Passive acoustic monitoring, development of disturbance calls and differentiation of disturbance and advertisement calls in the Argentine croaker Umbrina canosai (Sciaenidae).

Disturbance and advertisement calls of the Argentine croaker Umbrina canosai were recorded from coastal Uruguayan waters. Dissections indicate typical sciaenid extrinsic swimbladder muscles present exclusively in males. Disturbance calls were produced when captive U. canosai were startled, chased with a net or grabbed by the tail. Calls were unusual for sciaenids because each pulse consisted of multiple cycles. The number of cycles per pulse and dominant frequency did not change with U. canosai size, but pulse duration and interpulse interval increased. Advertisement calls were recorded from unseen choruses in the field and confirmed with captive individuals in a large tank. Advertisement calls were recorded throughout the known range of the species in Uruguay indicating a continuous belt of spawning populations. Tank recordings of the same individuals permitted explicit comparisons between the two calls. Advertisement call pulses averaged 2·4 more cycles (11·0-8·6) although pulses of both calls were basically similar as were durations and dominant frequencies. Pulse number, however, differed markedly, averaging 13·6 and 3·4 pulses for disturbance and advertisement calls respectively. Furthermore, disturbance calls were produced as a rapid series with an interpulse interval of 26-31 ms whereas advertisement call patterns were less stereotyped and ranged from <100 to 450 ms. Multicycle pulses distinguished U. canosai from other sympatric sciaenids.

Explosive development of pectoral muscle fibres in large juvenile blue catfish Ictalurus furcatus.

As part of an effort on scaling of pectoral spines and muscles, the basis for growth was examined in six pectoral muscles in juvenile blue catfish Ictalurus furcatus, the largest catfish in North America. Fibre number increases slowly in fish from 13.0 to 26.4 cm in total length, doubles by 27.0 cm and remains stable in larger individuals. Simultaneously, mean fibre diameter decreases by half, caused by the addition of new small fibres, before increasing non-linearly in larger fish. The orders of magnitude disparity between the size at hatching and the size of large adults may have selected for rapid muscle fibre addition at a threshold size.

Functional morphology of the sonic apparatus in Ophidion barbatum (Teleostei, Ophidiidae).

Most soniferous fishes producing sounds with their swimbladder utilize relatively simple mechanisms: contraction and relaxation of a unique pair of sonic muscles cause rapid movements of the swimbladder resulting in sound production. Here we describe the sonic mechanism for Ophidion barbatum, which includes three pairs of sonic muscles, highly transformed vertebral centra and ribs, a neural arch that pivots and a swimbladder whose anterior end is modified into a bony structure, the rocker bone. The ventral and intermediate muscles cause the rocker bone to swivel inward, compressing the swimbladder, and this action is antagonized by the dorsal muscle. Unlike other sonic systems in which the muscle contraction rate determines sound fundamental frequency, we hypothesize that slow contraction of these antagonistic muscles produces a series of cycles of swimbladder vibration.

Geographical variation in sound production in the anemonefish Amphiprion akallopisos.

Because of pelagic-larval dispersal, coral-reef fishes are distributed widely with minimal genetic differentiation between populations. Amphiprion akallopisos, a clownfish that uses sound production to defend its anemone territory, has a wide but disjunct distribution in the Indian Ocean. We compared sounds produced by these fishes from populations in Madagascar and Indonesia, a distance of 6500 km. Differentiation of agonistic calls into distinct types indicates a complexity not previously recorded in fishes' acoustic communication. Moreover, various acoustic parameters, including peak frequency, pulse duration, number of peaks per pulse, differed between the two populations. The geographic comparison is the first to demonstrate 'dialects' in a marine fish species, and these differences in sound parameters suggest genetic divergence between these two populations. These results highlight the possible approach for investigating the role of sounds in fish behaviour in reproductive divergence and speciation.

Acoustic communication in two freshwater gobies: ambient noise and short-range propagation in shallow streams.

