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In seasonally breeding vertebrates, hormones coordinate changes in nervous system structure and function to facilitate reproductive readiness and success. Steroid hormones often exert their effects indirectly via regulation of neuromodulators, which in turn can coordinate the modulation of sensory input with appropriate motor output. Female plainfin midshipman fish (Porichthys notatus) undergo increased peripheral auditory sensitivity in time for the summer breeding season, improving their ability to detect mates, which is regulated by steroid hormones. Reproductive females also show differences in catecholaminergic innervation of auditory circuitry compared with winter, non-reproductive females as measured by tyrosine hydroxylase (TH), the rate-limiting enzyme in catecholaminergic synthesis. Importantly, catecholaminergic input to the inner ear from a dopaminergic-specific forebrain nucleus is decreased in the summer and dopamine inhibits the sensitivity of the inner ear, suggesting that gonadal steroids may alter auditory sensitivity by regulating dopamine innervation. In this study, we gonadectomized non-reproductive females, implanted them with estradiol (E2) or testosterone (T), and measured TH immunoreactive (TH-ir) fibers in auditory nuclei where catecholaminergic innervation was previously shown to be seasonally plastic. We found that treatment with T, but not E2, reduced TH-ir innervation in the auditory hindbrain. T-treatment also reduced TH-ir fibers in the forebrain dopaminergic cell group that projects to the inner ear, and likely to the auditory hindbrain. Higher T plasma in the treatment group was correlated with reduced-ir TH terminals in the inner ear. These T-treatment induced changes in TH-ir fibers mimic the seasonal downregulation of dopamine in the midshipman inner ear and provide evidence that steroid hormone regulation of peripheral auditory sensitivity is mediated, in part, by dopamine.

The auditory system of the plainfin midshipman fish ( Porichthys notatus) is an important sensory system used to detect and encode biologically relevant acoustic stimuli important for survival and reproduction including social acoustic signals used for intraspecific communication. Previous work showed that hair cell (HC) density in the midshipman saccule increased seasonally with reproductive state and was concurrent with enhanced auditory saccular sensitivity in both females and type I males. Although reproductive state-dependent changes in HC density have been well characterized in the adult midshipman saccule, less is known about how the saccule changes during ontogeny. Here, we examined the ontogenetic development of the saccule in four relative sizes of midshipman (larvae, small juveniles, large juveniles, and nonreproductive adults) to determine whether the density, total number, and orientation patterns of saccular HCs change during ontogeny. In addition, we also examined whether the total number of HCs in the saccule differ from that of the utricle and lagena in nonreproductive adults. We found that HC density varied across developmental stage. The ontogenetic reduction in HC density was concurrent with an ontogenetic increase in macula area. The orientation pattern of saccular HCs was similar to the standard pattern previously described in other teleost fishes, and this pattern of HC orientation was retained during ontogeny. Lastly, the estimated number of saccular HCs increased with developmental stage from the smallest larvae (2,336 HCs) to the largest nonreproductive adult (145,717 HCs), and in nonreproductive adults estimated HC numbers were highest in the saccule (mean ± SD = 28,479 ± 4,809 HCs), intermediate in the utricle (mean ± SD = 11,008 ± 1,619 HCs) and lowest in the lagena (mean ± SD = 4,560 ± 769 HCs).

Investigators working with fish bioacoustics used to refer to fishes that have a narrow hearing bandwidth and poor sensitivity as “hearing generalists” (or “non-specialists”), while fishes that could detect a wider hearing bandwidth and had greater sensitivity were referred to as specialists. However, as more was learned about fish hearing mechanism and capacities, these terms became hard to apply, since it was clear there were gradations in hearing capabilities. Popper and Fay, in a paper in Hearing Research in 2011, proposed that these terms be dropped because of the gradation. While this was widely accepted by investigators, it is now apparent that the lack of relatively concise terminology for fish hearing capabilities makes it hard to discuss fish hearing. Thus, in this paper we resurrect the terms specialist and non-specialist but use them with modifiers to express the specific structure of function that is considered a specialization. Moreover, this resurrection recognizes that hearing specializations in fishes may not only be related to increased bandwidth and/or sensitivity, but to other, perhaps more important, aspects of hearing such as sound source localization, discrimination between sounds, and detection of sounds in the presence of masking signals.

