Vocal production complexity correlates with neural instructions in the oyster toadfish (Opsanus tau).

Sound communication is fundamental to many social interactions and essential to courtship and agonistic behaviours in many vertebrates. The swimbladder and associated muscles in batrachoidid fishes (midshipman and toadfish) is a unique vertebrate sound production system, wherein fundamental frequencies are determined directly by the firing rate of a vocal-acoustic neural network that drives the contraction frequency of superfast swimbladder muscles. The oyster toadfish boatwhistle call starts with an irregular sound waveform that could be an emergent property of the peripheral nonlinear sound-producing system or reflect complex encoding in the central nervous system. Here, we demonstrate that the start of the boatwhistle is indicative of a chaotic strange attractor, and tested whether its origin lies in the peripheral sound-producing system or in the vocal motor network. We recorded sound and swimbladder muscle activity in awake, freely behaving toadfish during motor nerve stimulation, and recorded sound, motor nerve and muscle activity during spontaneous grunts. The results show that rhythmic motor volleys do not cause complex sound signals. However, arrhythmic recruitment of swimbladder muscle during spontaneous grunts correlates with complex sounds. This supports the hypothesis that the irregular start of the boatwhistle is encoded in the vocal pre-motor neural network, and not caused by peripheral interactions with the sound-producing system. We suggest that sound production system demands across vocal tetrapods have selected for muscles and motorneurons adapted for speed, which can execute complex neural instructions into equivalently complex vocalisations.

The structure of the caudal wall of the zebrafish (Danio rerio) swim bladder: evidence of localized lamellar body secretion and a proximate neural plexus.

In this study, we present a morphological description of the fine structure of the tissues composing the caudal tip of the adult zebrafish swim bladder and an immunochemical survey of the innervation at this site. The internal aspect of the caudal tip is lined by an epithelium specialized to secrete surfactant into the lumen as evinced by the exocytosis of lamellar bodies. The sole innervation to this region consists of a neural plexus, present on the external surface, of nitric oxide synthase-positive (nNOS) neuronal cell bodies that are contacted by axon terminals, some containing neuropeptide Y and vasoactive intestinal polypeptide. As the specialized epithelium and neural plexus are coincident and of common extent, we suggest that the morphological relationship between the two elements allows the nervous system to affect surfactant processing, possibly through a paracrine mechanism.

Fast drum strokes: novel and convergent features of sonic muscle ultrastructure, innervation, and motor neuron organization in the Pyramid Butterflyfish (Hemitaurichthys polylepis).

Sound production that is mediated by intrinsic or extrinsic swim bladder musculature has evolved multiple times in teleost fishes. Sonic muscles must contract rapidly and synchronously to compress the gas-filled bladder with sufficient velocity to produce sound. Muscle modifications that may promote rapid contraction include small fiber diameter, elaborate sarcoplasmic reticulum (SR), triads at the A-I boundary, and cores of sarcoplasm. The diversity of innervation patterns indicate that sonic muscles have independently evolved from different trunk muscle precursors. The analysis of sonic motor pathways in distantly related fishes is required to determine the relationships between sonic muscle evolution and function in acoustic signaling. We examined the ultrastructure of sonic and adjacent hypaxial muscle fibers and the distribution of sonic motor neurons in the coral reef Pyramid Butterflyfish (Chaetodontidae: Hemitaurichthys polylepis) that produces sound by contraction of extrinsic sonic muscles near the anterior swim bladder. Relative to adjacent hypaxial fibers, sonic muscle fibers were sparsely arranged among the endomysium, smaller in cross-section, had longer sarcomeres, a more elaborate SR, wider t-tubules, and more radially arranged myofibrils. Both sonic and non-sonic muscle fibers possessed triads at the Z-line, lacked sarcoplasmic cores, and had mitochondria among the myofibrils and concentrated within the peripheral sarcoplasm. Sonic muscles of this derived eutelost possess features convergent with other distant vocal taxa (other euteleosts and non-euteleosts): small fiber diameter, a well-developed SR, and radial myofibrils. In contrast with some sonic fishes, however, Pyramid Butterflyfish sonic muscles lack sarcoplasmic cores and A-I triads. Retrograde nerve label experiments show that sonic muscle is innervated by central and ventrolateral motor neurons associated with spinal nerves 1-3. This restricted distribution of sonic motor neurons in the spinal cord differs from many euteleosts and likely reflects the embryological origin of sonic muscles from hypaxial trunk precursors rather than occipital somites.