Noise is an important theoretical constraint on the evolution of signal form and sensory performance. In order to determine environmental constraints on the communication of two freshwater gobies Padogobius martensii and Gobius nigricans, numerous noise spectra were measured from quiet areas and ones adjacent to waterfalls and rapids in two shallow stony streams. Propagation of goby sounds and waterfall noise was also measured. A quiet window around 100 Hz is present in many noise spectra from noisy locations. The window lies between two noise sources, a low-frequency one attributed to turbulence, and a high-frequency one (200-500 Hz) attributed to bubble noise from water breaking the surface. Ambient noise from a waterfall (frequencies below 1 kHz) attenuates as much as 30 dB between 1 and 2 m, after which values are variable without further attenuation (i.e., buried in the noise floor). Similarly, courtship sounds of P. martensii attenuate as much as 30 dB between 5 and 50 cm. Since gobies are known to court in noisy as well as quiet locations in these streams, their acoustic communication system (sounds and auditory system) must be able to cope with short-range propagation dictated by shallow depths and ambient noise in noisy locations.

Acoustic communication in two freshwater gobies: the relationship between ambient noise, hearing thresholds and sound spectrum.

Two freshwater gobies Padogobius martensii and Gobius nigricans live in shallow (5-70 cm) stony streams, and males of both species produce courtship sounds. A previous study demonstrated high noise levels near waterfalls, a quiet window in the noise around 100 Hz at noisy locations, and extremely short-range propagation of noise and goby signals. To investigate the relationship of this acoustic environment to communication, we determined audiograms for both species and measured parameters of courtship sounds produced in the streams. We also deflated the swimbladder in P. martensii to determine its effect on frequency utilization in sound production and hearing. Both species are maximally sensitive at 100 Hz and produce low-frequency sounds with main energy from 70 to 100-150 Hz. Swimbladder deflation does not affect auditory threshold or dominant frequency of courtship sounds and has no or minor effects on sound amplitude. Therefore, both species utilize frequencies for hearing and sound production that fall within the low-frequency quiet region, and the equivalent relationship between auditory sensitivity and maximum ambient noise levels in both species further suggests that ambient noise shapes hearing sensitivity.

Weakfish sonic muscle: influence of size, temperature and season.

The influence of temperature, size and season on the sounds produced by the sonic muscles of the weakfish Cynoscion regalis are categorized and used to formulate a hypothesis about the mechanism of sound generation by the sonic muscle and swimbladder. Sounds produced by male weakfish occur at the time and location of spawning and have been observed in courtship in captivity. Each call includes a series of 6-10 sound pulses, and each pulse expresses a damped, 2-3 cycle acoustic waveform generated by single simultaneous twitches of the bilateral sonic muscles. The sonic muscles triple in mass during the spawning season, and this hypertrophy is initiated by rising testosterone levels that trigger increases in myofibrillar and sarcoplasmic cross-sectional area of sonic muscle fibers. In response to increasing temperature, sound pressure level (SPL), dominant frequency and repetition rate increase, and pulse duration decreases. Likewise, SPL and pulse duration increase and dominant frequency decreases with fish size. Changes in acoustic parameters with fish size suggest the possibility that drumming sounds act as an 'honest' signal of male fitness during courtship. These parameters also correlate with seasonally increasing sonic muscle mass. We hypothesize that sonic muscle twitch duration rather than the resonant frequency of the swimbladder determines dominant frequency. The brief (3.5 ms), rapidly decaying acoustic pulses reflect a low-Q, broadly tuned resonator, suggesting that dominant frequency is determined by the forced response of the swimbladder to sonic muscle contractions. The changing dominant frequency with temperature in fish of the same size further suggests that frequency is not determined by the natural frequency of the bladder because temperature is unlikely to affect resonance. Finally, dominant frequency correlates with pulse duration (reflecting muscle twitch duration), and the inverse of the period of the second cycle of acoustic energy approximates the recorded frequency. This paper demonstrates for the first time that the dominant frequency of a fish sound produced by a single muscle twitch is apparently determined by the velocity of the muscle twitch rather than the natural frequency of the swimbladder.

Movement and sound generation by the toadfish swimbladder.

Although sound-producing (sonic) muscles attached to fish swimbladders are the fastest known vertebrate muscles, the functional requirement for such extreme speed has never been addressed. We measured movement of the swimbladder caused by sonic muscle stimulation in the oyster toadfish Opsanus tau and related it to major features of the sound waveform. The movement pattern is complex and produces sound inefficiently because the sides and bottom of the bladder move in opposite in and out directions, and both movement and sound decay rapidly. Sound amplitude is related to speed of swimbladder movement, and slow movements do not produce perceptible sound. Peak sound amplitude overlaps fundamental frequencies of the male's mating call because of muscle mechanics and not the natural frequency of the bladder. These findings suggest that rapid muscle speed evolved to generate sound from an inefficient highly damped system.

