What the Toadfish Ear Tells the Toadfish Brain About Sound.

Of the three, paired otolithic endorgans in the ear of teleost fishes, the saccule is the one most often demonstrated to have a major role in encoding frequencies of biologically relevant sounds. The toadfish saccule also encodes sound level and sound source direction in the phase-locked activity conveyed via auditory afferents to nuclei of the ipsilateral octaval column in the medulla. Although paired auditory receptors are present in teleost fishes, binaural processes were believed to be unimportant due to the speed of sound in water and the acoustic transparency of the tissues in water. In contrast, there are behavioral and anatomical data that support binaural processing in fishes. Studies in the toadfish combined anatomical tract-tracing and physiological recordings from identified sites along the ascending auditory pathway to document response characteristics at each level. Binaural computations in the medulla and midbrain sharpen the directional information provided by the saccule. Furthermore, physiological studies in the central nervous system indicated that encoding frequency, sound level, temporal pattern, and sound source direction are important components of what the toadfish ear tells the toadfish brain about sound.

Does the magnocellular octaval nucleus process auditory information in the toadfish, Opsanus tau?

Previous work on auditory processing in Opsanus tau has focused on the descending octaval nucleus; however, the magnocellular octaval nucleus receives similar inputs from the otolithic endorgans. The purpose of this study was to assess whether cells in any of the three subdivisions of the magnocellular nucleus respond to auditory frequencies and encode sound source direction. Extracellular recording sites were chosen based on anatomical landmarks, and neurobiotin injections confirmed the location of auditory sites in subdivisions of the magnocellular nucleus. In general, the auditory cells in M2 and M3 responded best to frequencies at or below 100 Hz. Most auditory cells responded well to directional stimuli presented along axes in the horizontal plane. Cells in M3 (not M2) also responded to lateral line stimulation, consistent with otolithic endorgan and lateral line inputs to M3. The convergence of auditory and lateral line inputs in M3, the lack of Mauthner cells in this species, and previous evidence that the magnocellular nucleus does not contribute to ascending auditory pathways suggest to us that the large cells of M3 may play a role in rapid behavioral responses to particle motion stimuli in oyster toadfish.

Specialization for underwater hearing by the tympanic middle ear of the turtle, Trachemys scripta elegans.

Turtles, like other amphibious animals, face a trade-off between terrestrial and aquatic hearing. We used laser vibrometry and auditory brainstem responses to measure their sensitivity to vibration stimuli and to airborne versus underwater sound. Turtles are most sensitive to sound underwater, and their sensitivity depends on the large middle ear, which has a compliant tympanic disc attached to the columella. Behind the disc, the middle ear is a large air-filled cavity with a volume of approximately 0.5 ml and a resonance frequency of approximately 500 Hz underwater. Laser vibrometry measurements underwater showed peak vibrations at 500-600 Hz with a maximum of 300 µm s(-1) Pa(-1), approximately 100 times more than the surrounding water. In air, the auditory brainstem response audiogram showed a best sensitivity to sound of 300-500 Hz. Audiograms before and after removing the skin covering reveal that the cartilaginous tympanic disc shows unchanged sensitivity, indicating that the tympanic disc, and not the overlying skin, is the key sound receiver. If air and water thresholds are compared in terms of sound intensity, thresholds in water are approximately 20-30 dB lower than in air. Therefore, this tympanic ear is specialized for underwater hearing, most probably because sound-induced pulsations of the air in the middle ear cavity drive the tympanic disc.

Computerized tomography of the otic capsule and otoliths in the oyster toadfish, Opsanus tau.

The neurocranium of the toadfish (Opsanus tau) exhibits a distinct translucent region in the otic capsule (OC) that may have functional significance for the auditory pathway. This study used ultrahigh resolution computerized tomography (100 µm voxels) to compare the relative density of three sites along the OC (dorsolateral, midlateral, and ventromedial) and two reference sites (dorsal: supraoccipital crest; ventral: parasphenoid bone) in the neurocranium. Higher attenuation occurs where structural density is greater; thus, we compared the X-ray attenuations measured, which provided a measure of relative density. The maximum attenuation value was recorded for each of the five sites (x and y) on consecutive sections throughout the OC and for each of the three calcareous otoliths associated with the sensory maculae (lagena, saccule, and utricle) in the OC. All three otoliths had higher attenuations than any sites in the neurocranium. Both dorsal and ventral reference sites (supraoccipital crest and parasphenoid bone, respectively) had attenuation levels consistent with calcified bone and had relatively small, irregular variations along the length of the OC in all individuals. The lowest relative attenuations (lowest densities) occurred consistently at the three sites along the OC. In addition, the lowest attenuations measured along the OC occurred at the ventromedial site around the saccular otolith for all seven fish. The decrease in bone density along the OC is consistent with the hypothesis that there is a low-density channel in the skull to facilitate transmission of acoustic stimuli to the auditory endorgans of the ear.

Gamma-aminobutyric acid is a neurotransmitter in the auditory pathway of oyster toadfish, Opsanus tau.

Binaural computations involving the convergence of excitatory and inhibitory inputs have been proposed to explain directional sharpening and frequency tuning documented in the brainstem of a teleost fish, the oyster toadfish (Opsanus tau). To assess the presence of inhibitory neurons in the ascending auditory circuit, we used a monoclonal antibody to GABA to evaluate immunoreactivity at three levels of the circuit: the first order descending octaval nucleus (DON), the secondary octaval population (dorsal division), and the midbrain torus semicircularis. We observed a subset of immunoreactive (IR) cells and puncta distributed throughout the neuropil at all three locations. To assess whether contralateral inhibition is present, fluorescent dextran crystals were inserted into dorsal DON to fill contralateral, commissural inputs retrogradely prior to GABA immunohistochemistry. GABA-IR somata and puncta co-occurred with retrogradely filled, GABA-negative auditory projection cells. GABA-IR projection cells were more common in the dorsolateral DON than in the dorsomedial DON, but GABA-IR puncta were common in both dorsolateral and dorsomedial divisions. Our findings demonstrate that GABA is present in the ascending auditory circuit in the brainstem of the toadfish, indicating that GABA-mediated inhibition participates in shaping auditory response characteristics in a teleost fish as in other vertebrates.