Morphology and innervation of the teleost physostome swim bladders and their functional evolution in non-teleostean lineages.

Swim bladders and lungs are homologous structures. Phylogenetically ancient actinopterygian fish such as Cladistians (Polypteriformes), Ginglymods (Lepisosteids) and lungfish have primitive lungs that have evolved in the Paleozoic freshwater earliest gnathostomes as an adaptation to hypoxic stress. Here we investigated the structure and the role of autonomic nerves in the physostome swim bladder of the cyprinid goldfish (Carassius auratus) and the respiratory bladder of lepisosteids: the longnose gar and the spotted gar (Lepisosteus osseus and L. oculatus) to demonstrate that these organs have different innervation patterns that are responsible for controlling different functional aspects. The goldfish swim bladder is a richly innervated organ mainly controlled by cholinergic and adrenergic innervation also involving the presence of non-adrenergic non-cholinergic (NANC) neurotransmitters (nNOS, VIP, 5-HT and SP), suggesting a simple model for the regulation of the swim bladder system. The pattern of the autonomic innervation of the trabecular muscle of the Lepisosteus respiratory bladder is basically similar to that of the tetrapod lung with overlapping of both muscle architecture and control nerve patterns. These autonomic control elements do not exist in the bladders of the two species studied since they have very different physiological roles. The ontogenetic origin of the pulmonoid swim bladder (PSB) of garfishes may help understand how the expression of these autonomic control substances in the trabecular muscle is regulated including their interaction with the corpuscular cells in the respiratory epithelium of this bimodal air-breathing fish.

Autonomic control of the swimbladder.

The swimbladder of teleost fishes is the primary organ for controlling whole-body density, and thus buoyancy. The volume of gas in the swimbladder is adjusted to bring the organism to near neutral buoyancy at a particular depth. Swimbladder morphology varies widely among teleosts, but all species are capable of inflating and deflating this organ under reflex control by the autonomic nervous system, to achieve neutral buoyancy. Here we review the control of effectors within the swimbladder, including acid-secreting cells, vasculature and musculature, that are involved in determining gas volume. This control system is complex. It incorporates the "classical" efferent elements of the autonomic nervous system, the spinal autonomic and cranial autonomic limbs and their neurotransmitters (typically noradrenaline (NA)/adrenaline (ADR), and acetylcholine, respectively), but also non-adrenergic, non-cholinergic neurotransmitters such as peptides, purines and nitric oxide. The detailed patterns of autonomic innervation of swimbladder effectors are not well understood, nor are the relationships of terminals releasing non-adrenergic, non-cholinergic neurotransmitters onto these effectors. Furthermore, in most cases the complement of postjunctional receptor subtypes activated by adrenergic, cholinergic and other neurotransmitters, and the biological effects of these neurochemicals, have not been completely established. In order to clarify some of these issues and to provide insight into basic principles underlying autonomic control of swimbladder function, we propose the zebrafish as a potentially useful model teleost.

Adrenergic control of swimbladder deflation in the zebrafish (Danio rerio).

