tracheolateral dorsalis (dTB), are consistently activated during ipsilateral closing of the syrinx or increasing syringeal resistance, suggesting that their main role is adduction. This interpretation is supported by the motor patterns accompanying syllables with rapid oscillations in the rate of airflow. Bursts of electrical activity (2-10 ms) in dorsal muscles are precisely synchronized with decreasing airflow. 3. Electrical activity in m. tracheobronchialis ventralis (vTB) and m. tracheolateralis (TL) is associated with active abduction. An important contribution of vTB is to open the syringeal lumen for short inspirations in between syllables. In syllables with oscillatory flow modulations, vTB bursts show variable alignment with the phase of increasing flow. From this and activity during other syllables, it appears that, during phonation, vTB activity fine tunes the syringeal configuration, which is set by action of the dorsal muscles into a partially constricted state. 4. Activity in the ventral portion of TL, an extrinsic muscle, is strikingly similar to that of vTB, an intrinsic muscle, suggesting that the two muscles have a similar functional role. This supports the notion that intrinsic syringeal muscles of songbirds evolved from extrinsic muscles of nonpasserines. 5. M. syringealis ventralis (vS) does not appear to contribute directly to gating of airflow. Its activity is not consistently correlated with active changes in syringeal resistance. 6. Activity in m. sternotrachealis (ST) is most prominent during rapid changes in the rate of airflow or when switching between expiratory and inspiratory flow, suggesting a role in stabilizing the syringeal framework.

Variable asymmetry and resonance in the avian vocal tract: a structural basis for individually distinct vocalizations.

The social vocalizations of the oilbird (Steatornis caripensis) frequently have their acoustic energy concentrated into 3 prominent formants which appear to arise from the filter properties of their asymmetrical vocal tract with its bronchial syrinx. The frequency of the second and third formants approximate the predicted fundamental resonances of the unequal left and right cranial portions of each primary bronchus, respectively. Reversibly plugging either bronchus eliminates the corresponding formant. The first formant may arise in the trachea. The degree of vocal tract asymmetry varies between individuals, endowing them with different formant frequencies and providing potential acoustic cues by which individuals of this nocturnal, cave dwelling species may recognize each other in their dark, crowded colonies.

Lateralization and motor stereotypy of song production in the brown-headed cowbird.

Song production in adult brown-headed cowbirds (Molothrus ater ater) is lateralized, with a slight right syringeal dominance. The left side of the syrinx produces low-frequency (200-2000 Hz) notes within the introductory note clusters, while the right side produces the higher-frequency (1500-6000 Hz) introductory notes, the interphrase unit (10-12 kHz), and the final high-frequency whistle (5-13 kHz). Cross-correlation analyses reveal that individual cowbirds produce each of their four to seven song types with a distinct stereotyped motor pattern--as judged by the patterns of syringeal airflow and subsyringeal pressure. The acoustic differences across song types are reflected in the differences in the bronchial airflow and air sac pressure patterns associated with song production. These motor differences are particularly striking within the second and third introductory note clusters where there is a rapid switching back and forth between the two sides of the syrinx in the production of notes. These motor skills may be especially important in producing behaviorally effective song.

Motor dynamics of song production by mimic thrushes.

In brown thrashers (Toxostoma rufum) and grey catbirds (Dumetella carolinensis) neither side of the syrinx has a consistently dominant role in song production. During song, the two sides operate independently, but in close cooperation with each other and with the respiratory muscles which are capable of adjusting expiratory effort to maintain a constant rate of syringeal airflow despite sudden changes in syringeal resistance. Phonation is frequently switched from one side of the syrinx to the other, both between syllables and within a syllable. When both sides of the syrinx produce sound simultaneously, their respective contributions are seldom harmonically related. The resulting "two-voice" syllables sometimes contain difference tones with prominent sinusoidal amplitude modulation (AM). Rarely, both sides simultaneously produce the same sound. In general, however, the frequency range of sound contributed by the right syrinx is higher than that of the left syrinx. The right syrinx is also primarily responsible for producing a rapid cyclical amplitude modulation which is a characteristic feature of some syllables. This kind of AM is generated by either repetitive brief bursts of sound from the right side that modulate the amplitude of a continuous sound arising on the left side or cyclically opening the right syrinx, allowing unmodulated expiratory air to bypass the phonating left side.

Lateralization of syringeal function during song production in the canary.