The plainfin midshipman, Porichthys notatus, is a soniferous marine teleost fish that generates acoustic signals for intraspecific social communication. Nocturnally active males and females rely on their auditory sense to detect and locate vocally active conspecifics during social behaviors. Previous work showed that the midshipman inner ear saccule and lagena are highly adapted to detect and encode socially relevant acoustic stimuli, but the auditory sensitivity and function of the midshipman utricle remain largely unknown. Here, we characterize the auditory evoked potentials from hair cells in the utricle of non-reproductive type I males and test the hypothesis that the midshipman utricle is sensitive to behaviorally-relevant acoustic stimuli. Hair cell potentials were recorded from the rostral, medial and caudal regions of the utricle in response to pure tone stimuli presented by an underwater speaker. We show that the utricle is highly sensitive to particle motion stimuli produced by an underwater speaker positioned in the horizontal plane. Utricular potentials were recorded across a broad range of frequencies with lowest particle acceleration (dB re: 1 ms−2) thresholds occurring at 105 Hz (lowest frequency tested; mean threshold=-32 dB re: 1 ms−2) and highest thresholds at 605 to 1005 Hz (mean threshold range=−5 to −4 dB re: 1 ms−2). The high gain and broadband frequency sensitivity of the utricle suggests that it likely serves a primary auditory function and is well suited to detect conspecific vocalizations including broadband agonistic signals and the multiharmonic advertisement calls produce by reproductive type I males.

The plainfin midshipman fish (Porichthys notatus) is an established model for investigating acoustic communication because the reproductive success of this species is dependent on the production and reception of social acoustic signals. Previous work showed female midshipman have swim bladders with rostral horn-like extensions that project close to the saccule and lagena, while nesting (type I) males lack such rostral swim bladder extensions. The relative close proximity of the swim bladder to the lagena should increase auditory sensitivity to sound pressure and higher frequencies. Here, we test the hypothesis that the swim bladder of female midshipman enhances lagenar sensitivity to sound pressure and higher frequencies. Evoked potentials were recorded from auditory hair cell receptors in the lagena in reproductive females with intact (control condition) and removed (treated condition) swim bladders while pure tone stimuli (85–1005 Hz) were presented by an underwater speaker. Females with intact swim bladders had auditory thresholds 3–6 dB lower than females without swim bladders over a range of frequencies from 85–405 Hz. At frequencies from 545–1005 Hz, only females with intact swim bladders had measurable auditory thresholds (150–153 dB re: 1 µPa). The higher percentage of evoked lagenar potentials recorded in control females at frequencies>505 Hz indicates that the swim bladder extends the bandwidth of detectable frequencies. These findings reveal that the swim bladders in female midshipman can enhance lagenar sensitivity to sound pressure and higher frequencies, which maybe important for the detection of behaviorally relevant social signals.

While the peripheral auditory system of fish has been well studied, less is known about how the fish’s brain and central auditory system process complex social acoustic signals. The plainfin midshipman fish, Porichthys notatus, has become a good species for investigating the neural basis of acoustic communication because the production and reception of acoustic signals is paramount for this species’ reproductive success. Nesting males produce long-duration advertisement calls that females detect and localize among the noise in the intertidal zone to successfully find mates and spawn. How female midshipman are able to discriminate male advertisement calls from environmental noise and other acoustic stimuli is unknown. Using the immediate early gene product cFos as a marker for neural activity, we quantified neural activation of the ascending auditory pathway in female midshipman exposed to conspecific advertisement calls, heterospecific white seabass calls, or ambient environment noise. We hypothesized that auditory hindbrain nuclei would be activated by general acoustic stimuli (ambient noise and other biotic acoustic stimuli) whereas auditory neurons in the midbrain and forebrain would be selectively activated by conspecific advertisement calls. We show that neural activation in two regions of the auditory hindbrain, i.e., the rostral intermediate division of the descending octaval nucleus and the ventral division of the secondary octaval nucleus, did not differ via cFos immunoreactive (cFos-ir) activity when exposed to different acoustic stimuli. In contrast, female midshipman exposed to conspecific advertisement calls showed greater cFos-ir in the nucleus centralis of the midbrain torus semicircularis compared to fish exposed only to ambient noise. No difference in cFos-ir was observed in the torus semicircularis of animals exposed to conspecific versus heterospecific calls. However, cFos-ir was greater in two forebrain structures that receive auditory input, i.e., the central posterior nucleus of the thalamus and the anterior tuberal hypothalamus, when exposed to conspecific calls versus either ambient noise or heterospecific calls. Our results suggest that higher-order neurons in the female midshipman midbrain torus semicircularis, thalamic central posterior nucleus, and hypothalamic anterior tuberal nucleus may be necessary for the discrimination of complex social acoustic signals. Furthermore, neurons in the central posterior and anterior tuberal nuclei are differentially activated by exposure to conspecific versus other acoustic stimuli.