Many teleosts actively regulate buoyancy by adjusting gas volume in the swimbladder. In physostomous fishes such as the zebrafish, a connection is maintained between the swimbladder and the oesophagus via the pneumatic duct for the inflation and deflation of this organ. Here we investigated the role of adrenergic stimulation of swimbladder wall musculature in deflation of the swimbladder. Noradrenaline (NA), the sympathetic neurotransmitter (dosage 10(-6) to 10(-5) mol l(-1)), doubled the force of smooth muscle contraction in isolated tissue rings from the anterior chamber, caused a doubling of pressure in this chamber in situ, and evoked gas expulsion through the pneumatic duct, deflating the swimbladder to approximately 85% of the pre-NA volume. These effects were mediated by beta-adrenergic receptors, representing a novel role for these receptors in vertebrates. No effects of adrenergic stimulation were detected in the posterior chamber. In a detailed examination of the musculature and innervation of the swimbladder to determine the anatomical substrate for these functional results, we found that the anterior chamber contained an extensive ventral band of smooth muscle with fibres organized into putative motor units, richly innervated by tyrosine hydroxylase-positive axons. Additionally, a novel arrangement of folds in the lumenal connective tissue in the wall of the anterior chamber was described that may permit small changes in muscle length to cause large changes in effective wall distensibility and hence chamber volume. Taken together, these data strongly suggest that deflation of the zebrafish swimbladder occurs primarily by beta-adrenergically mediated contraction of smooth muscle in the anterior chamber and is under the control of the sympathetic limb of the autonomic nervous system.

Nervous control of fish swimbladders.

The swimbladder of teleost fish receives a rich and complex innervation by nerve fibres of the autonomic nervous system. While an understanding of the form and function of a non-adrenergic, non-cholinergic innervation is slowly emerging, the pattern of control by the "classical" cholinergic and adrenergic innervation is becoming relatively well understood. This short review describes the autonomic innervation patterns, and attempts to summarise the role of cholinergic and adrenergic pathways in the control of gas secretion and resorption in the teleost swimbladder.

Development of the swimbladder and its innervation in the zebrafish, Danio rerio.

Many teleosts including zebrafish, Danio rerio, actively regulate buoyancy with a gas-filled swimbladder, the volume of which is controlled by autonomic reflexes acting on vascular, muscular, and secretory effectors. In this study, we investigated the morphological development of the zebrafish swimbladder together with its effectors and innervation. The swimbladder first formed as a single chamber, which inflated at 1-3 days posthatching (dph), 3.5-4 mm body length. Lateral nerves were already present as demonstrated by the antibody zn-12, and blood vessels had formed in parallel on the cranial aspect to supply blood to anastomotic capillary loops as demonstrated by Tie-2 antibody staining. Neuropeptide Y-(NPY-) like immunoreactive (LIR) fibers appeared early in the single-chambered stage, and vasoactive intestinal polypeptide (VIP)-LIR fibers and cell bodies developed by 10 dph (5 mm). By 18 dph (6 mm), the anterior chamber formed by evagination from the cranial end of the original chamber; both chambers then enlarged with the ductus communicans forming a constriction between them. The parallel blood vessels developed into an arteriovenous rete on the cranial aspect of the posterior chamber and this region was innervated by zn-12-reactive fibers. Tyrosine hydroxylase- (TH-), NPY-, and VIP-LIR fibers also innervated this area and the lateral posterior chamber. Innervation of the early anterior chamber was also demonstrated by VIP-LIR fibers. By 25-30 dph (8-9 mm), a band of smooth muscle formed in the lateral wall of the posterior chamber. Although gas in the swimbladder increased buoyancy of young larvae just after first inflation, our results suggest that active control of the swimbladder may not occur until after the formation of the two chambers and subsequent development and maturation of vasculature, musculature and innervation of these structures at about 28-30 dph.

Structure and autonomic innervation of the swim bladder in the zebrafish (Danio rerio).