The canary (Serinus canaria) vocal organ, the syrinx, has two separate sound sources, one in the cranial end of each bronchus. Previous investigations of whether song syllables are produced unilaterally or bilaterally have provided two contradictory results, as one researcher suggested that almost all syllables are produced by the left side of the syrinx alone, whereas another researcher suggested that both sides contribute similarly to all syllables. Our experiments, which involved unilateral bronchus plugging followed later by denervation of the ipsilateral syringeal muscles, attempted to resolve this disagreement. The males with right bronchus plugs, singing on the left side of the syrinx alone, produced nearly normal songs, whereas the birds with left bronchus plugs, singing on the right side, sang quite poorly. Interpretation of these data is difficult because it is not clear how syringeal function would be affected if the airflow rate through the intact side is increased above normal, nor is it known if the bird can compensate for bronchus occlusion. Nonetheless, we suggest that in male canaries most syllables are normally sung by the left side alone, with some syllables being produced by the right side alone and some being sung by both sides together. Right nerve section had little effect on the right-bronchus-plugged males' ability to sing, but the repertoires of the left-plugged males were altered after left nerve section, indicating the possibility that signals carried by the left nerve exert an influence on the contralateral side.

The acoustic role of tracheal chambers and nasal cavities in the production of sonar pulses by the horseshoe bat, Rhinolophus hildebrandti.

The acoustic role of the enlarged, bony, nasal cavities and rigid tracheal chambers in the horseshoe bat, Rhinolophus hildebrandti (Fig. 2) was investigated by determining the effect of their selective filling on the nasally emitted sonar pulse and on the sound traveling backwards down the trachea. Normal sonar signals of this bat contain a long constant frequency component with most energy in the second harmonic at about 48 kHz. The fundamental is typically suppressed 20 to 30 dB below the level of the second harmonic (Fig. 1). None of the experimental manipulations described affected the frequency of the sonar signal fundamental. Filling the dorsal and both lateral tracheal chambers had little effect on the emitted vocalization, but caused the level of the fundamental component in the trachea to increase 15 to 19 dB in most bats (Table 2). When only the dorsal chamber or only the two lateral chambers were filled, the effect was less striking and more variable (Tables 3 and 4), suggesting that the tracheal fundamental is normally suppressed by acoustic interaction between these three cavities. Filling the enlarged dorsal nasal cavities had no effect on the tracheal sound. The effect of this treatment on the nasally emitted sonar pulse was inconsistent. Sometimes the fundamental increased 10 to 12 dB, other times the intensity of all harmonics decreased; in still other cases the second, third or fourth harmonic increased, but the fundamental remained unchanged (Tables 5, 6, and 7). When bats were forced to vocalize through the mouth, by sealing the nostrils, there was a prominent increase in the level of the emitted fundamental (10 to 21 dB) and in the fourth harmonic (6 to 17 dB). In one instance there was also a significant increase in the level of the third harmonic (Tables 8 and 9). The supraglottal tract thus filters the fundamental from the nasally emitted sonar signal, although the role of the inflated nasal cavities in this process is unclear. We conclude that a high glottal impedance acoustically isolates the subglottal from the supraglottal vocal tract. The tracheal chambers do not affect the emitted sonar signal, but may attenuate the fundamental in the trachea and prevent it from being reflected from the lungs back towards the cochlea. It may be important to prevent the reflected fundamental from stimulating the cochlea, via tissue conduction, along multiple indirect pathways which would temporally smear cochlear stimulation.(ABSTRACT TRUNCATED AT 400 WORDS)

The sound emission pattern and the acoustical role of the noseleaf in the echolocating bat, Carollia perspicillata.

Carollia perspicillata (Phyllostomidae) is a frugivorous bat that emits low-intensity, broadband, frequency-modulated echolocation pulses through nostrils surrounded by a noseleaf. The emission pattern of this bat is of interest because the ratio between the nostril spacing and the emitted wavelength varies during the pulse, causing complex interference patterns in the horizontal dimension. Sound pressures around the bat were measured using a movable microphone and were referenced to those at a stationary microphone positioned directly in front of the animal. Interference between the nostrils was confirmed by blocking one nostril, which eliminated sidelobes and minima in the emission pattern, and by comparison of real emission patterns with simple computer models. The positions of minima in the patterns indicate effective nostril spacings of over a half-wavelength. Displacement of the dorsal lancet of the noseleaf demonstrated that this structure directs sound in the vertical dimension.