Variability in the role of the gasbladder in fish audition.

The teleost gasbladder is believed to aid in fish audition by transferring pressure components of incoming sound to the inner ears. This idea is primarily based on both anatomical observations of the mechanical connection between the gasbladder and the ear, followed by physiological experiments by various researchers. The gasbladder movement has been modeled mathematically as a pulsating bubble. This study is extending the previous work on fish with a physical coupling of the gasbladder and ear by investigating hearing in two species (the blue gourami Trichogaster trichopterus, and the oyster toadfish Opsanus tau) without a mechanical linkage. An otophysan specialist (the goldfish Carassius auratus) with mechanical coupling, is used as the control. Audiograms were obtained with acoustically evoked potentials (e.g., auditory brainstem response) from intact fish and from the same individuals with their gasbladders deflated. In blue gourami and oyster toadfish, removal of gas did not significantly change thresholds, and evoked potentials had similar waveforms. In goldfish thresholds increased by 33-55 dB (frequency dependent) after deflation, and major changes in evoked potentials were observed. These results suggest that the gasbladder may not serve an auditory enhancement function in teleost fishes that lack mechanical coupling between the gasbladder and the inner ear.

Effects of fish size and temperature on weakfish disturbance calls: implications for the mechanism of sound generation.

To categorize variation in disturbance calls of the weakfish Cynoscion regalis and to understand their generation, we recorded sounds produced by different-sized fish, and by similar-sized fish at different temperatures, as well as muscle electromyograms. Single, simultaneous twitches of the bilateral sonic muscles produce a single sound pulse consisting of a two- to three-cycle acoustic waveform. Typical disturbance calls at 18 degrees C consist of trains of 2-15 pulses with a sound pressure level (SPL) of 74 dB re 20 microPa at 10 cm, a peak frequency of 540 Hz, a repetition rate of 20 Hz and a pulse duration of 3.5 ms. The pulse duration suggests an incredibly short twitch time. Sound pressure level (SPL) and pulse duration increase and dominant frequency decreases in larger fish, whereas SPL, repetition rate and dominant frequency increase and pulse duration decreases with increasing temperature. The dominant frequency is inversely related to pulse duration and appears to be determined by the duration of muscle contraction. We suggest that the lower dominant frequency of larger fish is caused by a longer pulse (=longer muscle twitch) and not by the lower resonant frequency of a larger swimbladder.

Continuous adult development of multiple innervation in toadfish sonic muscle.

The sonic muscle of the oyster toadfish Opsanus tau produces unfused contractions at over 200 Hz for mating call production, requiring extreme muscle fiber synchronization. This multiply innervated muscle is sexually dimorphic and grows for life by fiber proliferation and hypertrophy. Previous descriptions of its multiple innervation did not consider fish size or sex. We examined neuromuscular junction (NMJ) development in adult fish of both sexes between 123 and 343 mm in total length (24.7790 g in mass). The NMJ was a tubelike trough that varied in length from 8 to 178 microm. Troughs were usually straight, although some consisted of consecutive ovals and some were branched. Median length of NMJs increased linearly with fish length (r2=.40; p=.002) from 58 to 75 microm. Modal lengths were mostly between 50 and 60 microm and did not increase ontogenetically, indicating that the median increase was caused by a greater number of large junctions in older fish. Median interval between NMJs (measured from the beginning of one junction to the next) ranged from 92 to 116 microm and did not vary with fish size (r2=.06; p=.285). Considering muscle fiber elongation, the data indicate an increase from 60 to 140 NMJs per fiber during fish growth. There were no sexual differences in NMJ length or spacing. In view of the slow conduction velocity of sonic muscle fibers, the addition of new NMJs and the relatively constant distance between them supports rapid and synchronized contraction necessary for sound production in both sexes.

Comparison of sarcoplasmic reticulum capabilities in toadfish (Opsanus tau) sonic muscle and rat fast twitch muscle.