The plainfin midshipman fish, Porichthys notatus, is a nocturnal marine teleost that uses social acoustic signals for communication during the breeding season. Nesting type I males produce multiharmonic advertisement calls by contracting their swim bladder sonic muscles to attract females for courtship and spawning while subsequently attracting cuckholding type II males. Here, we report intra- and intersexual dimorphisms of the swim bladder in a vocal teleost fish and detail the swim bladder dimorphisms in the three sexual phenotypes (females, type I and II males) of plainfin midshipman fish. Micro-computerized tomography revealed that females and type II males have prominent, horn-like rostral swim bladder extensions that project toward the inner ear end organs (saccule, lagena, and utricle). The rostral swim bladder extensions were longer, and the distance between these swim bladder extensions and each inner-ear end organ type was significantly shorter in both females and type II males compared to that in type I males. Our results revealed that the normalized swim bladder length of females and type II males was longer than that in type I males while there was no difference in normalized swim bladder width among the three sexual phenotypes. We predict that these intrasexual and intersexual differences in swim bladder morphology among midshipman sexual phenotypes will afford greater sound pressure sensitivity and higher frequency detection in females and type II males and facilitate the detection and localization of conspecifics in shallow water environments, like those in which midshipman breed and nest.

Adult female and nesting (type I) male midshipman fish (Porichthys notatus) exhibit an adaptive form of auditory plasticity for the enhanced detection of social acoustic signals. Whether this adaptive plasticity also occurs in “sneaker” type II males is unknown. Here, we characterize auditory-evoked potentials recorded from hair cells in the saccule of reproductive and non-reproductive “sneaker” type II male midshipman to determine whether this sexual phenotype exhibits seasonal, reproductive state-dependent changes in auditory sensitivity and frequency response to behaviorally relevant auditory stimuli. Saccular potentials were recorded from the middle and caudal region of the saccule while sound was presented via an underwater speaker. Our results indicate saccular hair cells from reproductive type II males had thresholds based on measures of sound pressure and acceleration (re. 1 µPa and 1 ms−2, respectively) that were ~8–21 dB lower than non-reproductive type II males across a broad range of frequencies, which include the dominant higher frequencies in type I male vocalizations. This increase in type II auditory sensitivity may potentially facilitate eavesdropping by sneaker males and their assessment of vocal type I males for the selection of cuckoldry sites during the breeding season.

The plainfin midshipman (Porichthys notatus) is an acoustically communicative teleost fish. Here, we evaluated auditory evoked potentials (AEPs) in reproductive female midshipman exposed to tones at or near dominant frequencies of the male midshipman advertisement call. An initial series of experiments characterized AEPs at behaviorally relevant suprathreshold sound levels (130–140 dB SPL re. 1 µPa). AEPs decreased in magnitude with increasing stimulus frequency and featured a stereotyped component at twice the stimulus frequency. Recording electrode position was varied systematically and found to affect AEP magnitude and phase characteristics. Later experiments employed stimuli of a single frequency to evaluate contributions of the saccule to the AEP, with particular attention to the effects of sound source azimuth on AEP amplitude. Unilateral excision of saccular otoliths (sagittae) decreased AEP amplitude; unexpectedly, decreases differed for right versus left otolith excision. A final set of experiments manipulated the sound pressure-responsive swim bladder. Swim bladder excision further reduced the magnitude of AEP responses, effectively eliminating responses at the standard test intensity (130 dB SPL) in some animals. Higher-intensity stimulation yielded response minima at forward azimuths ipsilateral to the excised sagitta, but average cross-azimuth modulation generally remained slight. Collectively, the data underscore that electrode position is an essential variable to control in fish AEP studies and suggest that in female midshipman: (1) the saccule contributes to the AEP, but its directionality as indexed by the AEP is limited, (2) a left–right auditory asymmetry may exist and (3) the swim bladder provides gain in auditory sensitivity that may be important for advertisement call detection and phonotaxis.