Many teleosts actively regulate buoyancy by using a gas-filled swim bladder, which is thought to be under autonomic control. Here we investigated the swim bladder in the zebrafish to determine possible mechanisms of gas-content regulation. Fluorescently labelled phalloidin revealed myocytes that appeared to form a possible sphincter at the junction of the pneumatic duct and esophagus. Myocytes also formed thick bands along the ventral surface of the anterior chamber and bilaterally along the posterior chamber. Thinner layers of myocytes were located elsewhere. Staining of peroxidase within erythrocytes revealed a putative rete and smaller blood vessels in muscle bands and elsewhere. The antibodies zn-12, a general neuronal marker, and SV2, a synaptic vesicle marker labelling presynaptic terminals, revealed widespread innervation of the swim bladder system. Widespread innervation of the swim bladder was also indicated by acetylcholinesterase histochemistry, but choline acetyltransferase-immunoreactive (-IR) somata and fibers were limited to the junction of the pneumatic duct and esophagus. In contrast, varicose tyrosine hydroxylase-IR fibers innervated muscles and blood vessels throughout the system. Neuropeptide Y-IR somata were located near the junction of the duct and esophagus and varicose fibers innervated muscles and vasculature of the posterior chamber and duct. Vasoactive intestinal polypeptide immunoreactivity was abundant throughout the anterior chamber but sparsely distributed elsewhere. Serotonin-IR fibers and varicosities were located only along blood vessels near the junction of the pneumatic duct and posterior chamber. Our results suggest that the zebrafish swim bladder is a complex and richly innervated organ and that buoyancy-regulating effectors may be controlled by multiple populations of autonomic neurons.

Spinal nerve innervation to the sonic muscle and sonic motor nucleus in red piranha, Pygocentrus nattereri (Characiformes, Ostariophysi).

The red piranha, Pygocentrus nattereri, produces sounds by rapid contractions of a pair of extrinsic sonic muscles. The detailed innervation pattern of the sonic muscle of the red piranha was investigated. The sonic muscle is innervated by branches (sonic branches) of the third (S3so), fourth (S4so), and fifth (S5so) spinal nerves. The average total number of nerve fibers contained in the right sonic branches (n = 5; standard length, SL, 71-85 mm) was 151.8 (standard deviation, SD, 28.3). The occipital nerve did not innervate the sonic muscle. The sonic motor nucleus (SMN) in the piranha was identified by tracer methods using wheat germ agglutinin-conjugated horseradish peroxidase; labeled sonic motor neurons were only observed on the side ipsilateral to the sonic muscle injected with the tracer. In the transverse sections, the labeled sonic motor neurons were located in the dorsal zone (mainly large and medium neurons) and in the ventral zone (mainly small neurons) of the ventral horn. In the horizontal sections, the labeled neurons formed a rostrocaudally elongated SMN from the level of the caudal part of the second spinal nerve root to the intermediate region between the fifth and sixth spinal nerve roots. The average number of the labeled neurons (n = 5; SL, 64-87 mm) was 152.6 (SD, 7.3). We conclude that the sonic muscles of the piranha are innervated by approximately 300 sonic motor neurons located only in the spinal cord.

Sonic motor pathways in piranhas with a reassessment of phylogenetic patterns of sonic mechanisms among teleosts.

Sound production has evolved independently a number of times among teleost fishes. In most cases, sound is generated by fast contracting muscles that vibrate the swim bladder by way of their direct attachment (intrinsic muscles) or indirectly by way of other skeletal elements (extrinsic muscles). This study focuses on the red and black piranha, Pygocentrus nattereri and Serrasalmus rhombeus (superorder Ostariophysi, Order Characiformes), that have extrinsic swim bladder sonic muscles innervated by the third and fourth spinal nerves. This innervation pattern diverges from that found in most teleosts, including the closely related catfishes (Ostariophysi, Siluriformes), where sonic muscles are innervated by ventral occipital nerve roots that arise just caudal to the vagus nerve. Here, we tested the hypothesis that piranhas would also differ from most other teleosts in the location of their sonic motor neurons. Following biotin labeling of branches of the third and fourth spinal nerves that innervate the sonic muscles in the red and black piranha, sonic motor neurons were identified amongst other non-sonic motor neurons in the central part of the spinal cord, slightly ventrolateral to the central canal. To our knowledge, this is the first example of sonic motor neurons positioned entirely within the spinal cord. In the other species so far studied, sonic motor neurons form well-defined nuclei that extend from far caudal levels of the medulla into the rostral spinal cord and are located either within the ventral motor column or near the midline, close to or just ventrolateral to the fourth ventricle and central canal. A piranha-like pattern may be more widespread among characiforms and is likely present in other teleost orders, e.g., Sciaenidae (drumfishes), that also have sonic muscles innervated by spinal nerves.