The sonic muscle of the oyster toadfish, Opsanus tau, can produce unfused contractions at 300 Hz. Electron microscopy shows a great abundance of the Sarcoplasmic reticulum (SR) in this muscle, but no functional characterization of the capabilities of the SR has been reported. We measured the oxalate-supported Ca2+ uptake rate and capacities of homogenates of toadfish sonic muscle and rat extensor digitorum longus (EDL) muscle, and estimated the number of pump units by titration with thapsigargin, a high-affinity, specific inhibitor of the SR Ca-ATPase. The Ca2+ uptake rate averaged 70.9 +/- 9.5 mumol min -1 per g tissue for the toad fish sonic muscle, and 73.5 +/- 3.7 mumol min -1 g-1 for rat EDL. The capacity for Ca2+ -oxalate uptake was 161 +/- 20 mumol g -1 and 33 +/- 2 mumol g -1 for toadfish sonic muscle and rat EDL, respectively. Thus, the rates of Ca2+ uptake were similar in the two muscles, but the toadfish sonic muscle had about five times the capacity of the rat EDL. The number of pumps as estimated by thapsigargin titration was 68 +/- 4 nmol of Ca-ATPase per g tissue in the toadfish, and 42 +/- 5 nmol Ca-ATPase per g tissue in the rat EDL. The turnover number, defined as the Ca2+ uptake divided by the number of pumps, was 1065 +/- 150 min -1 for toadfish and 1786 +/- 230 min -1 for rat EDL (p < 0.05) at 37 degrees C. The Ca2+ uptake rate of toadfish sonic muscle at 22 degree C, a typical temperature for calling toadfish, averaged 42 +/- 1% of its rate at 37 degree C. At these operating temperatures, the toadfish SR is likely to be slower than the rat fast-twitch SR, yet the toadfish sonic muscle supports more rapid contractions. One explanation for this is that the voluminous SR provides activator Ca2+ for contraction, but the abundant parvalbumin plays a major role in relaxation.

The effects of seasonal hypertrophy and atrophy on fiber morphology, metabolic substrate concentration and sound characteristics of the weakfish sonic muscle.

Male weakfish Cynoscion regalis possess highly specialized, bilateral, striated sonic muscles used in sound production associated with courtship. Androgen-driven hypertrophy of the muscles during the late spring spawning period results in a tripling of sonic muscle mass followed by post-spawning atrophy. This study examined the morphological and biochemical changes underlying seasonal changes in sonic muscle mass and the functional effects of these on contraction as measured by sound production. Sonic muscle fiber cross-sectional area (CSA) increased significantly during the period of hypertrophy and then decreased by nearly 60%. Both the CSA of the contractile cylinder and that of the peripheral sarcoplasm decreased significantly by late summer, with the peripheral ring of sarcoplasm virtually disappearing. Muscle protein content followed a similar trend, suggesting a major loss of structural elements during atrophy. Muscle glycogen and lipid content decreased precipitously in early June during the period of maximal sound production. Sound pressure level increased and sound pulse duration decreased with increasing sonic muscle mass, indicating that sonic muscle fibers contract with greater force and shorter duration during the spawning season. Neither the pulse repetition rate nor the number of pulses varied seasonally or with muscle mass, suggesting that the effects of steroids on the acoustic variables are more pronounced peripherally than in the central nervous system. Seasonal sonic muscle hypertrophy, therefore, functions as a secondary sexual characteristic that maximizes vocalization amplitude during the spawning period.

Embryonic, juvenile, and adult development of the toadfish sonic muscle.

BACKGROUND: Sonic muscle fibers intrinsic to the swim bladder of the oyster toadfish Opsanus tau proliferate throughout adult life and have an unusual radial morphology: alternating ribbons of sarcoplasmic reticulum (SR) and myofibrils surround a central core of sarcoplasm. Large fibers in adults form multiple cores, fragment, and appear to divide into smaller, more energy efficient units. METHODS: We examined embryonic to adult development of sonic muscle using electron and light microscopy and focused on the incidence of satellite cells (SC). RESULTS: Muscle fibers form late in the larval period from myoblasts, which do not appear to fuse into myotubes, but enlarge and differentiate myofibrils in a single patch. The SR differentiates from the outside inward, separating the myofibrils into bundles of varying thickness, which often exceed the thickness seen in adults. SCs in juveniles and adults have a sparse cytoplasm and a heterochromatic nucleus. The % SC nuclei (SC nuclei/total nuclei) decreases from a high of 88% in larvae to a low of 1% in adults although the adult average is 10%. No embryonic type fibers in the process of differentiating myofibrils were seen in adults. Small immature fibers, which had not yet formed the central core, have a complete radially organized contractile cylinder. CONCLUSIONS: Immature muscle fibers formed embryonically in the larval period have a different morphology from immature fibers in adults, suggesting that splitting rather than SCs is a major source of new fibers in adults.