The plainfin midshipman fish, Porichthys notatus, is a seasonally breeding, nocturnal marine teleost fish that produces acoustic signals for intraspecific social communication. Females rely on audition to detect and locate “singing” males that produce multiharmonic advertisement calls in the shallow-water, intertidal breeding environments. Previous work showed that females possess sexually-dimorphic, horn-like rostral swim bladder extensions that extend toward the primary auditory end organs, the saccule and lagena. Here, we test the hypothesis that the rostral swim bladder extensions in females increase auditory sensitivity to sound pressure and higher frequencies, which potentially could enhance mate detection and localization in shallow water habitats. We recorded the auditory evoked potentials that originated from hair cell receptors in the saccule of control females with intact swim bladders and compared them with that from treated females (swim bladders removed) and type I males (intact swim bladders lacking rostral extensions). Saccular potentials were recorded from hair cell populations in vivo while behaviorally relevant pure tone stimuli (75-1005 Hz) were presented by an underwater speaker. Results indicate that control females were approximately 5-11 dB (re: 1 µPa) more sensitive to sound pressure than treated females and type I males at the frequencies tested. A higher percentage of the evoked saccular potentials were recorded from control females at frequencies >305 Hz than from treated females and type I males. This enhanced sensitivity in females to sound pressure and higher frequencies may facilitate the acquisition of auditory information needed for conspecific localization and mate choice decisions during the breeding season.

Summary Dopamine is integral to attentional and motivational processes, but studies are largely restricted to the central nervous system. In mammals [ 1 , 2 ] and fishes [ 3 , 4 ], central dopaminergic neurons project to the inner ear and could modulate acoustic signals at the earliest stages of processing. Studies in rodents show dopamine inhibits cochlear afferent neurons and protects against noise-induced acoustic injury [ 5 , 6 , 7 , 8 , 9 , 10 ]. However, other functions for inner ear dopamine have not been investigated, and the effect of dopamine on peripheral auditory processing in non-mammalians remains unknown [ 11 , 12 ]. Insights could be gained by studies conducted in the context of intraspecific acoustic communication. We present evidence from a vocal fish linking reproductive-state-dependent changes in auditory sensitivity with seasonal changes in the dopaminergic efferent system in the saccule, their primary organ of hearing. Plainfin midshipman (Porichthys notatus) migrate from deep-water winter habitats to the intertidal zone in the summer to breed. Nesting males produce nocturnal vocalizations to attract females [ 13 ]. Both sexes undergo seasonal enhancement of hearing sensitivity at the level of the hair cell [ 14 , 15 , 16 ], increasing the likelihood of detecting conspecific signals [ 17 , 18 ]. Importantly, reproductive females concurrently have reduced dopaminergic input to the saccule [ 19 ]. Here, we show that dopamine decreases saccule auditory sensitivity via a D2-like receptor. Saccule D2a receptor expression is reduced in the summer and correlates with sensitivity within and across seasons. We propose that reproductive-state-dependent changes to the dopaminergic efferent system provide a release of inhibition in the saccule, enhancing peripheral encoding of social-acoustic signals.