Vagal innervation of the air sacs in a songbird, Taenopygia guttata.

The air sacs of birds are thin-walled chambers connected to the lung that act as bellows in the ventilatory mechanism. Physiological evidence exists to suggest that they may contain receptors that are innervated by the vagus nerve, but no morphological study has examined the vagal innervation of these putative structures. To do this, we injected the cervical vagus nerve with choleragenoid and examined the innervation of the air sacs using light and confocal microscopy. We identified vagally innervated structures in the air sac wall that resemble the neuroepithelial bodies (NEBs) described in the airways of many vertebrates. Although NEBs have been proposed to have a dual chemoreceptive and mechanoreceptive role, their specific function in the air sacs of birds remains unclear.

NANC nerves in the respiratory air sac and branchial vasculature of the Indian catfish, Heteropneustes fossilis.

Gill and air sac of the Indian catfish Heteropneustes fossilis harbour a nerve network comprising an innervated system of neuroepithelial endocrine cells; the latter cells are found especially in the gill. A series of antibodies was used for the immunohistochemical detection of neurotransmitters of the neural non-adrenergic, non-cholinergic (NANC) systems such as the sensory neuropeptides (enkephalins), the inhibitory neuropeptide VIP and neuronal nitric oxide synthase (nNOS) responsible for nitric oxide (NO) production which is an inhibitory NANC neurotransmitter. NADPH-diaphorase (NADPH-d) histochemistry was used as marker of nNOS although it is not a specific indicator of constitutively-expressed NOS in gill and air sac tissues. A tyrosine hydroxylase antibody was used to investigate adrenergic innervation. Nitrergic and VIP-positive sensory innervation was found to be shared by gill and air sac. Immunohistochemistry revealed the presence of enkephalins, VIP, NOS and NADPH-d in nerves associated with branchial and air sac vasculature, and in the neuroendocrine cell systems of the gill. Adrenergic nerve fibers were found in some parts of the air sac vasculature. The origin of the nerve fibers remains unclear despite previous findings showing the presence of both NADPH-d and nNOS in the sensory system of the glossopharyngeal and vagus nerves including the branchial structure. Scarce faintly stained nNOS-positive neurons were located in the gill but were never detected in the air sac. These findings lead to the conclusion that a postganglionic innervation of the airways is absent. Mucous goblet cells in the gill were found to express nNOS and those located in the non-respiratory interlamellar areas of the air sac were densely innervated by nNOS-positive and VIP-positive nerve fibers. Our immunohistochemical studies demonstrate that most arteries of the gill and air sac share a NANC (basically nitrergic) innervation which strongly suggests that they are homologous structures.

Sonic motor nucleus and its connections with octaval and lateral line nuclei of the medulla in a rockfish, Sebastiscus marmoratus.