Physiological evidence for binaural directional computations in the brainstem of the oyster toadfish, Opsanus tau (L.).

Comparisons of left and right auditory input are required for sound source localization in most terrestrial vertebrates. Previous physiological and neuroanatomical studies have indicated that binaural convergence is present in the ascending auditory system of the toadfish. In this study, we introduce a new technique, otolith tipping, to reversibly alter directional auditory input to the central nervous system of a fish. The normal directional response pattern (DRP) was recorded extracellularly for auditory cells in the first-order descending octaval nucleus (DON) or the midbrain torus semicircularis (TS) using particle motion stimuli in the horizontal and mid-sagittal planes. The same stimuli were used during tipping of the saccular otolith to evaluate changes in the DRPs. Post-tipping DRPs were generated and compared with the pre-tipping DRPs to ensure that the data had been collected consistently from the same unit. In the DON, ipsilateral or contralateral tipping most often eliminated spike activity, but changes in spike rate (+/-) and DRP shape were also documented. In the TS, tipping most often caused a change in spike rate (+/-) and altered the shape or best axis of the DRP. The data indicate that there are complex interactions of excitatory and inhibitory inputs in the DON and TS resulting from the convergence of binaural inputs. As in anurans, but unlike other terrestrial vertebrates, binaural processing associated with encoding the direction of a sound source begins in the first-order auditory nucleus of this teleost.

Directional and frequency response characteristics in the descending octaval nucleus of the toadfish (Opsanus tau).

This study is a continuation of a long-term investigation of the auditory circuit in the oyster toadfish, Opsanus tau. Input from the auditory periphery projects to the ipsilateral descending octaval nucleus (DON). Ipsilateral and contralateral DONs project to the auditory midbrain, where a previous study indicated that both frequency tuning and directional sharpening are present. To better understand the transformation of auditory information along the auditory pathway, we have examined over 400 units in the DON to characterize frequency and directional information encoded in the dorsolateral division of the nucleus. Background activity was primarily low (<10 spikes/s) or absent. The maximum coefficient of synchronization was equivalent to the periphery (R = 0.9) and substantially better than in the midbrain. The majority of DON units (79%) responded best to stimulus frequencies of 84-141 Hz and were broadly tuned. DON cells retain or enhance the directional character of their peripheral input (s); however, characteristic axes were distributed in all quadrants around the fish, providing further evidence that binaural computations may first occur in the DON of this species.

Sharpening of directional responses along the auditory pathway of the oyster toadfish, Opsanus tau.

Our previous studies have shown that the peripheral auditory system of the toadfish encodes the direction of a sound source. Here, we compare directional responses of peripheral saccular afferents, cells in the descending octaval nucleus (DON) of the medulla, and the torus semicircularis (TS) of the midbrain. Recording locations in the brain were labeled with neurobiotin to confirm the site. To compare directional responses among cells, we calculated an index [sharpening ratio (SR)] that weights the relative strength of responses to the best direction for that cell and to the adjacent stimulus angles tested. Unsharpened saccular afferents tend to have a cosinusoidal directional response pattern (DRP) with an expected SR of 0.87. In DON, more than 60% of the cells exhibited directional sharpening (defined as SR <0.8). In TS, more than 80% of the cells exhibited directional sharpening. We conclude that directional auditory sharpening first occurs in DON and some additional sharpening occurs in the ascending pathway to the midbrain, particularly in azimuth. The sharpening of directional selectivity is likely to be an important component of the neural computations underlying directional hearing.

Projections to bimodal sites in the torus semicircularis of the toadfish, Opsanus tau.

Bimodal cells in the torus semicircularis of the toadfish respond to both directional acoustic stimuli and hydrodynamic stimuli. Our previous physiological work indicated that bimodal cells may be distributed throughout the torus semicircularis. In this study, neurobiotin was used to compare the distribution of auditory-only and bimodal sites and to assess the inputs to those sites. A brief neurobiotin injection with short survival time was used to document the recording location. In other fish, a longer injection and survival time was used at an auditory-only or a bimodal site to fill the axons of the medullary inputs. Auditory-only sites were located in the most dorsal and medial sites in nucleus centralis. Bimodal sites were identified within both nucleus centralis and nucleus ventrolateralis. The greatest number of retrogradely filled cell bodies was found in the descending octaval nucleus following injection at auditory-only recording sites in nucleus centralis. In contrast, retrogradely filled cell bodies were found in both the descending octaval nucleus and the lateral line nucleus medialis following injection at bimodal sites in nucleus centralis or nucleus ventrolateralis. Retrogradely filled cell bodies were located in the dorsal and ventral divisions of the secondary octaval population from injections at either bimodal or auditory-only sites. The secondary octaval population has been implicated in auditory processing based on previous studies of both auditory specialist and auditory generalist fishes; however, this study is the first to reveal the potential role of the secondary octaval population in directional hearing in a fish. Relatively large numbers of retrogradely filled cells around the lateral lemniscus at consistent locations in the medulla indicate that a perilemniscal cell group also might be a component of the directional hearing circuit.