Age-related hearing loss (ARHL), also known as presbycusis, is a widespread and debilitating condition impacting many older adults. Conventionally, researchers utilize mammalian model systems or human cadaveric tissue to study ARHL pathology. Recently, the zebrafish has become an effective and tractable model system for a wide variety of genetic and environmental auditory insults, but little is known about the incidence or extent of ARHL in zebrafish and other non-mammalian models. Here, we evaluated whether zebrafish exhibit age-related loss in auditory sensitivity. The auditory sensitivity of adult wild-type zebrafish (AB/WIK strain) from three adult age subgroups (13-month, 20-month, and 37-month) was characterized using the auditory evoked potential (AEP) recording technique. AEPs were elicited using pure tone stimuli (115–4500 Hz) presented via an underwater loudspeaker and recorded using shielded subdermal metal electrodes. Based on measures of sound pressure and particle acceleration, the mean AEP thresholds of 37-month-old fish [mean sound pressure level (SPL) = 122.2 dB ± 2.2 dB SE re: 1 μPa; mean particle acceleration level (PAL) = -27.5 ± 2.3 dB SE re: 1 ms−2] were approximately 9 dB higher than that of 20-month-old fish [(mean SPL = 113.1 ± 2.7 dB SE re: 1 μPa; mean PAL = -37.2 ± 2.8 dB re: 1 ms−2; p = 0.007)] and 6 dB higher than that of 13-month-old fish [(mean SPL = 116.3 ± 2.5 dB SE re: 1 μPa; mean PAL = -34.1 ± 2.6 dB SE re: 1 ms−2; p = 0.052)]. Lowest AEP thresholds for all three age groups were generally between 800 Hz and 1850 Hz, with no evidence for frequency-specific age-related loss. Our results suggest that zebrafish undergo age-related loss in auditory sensitivity, but the form and magnitude of loss is markedly different than in mammals, including humans. Future work is needed to further describe the incidence and extent of ARHL across vertebrate groups and to determine which, if any, ARHL mechanisms may be conserved across vertebrates to support meaningful comparative/translational studies.

The plainfin midshipman fish (Porichthys notatus) is a species of marine teleost that produces acoustic signals that are important for mediating social behavior. The auditory sensitivity of the saccule is well established in this species, but the sensitivity and function of the midshipman’s putative auditory lagena are unknown. Here, we characterize the auditory-evoked potentials from hair cells in the lagena of reproductive type I males to determine the frequency response and auditory sensitivity of the lagena to behaviorally relevant acoustic stimuli. Lagenar potentials were recorded from the caudal and medial region of the lagena, while acoustic stimuli were presented by an underwater speaker. Our results indicate that the midshipman lagena has a similar low-frequency sensitivity to that of the midshipman saccule based on sound pressure and acceleration (re: 1 µPa and 1 ms(−2), respectively), but the thresholds of the lagena were higher across all frequencies tested. The relatively high auditory thresholds of the lagena may be important for encoding high levels of behaviorally relevant acoustic stimuli when close to a sound source.

Seasonal changes in reproductive-related vocal behavior are widespread among fishes. This review highlights recent studies of the vocal plainfin midshipman fish, Porichthys notatus, a neuroethological model system used for the past two decades to explore neural and endocrine mechanisms of vocal-acoustic social behaviors shared with tetrapods. Integrative approaches combining behavior, neurophysiology, neuropharmacology, neuroanatomy, and gene expression methodologies have taken advantage of simple, stereotyped and easily quantifiable behaviors controlled by discrete neural networks in this model system to enable discoveries such as the first demonstration of adaptive seasonal plasticity in the auditory periphery of a vertebrate as well as rapid steroid and neuropeptide effects on vocal physiology and behavior. This simple model system has now revealed cellular and molecular mechanisms underlying seasonal and steroid-driven auditory and vocal plasticity in the vertebrate brain.