The sonic motor nucleus and its fiber connections were examined in a rockfish, Sebastiscus marmoratus by means of tracer methods using horseradish peroxidase (HRP), biocytin, and carbocyanine dye (DiI). Sebastiscus has a swimbladder and a pair of extrinsic sonic/drumming muscles. The sonic muscle is ipsilaterally innervated by the occipital nerve which is composed of two ventral roots arising from the sonic motor nucleus. The sonic motor neurons are distributed in the most ventral part of the ventral column from the caudal medulla to the rostral spinal cord, and form a ventrally located columnar nucleus. Each neuron in this nucleus possesses a long thick dendrite and several short dendrites. The long dendrite extends dorsolaterally and branches in the lateral funiculus, whereas the short dendrites branch around their cell bodies. After biocytin injections into the sonic motor nucleus, two groups of premotor neurons were retrogradely labeled bilaterally, one in the dorsomedial portion of the descending octaval nucleus (DO) and the other in the medial zone of the reticular formation (RF) in the medulla. The DO premotor neurons were multipolar with several dendrites branching near the cell bodies, and the RF premotor neurons were bipolar. One of the two dendrites of the RF premotor neurons extends laterally into the ventral portion of the DO, and the other dendrite extends into the ventromedial area in the medulla. In the ventromedial dendritic field of the RF premotor neurons, descending fibers arising from the optic tectum (TO) and torus semicircularis (TS) traverse in the tractus tectobulbaris and terminate bilaterally. After DiI insertion into the ventromedial dendritic field, retrogradely labeled neurons were found bilaterally in the TS and TO. The majority of tectal neurons were located in the stratum griseum centrale. These neurons had two short basal dendrites branching in the cell layer and a long apical dendrite extending to the stratum fibrosum et griseum superficiale and stratum opticum. The toral neurons were bipolar and were distributed throughout the TS. Furthermore, biocytin injections into the medial nucleus of the lateral line system revealed that the nucleus projects bilaterally to the RF premotor neurons. These results show that premotor neurons for the sonic motor nucleus are located in the dorsomedial portion of the DO and the medial zone of the RF in the medulla. It is suggested that the sonic motor nucleus receives auditory input via the DO premotor neurons and input from RF premotor neurons which receive lateral line input via the medial nucleus, vestibular input through the lateral dendrite extending into the ventral portion of the DO, and information from the TO and TS via the tractus tectobulbaris.

Sonic/vocal motor pathways in catfishes: comparisons with other teleosts.

Among teleost fishes, representatives of several distantly related groups have sound-producing (sonic/vocal) muscles associated with the swimbladder or pectoral girdle/fin. Here, the diversity of vocal organs and central motor pathways in four families of catfish, order Siluriformes, is compared to that in families from two distantly related orders, the Scorpaeniformes and Batrachoidiformes. Several catfish families have two sonic mechanisms--a swimbladder vibration established by 'drumming muscles' that differ in origin and insertion between families, and a pectoral spine stridulatory apparatus. In ariids, mochokids and doradids, sonic swimbladder muscles originate at various cranial or postcranial elements and insert onto an 'elastic spring' that vibrates the swimbladder, while in pimelodids the muscles insert ventrally at the swimbladder. Sonic motoneurons are located along the midline, ventral to the fourth ventricle/central canal in doradids and mochokids but lateral to the medial longitudinal fasciculus in ariids; pimelodids have motoneurons in both locations. The axonal trajectory for the lateral motoneurons in pimelodids and ariids implies that they are a migrated, midline population of sonic motoneurons. Pectoral spine-associated motoneurons are located in the ventral motor column. Unlike catfishes, a diversity of sonic mechanisms in Scorpaeniformes is not associated with different positions for sonic motoneurons. Cottids (sculpin) lack a swimbladder but have sonic muscles that originate at the occipital cranium and insert at the pectoral girdle; sonic motoneurons are located within the ventral motor column. Some triglids have intrinsic swimbladder muscles, although ontogenetic data indicate a transient association with the pectoral girdle; sonic motoneurons are in the same location as in cottids. Among Batrachoidiformes, all known representatives have intrinsic swimbladder muscles that are never associated with the pectoral girdle and are innervated by midline sonic motoneurons. The results suggest two patterns of organization for sound-producing systems in teleost fishes: pectoral fin/girdle-associated muscles are innervated by sonic motoneurons positioned within the ventral motor column, adjacent to the ventral fasciculus; non-pectoral associated muscles are innervated by sonic motoneurons located on or close to the midline, adjacent to the medial longitudinal fasciculus.