Lateralization of pectoral stridulation sound production in the channel catfish.

Sounds of the channel catfish Ictalurus punctatus were found to consist of a rapid series of pulses produced by rubbing a ridged process on the first pectoral spine against the rough surface of a groove in the pectoral girdle during fin abduction. Although sounds can be made with either fin, approximately half of the individuals exhibited a fin preference, and 90% of these preferred the right fin. Unlike examples of handedness in other invertebrates and fishes, this preference is not simply a matter of anatomical asymmetry, but as in humans, reflects a preference between two equally developed limbs.

Autoradiographic localization of dihydrotestosterone and testosterone concentrating neurons in the brain of the oyster toadfish.

Vertebrate species with male mating calls or songs tend to have sexually dimorphic sonic neurons that concentrate gonadal steroids. The distribution of [3H]dihydrotestosterone- and testosterone-concentrating neurons was examined in oyster toadfish (Opsanus tau), males of which produce a courtship boatwhistle call. Labeled cells in the forebrain were found in the posterior nucleus of the dorsal telencephalon (Dp), a pallial structure, the supracommissural nucleus of the ventral telencephalon (Vs), nucleus propticus parvocellularis anterior (PPa) and other preoptic nuclei, the ventral, dorsal and caudal hypothalamus. Positive brainstem areas included the optic tectum, torus semicircularis, nucleus lateralis valvula, a periventricular nucleus of the rostral medulla and the inferior reticular formation. Compared to estrogen, androgens labeled fewer sites in the forebrain and more in the brainstem. Two of the positive sites, Vs and PPa, have been implicated in boatwhistle production. Many sites that connect to these areas in teleosts likewise concentrate steroids. Unlike the situation in frogs, birds, and one other teleost, the toadfish sonic motor nucleus did not concentrate androgens. Androgen labeling in the posterior nucleus of the dorsal telencephalon represents the first autoradiographic demonstration of steroid concentration in the pallium of a teleost forebrain.

A Golgi and horseradish peroxidase study of the sonic motor nucleus of the oyster toadfish.

The sonic motor nucleus (SMN) of the oyster toadfish Opsanus tau, a single midline structure in the occipital spinal cord and caudal medulla, contains large electrically-coupled motoneurons. Although interpretation is complicated by multiyear growth in soma size, neurons in males may be either large (L) or small (S), whereas females have exclusively S neurons. Golgi stains have allowed separation of five neuron variants (rostral, dorsal, stellate, ventral and caudal) which differ in location, soma shape and size, and direction and pattern of dendritic branching. All variants are present in L and S males and in females, and retrograde transport of horseradish peroxidase indicates that all variants are motoneurons. The SMN is organized into three horizontal layers with rostral and dorsal neurons forming a rostrocaudally arrayed network across the dorsal-dorsolateral surface. Stellate cells are found in the middle layer, and ventral cells with laterally directed dendrites that exit the SMN line the inferior surface. Caudal neurons with caudally directed exiting dendrites are arranged in parallel rows in the caudal fifth of the SMN. We suggest that variant differences in dendritic orientation relate to different patterns of innervation by multiple afferents to the SMN and function to maximize contacts between neurons as a means of facilitating synchronization within the nucleus. Sexual dimorphism has been demonstrated to a minor degree: all variants have larger somas in L fish than S fish, but no difference has been found in primary dendrite diameter. Larger somas would potentially support the greater amount of sound production by nesting males who produce a mating boatwhistle call. Equivalent dendrite diameter in females, who are just as likely as males to grunt, an agonistic call, suggests that female Opsanus have a well developed sonic circuitry compared to Porichthys, another toadfish in which females are typically silent.