Little is known regarding the coordination of audition with decision-making and subsequent motor responses that initiate social behavior including mate localization during courtship. Using the midshipman fish model, we tested the hypothesis that the time spent by females attending and responding to the advertisement call is correlated with the activation of a specific subset of catecholaminergic (CA) and social decision-making network (SDM) nuclei underlying auditory- driven sexual motivation. In addition, we quantified the relationship of neural activation between CA and SDM nuclei in all responders with the goal of providing a map of functional connectivity of the circuitry underlying a motivated state responsive to acoustic cues during mate localization. In order to make a baseline qualitative comparison of this functional brain map to unmotivated females, we made a similar correlative comparison of brain activation in females who were unresponsive to the advertisement call playback. Our results support an important role for dopaminergic neurons in the periventricular posterior tuberculum and ventral thalamus, putative A11 and A13 tetrapod homologues, respectively, as well as the posterior parvocellular preoptic area and dorsomedial telencephalon, (laterobasal amygdala homologue) in auditory attention and appetitive sexual behavior in fishes. These findings may also offer insights into the function of these highly conserved nuclei in the context of auditory-driven reproductive social behavior across vertebrates.

Overexposure to loud noise is known to lead to deficits in auditory sensitivity and perception. We studied the effects of noise exposure on sensorimotor behaviors of larval (5–7 days post-fertilization) zebrafish (Danio rerio), particularly the auditory-evoked startle response and hearing sensitivity to acoustic startle stimuli. We observed a temporary 10–15 dB decrease in startle response threshold after 18 h of flat-spectrum noise exposure at 20 dB re·1 ms(−2). Larval zebrafish also exhibited decreased habituation to startle-inducing stimuli following noise exposure. The noise-induced sensitization was not due to changes in absolute hearing thresholds, but was specific to the auditory-evoked escape responses. The observed noise-induced sensitization was disrupted by AMPA receptor blockade using DNQX, but not NMDA receptor blockade. Together, these experiments suggest a complex effect of noise exposure on the neural circuits mediating auditory-evoked behaviors in larval zebrafish.

Although the neuroanatomical distribution of catecholaminergic (CA) neurons has been well documented across all vertebrate classes, few studies have examined CA connectivity to physiologically and anatomically identified neural circuitry that controls behavior. The goal of this study was to characterize CA distribution in the brain and inner ear of the plainfin midshipman fish (Porichthys notatus) with particular emphasis on their relationship with anatomically labeled circuitry that both produces and encodes social acoustic signals in this species. Neurobiotin labeling of the main auditory end organ, the saccule, combined with tyrosine hydroxylase immunofluorescence (TH-ir) revealed a strong CA innervation of both the peripheral and central auditory system. Diencephalic TH-ir neurons in the periventricular posterior tuberculum, known to be dopaminergic, send ascending projections to the ventral telencephalon and prominent descending projections to vocal-acoustic integration sites, notably the hindbrain octavolateralis efferent nucleus, as well as onto the base of hair cells in the saccule via nerve VIII. Neurobiotin backfills of the vocal nerve in combination with TH-ir revealed CA terminals on all components of the vocal pattern generator, which appears to largely originate from local TH-ir neurons but may include input from diencephalic projections as well. This study provides strong neuroanatomical evidence that catecholamines are important modulators of both auditory and vocal circuitry and acoustic-driven social behavior in midshipman fish. This demonstration of TH-ir terminals in the main end organ of hearing in a nonmammalian vertebrate suggests a conserved and important anatomical and functional role for dopamine in normal audition.

In humans, excessive noise exposure from occupational or recreational sources causes permanent hearing loss. Similarly, exposure to underwater anthropogenic noise can cause hearing loss in aquatic organisms, including fish. While fish can recover from noise-induced hearing loss, underwater noise exposure can cause behavioral changes that reduce organismal fitness. In all vertebrates, acoustic trauma can cause damage to sensory hair cells. To better study the effects of noise on hair cells, we have developed a noise exposure system that uses broadband sound to damage hair cells of the inner ear and lateral line of larval zebrafish. Acoustic over-exposure kills hair cells in an intensity- and time-dependent manner, with maximum hair cell damage observed 72 hours after noise exposure. This time course is consistent with mammalian studies, where hair cell death occurs days to weeks after noise exposure. Other features of acoustic trauma are also conserved between zebrafish and mammals, including activation of apoptotic signaling cascades and changes in hair cell-afferent synapses. These studies demonstrate that larval zebrafish are a tractable new model for studies of noise-induced hair cell death. However, our acoustic trauma system could also be used in other species, allowing for new studies of underwater noise in larval fishes.