Neuronal encoding of sound direction in the auditory midbrain of the rainbow trout.

Acoustical stimulation causes displacement of the sensory hair cells relative to the otoliths of the fish inner ear. The swimbladder, transforming the acoustical pressure component into displacement, also contributes to the displacement of the hair cells. Together, this (generally) yields elliptical displacement orbits. Alternative mechanisms of fish directional hearing are proposed by the phase model, which requires a temporal neuronal code, and by the orbit model, which requires a spike density code. We investigated whether the directional selective response of auditory neurons in the midbrain torus semicircularis (TS; homologous to the inferior colliculus) is based on spike density and/or temporal encoding. Rainbow trout were mounted on top of a vibrating table that was driven in the horizontal plane to simulate sound source direction. Rectilinear and elliptical (or circular) motion was applied at 172 Hz. Generally, responses to rectilinear and elliptical/circular stimuli (irrespective of direction of revolution) were the same. The response of auditory neurons was either directionally selective (DS units, n = 85) or not (non-DS units, n = 106). The average spontaneous discharge rate of DS units was less than that of non-DS units. Most DS units (70%) had spontaneous activities < 1 spike per second. Response latencies (mode at 18 ms) were similar for both types of units. The response of DS units is transient (19%), sustained (34%), or mixed (47%). The response of 75% of the DS units synchronized to stimulus frequency, whereas just 23% of the non-DS responses did. Synchronized responses were measured at stimulus amplitudes as low as 0.5 nm (at 172 Hz), which is much lower than for auditory neurons in the medulla of the trout, suggesting strong convergence of VIIIth nerve input. The instant of firing of 42% of the units was independent of stimulus direction (shift <15 degrees), but for the other units, a direction dependent phase shift was observed. In the medial TS spatial tuning of DS units is in the rostrocaudal direction, whereas in the lateral TS all preferred directions are present. On average, medial DS units have a broader directional selectivity range, are less often synchronized, and show a smaller shift of the instant of firing as a function of stimulus direction than lateral DS units. DS response characteristics are discussed in relation to different hypotheses. We conclude that the results are more in favor of the phase model.

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.

The primary octaval nuclei and inner ear afferent projections in the otophysan Ictalurus punctatus.

Physical coupling between the swimbladder and the inner ear is believed to contribute to the enhanced hearing abilities of otophysans relative to those of teleosts lacking comparable otic specializations. We hypothesized that the auditory circuitry of otophysans might also exhibit derived features [McCormick and Braford, 1988]. As a first test of this hypothesis, we examined the normal anatomy and inner ear inputs of the primary octaval nuclei in the catfish Ictalurus punctatus. From Nissl-stained sections we conclude that Ictalurus, like other teleosts, has five first-order octaval nuclei: the anterior, magnocellular, tangential, descending, and posterior octaval nuclei. The overall projection pattern of the seven inner ear endorgans, determined using the exsanguinated horseradish peroxidase procedure [McCormick and Braford, 1984], is very similar to that hypothesized as primitive for gnathostome fishes. The otolithic endorgans project to dorsal areas of the descending and anterior nuclei, whereas the macula neglecta and cristae of the semicircular canals project more ventrally. Three zones are recognized within the descending nucleus-the dorsomedial, intermediate, and ventral zones. The dorsomedial zone, which is the main terminus of saccular fibers, has specialized morphological features: it extends far dorsally and contacts the medial portion of the cerebellar crest, and it is partitioned into lateral and medial portions by entering facial nerve fibers. The caudal anterior nucleus is likewise partitioned into lateral and medial portions by the facial nerve. Hypotheses addressing these derived features are presented.

Peripheral and central aspects of the acoustic and lateral line system of a bottom dwelling catfish, Ancistrus sp.