Sound production evoked by electrical stimulation of the forebrain in the oyster toadfish.

In mammals, birds and amphibians the neural pathways controlling sound production descend from higher centers in the forebrain, whereas in fishes only brainstem and spinal centers have been explicitly implicated in sound production. We now report that electrical stimulation of the forebrain of the oyster toadfish (Opsanus tau) readily evokes both the agonistic grunt and the courtship boatwhistle. Boatwhistles are more realistic than ones previously evoked from lower centers. Positive stimulation sites are localized in the preoptic area (nucleus preopticus parvocellularis anterior) and the supra-commissural nucleus of the ventral telencephalon, a likely homologue of the amygdala. Both sites contain gonadal steroid-concentrating neurons and play a central role in fish courtship behavior. Evoked sounds form a continuum from knock grunts, burst grunts, transition boatwhistles to complete boatwhistles; sound pressure level (SPL), fundamental frequency and duration increase consistently within the continuum. For all sound types, SPLs exhibit the smallest variation (coefficients of variation of 2.7 to 5.7%), fundamental frequency is intermediate (5 to 13%) and durations vary most widely (18 to 60%). Boatwhistles, with the smallest variation and greatest amplitude, are likely generated by a maximal output of the CNS and sonic muscles. Grunt SPLs however, vary over a range of 26 dB for all fish and by as much as 18 dB in an individual; suggesting recruitment of variable numbers of motor units despite electrical coupling within the sonic motor nucleus.

Localization of swimbladder and pectoral motoneurons involved in sound production in pimelodid catfish.

We localized the motoneurons and occipital and true spinal innervation of sound-producing organs in pimelodid catfish. Pimelodids have a stridulatory organ composed of the pectoral girdle and the first pectoral fin ray, a swimbladder with extrinsic muscles to produce drumming sounds, and a tensor tripodis (TT) muscle that inserts on the swimbladder. Sonic muscles are innervated by three branches (rostral, dorsal and caudal) of the occipital nerve (Oc) and the first two true spinal nerves (S1 and 2): pectoral spine muscles (abductor, adductor and ventral rotator) by the rostral branch of Oc and S1 and 2, drumming muscle by the caudal branch of Oc and twigs of the S1 and 2, and TT by the dorsal branch of Oc. Sonic nuclei from ipsilateral medial, intermediate and ventrolateral columns in the caudal medulla and spinal cord. Pectoral neurons form a ventrolateral motor column, and neurons for the first spine occupy the rostral part of the column. The medial division of the swimbladder drumming motor nucleus (DMm) is situated on the midline between the central canal and the medial longitudinal fasciculus. The rostral pole of the DM nucleus expands ventrolaterally to include a population of neurons of intermediate position (DMi). The TT nucleus also assumes an intermediate position ventrolateral to DMm. The pectoral, TT, and DMi have a restricted rostrocaudal extent, whereas DMm extends further caudally. These data demonstrate that fish can evolve multiple sonic motor nuclei and that sound producing organs can be innervated in parallel by occipital and spinal nerves.

Localization of pectoral fin motoneurons (sonic and hovering) in the croaking gourami Trichopsis vittatus.

The pectoral fin of the croaking gourami, Trichopsis vittatus, has become modified as a sound-producing organ and retains its original function in locomotion and hovering. We used retrograde transport of horseradish peroxidase to localize sonic motoneurons in Trichopsis. Betta splendens, a related nonsonic gourami with unspecialized pectoral fins, served as a control. A single injection into Trichopsis epaxial muscle labeled a dorsal motor column of large cells (mean of 16.3 microns) ventrolateral to the central canal. Pectoral motoneurons formed a ventrolateral spinal motor column of smaller neurons (means from 7.7 to 11.9 microns, depending upon fish size), of about 2 mm in rostrocaudal extent, starting in the caudal medulla. Our data suggest that motoneurons for different pectoral muscles are segregated into rostrocaudal pools within the column. Distribution, morphology and size of motoneurons were similar between Trichopsis and Betta, and there was no evidence of a distinct population of neurons which might be specialized exclusively for sound production. These data suggest that a fish can evolve a specialized end organ without major reorganization of the central nervous system.