The plainfin midshipman fish (Porichthys notatus) is a nocturnal, seasonally breeding, intertidal-nesting teleost fish that produces social acoustic signals for intraspecific communication. Type I or “nesting” males produce agonistic and reproductive-related acoustic signals including a multiharmonic advertisement call during the summer breeding season. Previous work showed that type I male auditory sensitivity of the saccule, the primary midshipman auditory end organ, changes seasonally with reproductive state such that reproductive males become more sensitive and better suited than nonreproductive males to detect the dominant frequencies contained within type I vocalizations. Here, we examine whether reproductive type I males also exhibit reproductive-state dependent changes in hair cell (HC) density in the three putative auditory end organs (saccule, lagena, and utricle). We show that saccular HC density was greater in reproductive type I males compared to nonreproductive type I males, and that the increase in HC density occurs throughout the saccular epithelium in both the central and marginal epithelia regions. We also show as saccular HC density increases there is a concurrent decrease in saccular support cell (SC) density in reproductive type I males with no overall change in total cell density (i.e., HC + SC). In contrast, we did not observe any seasonal changes in HC density in the utricle or lagena between nonreproductive and reproductive type I males. In addition, we compare the saccular HC densities in reproductive type I males with that of reproductive females and show that females have greater saccular HC densities, which suggest a sexually dimorphic difference in HC receptor density between the two sexual phenotypes, at least during the summer breeding season.

SUMMARYZebrafish (Danio rerio) have become a valuable model for investigating the molecular genetics and development of the inner ear in vertebrates. In this study, we employed a prepulse inhibition (PPI) paradigm to assess hearing in larval wild-type (AB) zebrafish during early development at 5–6 days post-fertilization (d.p.f.). We measured the PPI of the acoustic startle response in zebrafish using a 1-dimensional shaker that simulated the particle motion component of sound along the fish's dorsoventral axis. The thresholds to startle-inducing stimuli were determined in 5–6 d.p.f. zebrafish, and their hearing sensitivity was then characterized using the thresholds of prepulse tone stimuli (90–1200 Hz) that inhibited the acoustic startle response to a reliable startle stimulus (820 Hz at 20 dB re. 1 m s−2). Hearing thresholds were defined as the minimum prepulse tone level required to significantly reduce the startle response probability compared with the baseline (no-prepulse) condition. Larval zebrafish showed greatest auditory sensitivity from 90 to 310 Hz with corresponding mean thresholds of −19 to −10 dB re. 1 m s−2, respectively. Hearing thresholds of prepulse tones were considerably lower than previously predicted by startle response assays. The PPI assay was also used to investigate the relative contribution of the lateral line to the detection of acoustic stimuli. After aminoglycoside-induced neuromast hair-cell ablation, we found no difference in PPI thresholds between treated and control fish. We propose that this PPI assay can be used to screen for novel zebrafish hearing mutants and to investigate the ontogeny of hearing in zebrafish and other fishes.

Zebrafish are increasingly used in auditory studies, in part due to the development of several transgenic lines that express hair cell-specific fluorescent proteins. However, it is largely unknown how transgene expression influences auditory phenotype. We previously observed reduced auditory sensitivity in adult Brn3c:mGFP transgenic zebrafish, which express membrane-bound green fluorescent protein (GFP) in sensory hair cells. Here, we examine the auditory sensitivity of zebrafish from multiple transgenic and background strains. We recorded auditory evoked potentials in adult animals and observed significantly higher auditory thresholds in three lines that express hair cell-specific GFP. There was no obvious correlation between hair cell density and auditory thresholds, suggesting that reduced sensitivity was not due to a reduction in hair cell density. FM1-43 uptake was reduced in Brn3c:mGFP fish but not in other lines, suggesting that a mechanotransduction defect may be responsible for the auditory phenotype in Brn3c animals, but that alternate mechanisms underlie the increased AEP thresholds in other lines. We found reduced prepulse inhibition (a measure of auditory-evoked behavior) in larval Brn3c animals, suggesting that auditory defects develop early in this line. We also found significant differences in auditory sensitivity between adults of different background strains, akin to strain differences observed in mouse models of auditory function. Our results suggest that researchers should exercise caution when selecting an appropriate zebrafish transgenic or background strain for auditory studies.