The topographical relationship between the swim bladder, the inner ear, and the otic lateral line was studied in the bottom dwelling catfish, Ancistrus sp. In addition, afferent and efferent subcomponents of the eighth and lateral line nerves were labelled with horseradish peroxidase (HRP) or with differently fluorescing dextran amines. The swim bladder of Ancistrus consists of two separate, transversely oriented parts of each of which is connected to the sinus impar of the inner ears via two Weberian ossicles and the perilymphatic sac. The osseous capsula of the ear has two foramina other than the nerve foramina. One is for the sinus impar. The other foramen, which also separates two fluid-filled spaces, exits where the horizontal canal of the ear contacts the otic lateral line. Both the otic and the postotic lateral line canal run deep below the epidermis. Each canal contains a neuromast that is innervated by the middle lateral line nerve. Further caudally, the otic lateral line canal gives rise to the postotic and finally to the truck canal whose nonossified anterior part travels through an ossified chamber that surrounds the swim bladder. Thus the anterior part of each trunk lateral line canal is in contact with a bipartite sound pressure receiver, the swim bladder. Anterior and posterior lateral line afferents terminate ipsilaterally throughout the neuropil of the electroreceptive lateral line nucleus and the mechanoreceptive nuclei medialis and caudalis of the medulla. Middle lateral line afferents terminate between the projection sites of anterior and posterior lateral line afferents. Some primary mechanosensory anterior lateral line nerve fibers continue into the ipsilateral eminentia granularis and the valvula cerebelli. In the electroreceptive lateral line projection, anterior lateral line fibers terminate more medially and posterior fibers more laterally. This somatotopy is not as clear-cut in the mechanosensory lateral line. Afferents of the sacculus and the lagena terminate predominantly in the saccular nucleus. Afferents of the utriculus, the horizontal canal, and the anterior vertical canal terminate in the magnocellular vestibular nucleus and in the medial octavolateral nucleus. The projection sites of the anterior part and the posterior part of the eighth nerve show little overlap. Eighth nerve projections to the valvula cerebelli are less prominent than the projections from the lateral line. Eighth nerve and lateral line nerve efferents arise from a common nucleus, the octavolateralis efferent nucleus. Axons of efferent cells may divide to supply two or more branches of the eighth nerve and some axons supply both lateral line and eighth nerve endorgans.(ABSTRACT TRUNCATED AT 400 WORDS)

Inter- and intrasexual dimorphisms in the vocal control system of a teleost fish: motor axon number and size.

In one species of vocalizing fish, the plainfin midshipman (Porichthys notatus), large, nest-guarding males ('type I') use striated muscles to produce acoustic communication signals that include short duration (less than 1 s) 'burps' important in agonistic encounters and long duration (in the order of minutes) 'hums' which function in attracting females to nest sites during the breeding season. Females, and a second group of smaller reproductively active males ('type II') that 'sneak' spawn, do not generate hums, although they produce burps. Differences in vocal behavior are paralled by a relative increase of 6-fold in the sonic muscle of type I males. Inter- and intrasexual dimorphisms in sonic muscle mass were matched by those in the cross-sectional area of sonic motor axons, but not by those in number of axons. Thus, axon size was 2- to 3-fold larger in type I males than in females, type II males, or juveniles, none of which differed significantly from each other. Axon number was similar between type I males and females of a similar body size, despite the extreme dimorphism in sonic muscle mass. Axon number, however, was slightly greater (0.1-fold) in type I males and females compared to the smaller-sized juveniles and type II males. Type II males, in comparison to the non-reproductive juvenile males, have gonads that are about 20-fold larger and produce mature sperm. Nevertheless, the two groups resemble each other in body and swimbladder size, as well as sonic motor axon size and number. This suggests that type II males represent a subset of juvenile males that undergo precocious gonadal hypertrophy and spermiogenesis, but retain juvenile-like nongonadal traits. The results are discussed within the context of the development of vertebrate motor systems as well as the evolution of alternative reproductive tactics among teleost fishes.