Sounds provide fishes with important information used to mediate behaviors such as predator avoidance, prey detection, and social communication. How we measure auditory capabilities in fishes, therefore, has crucial implications for interpreting how individual species use acoustic information in their natural habitat. Recent analyses have highlighted differences between behavioral and electrophysiologically determined hearing thresholds, but less is known about how physiological measures at different auditory processing levels compare within a single species. Here we provide one of the first comparisons of auditory threshold curves determined by different recording methods in a single fish species, the soniferous Hawaiian sergeant fish Abudefduf abdominalis, and review past studies on representative fish species with tuning curves determined by different methods. The Hawaiian sergeant is a colonial benthic-spawning damselfish (Pomacentridae) that produces low-frequency, low-intensity sounds associated with reproductive and agonistic behaviors. We compared saccular potentials, auditory evoked potentials (AEP), and single neuron recordings from acoustic nuclei of the hindbrain and midbrain torus semicircularis. We found that hearing thresholds were lowest at low frequencies (~75-300 Hz) for all methods, which matches the spectral components of sounds produced by this species. However, thresholds at best frequency determined via single cell recordings were ~15-25 dB lower than those measured by AEP and saccular potential techniques. While none of these physiological techniques gives us a true measure of the auditory "perceptual" abilities of a naturally behaving fish, this study highlights that different methodologies can reveal similar detectable range of frequencies for a given species, but absolute hearing sensitivity may vary considerably.

Fish represent the largest group of vertebrates and display the greatest diversity of auditory structures. However, studies addressing how the form and function of the auditory system change during development to enhance perception of the acoustic environment are rather sparse in this taxon compared to other vertebrate groups. An ontogenetic perspective of the auditory system in fishes provides a readily testable framework for understanding structure-function relationships. Additionally, studying ancestral models such as fish can convey valuable comparable information across vertebrates, as early developmental events are often evolutionary conserved. This chapter reviews the literature on the morphological development of the fish auditory system, with particular focus on the inner ear structures that evolve from an otic placode during early embryonic development and then continue to undergo differentiation and maturation in the postembryonic phase. Moreover, the chapter provides a systematic overview of how auditory sensitivity develops during ontogeny. Although most studies indicate a developmental improvement in auditory sensitivity, there is considerably species-specific variation. Lastly, the paucity of information and literature concerning the development of auditory capabilities for social communication in fishes is also discussed. Further investigation on the development of structure and function of the fish auditory system is recommended in order to obtain a deeper understanding of how ontogenetic morphological changes in the auditory pathway relate to modifications in acoustic reception, auditory processing, and the capacity to communicate acoustically.

This paper reviews recent studies on the effects of hatchery rearing on the auditory and lateral line systems of salmonid fishes. Major conclusions are that (1) hatchery-reared juveniles exhibit abnormal lateral line morphology (relative to wild-origin conspecifics), suggesting that the hatchery environment affects lateral line structure, perhaps due to differences in the hydrodynamic conditions of hatcheries versus natural rearing environments, and (2) hatchery-reared salmonids have a high proportion of abnormal otoliths, a condition associated with reduced auditory sensitivity and suggestive of inner ear dysfunction.

Auditory evoked potentials (AEPs) have become popular for estimating hearing thresholds and audiograms. What is the utility of these measurements? How do AEP audiograms compare with behavioral audiograms? In general, AEP measurements for fishes and marine mammals often underestimate behavioral thresholds, but comparisons are especially complicated when the AEP and behavioral measures are obtained under different acoustic conditions. There is no single representative relationship between AEP and behavioral audiograms and these audiograms should not be considered equivalent. We suggest that the most valuable comparisons are those made by the same researcher using similar acoustic conditions for both measurements.

Evidence suggests that the capacity for sound source localization is common to mammals, birds, reptiles, and amphibians, but surprisingly it is not known whether fish locate sound sources in the same manner (e.g., combining binaural and monaural cues) or what computational strategies they use for successful source localization. Directional hearing and sound source localization in fishes continues to be important topics in neuroethology and in the hearing sciences, but the empirical and theoretical work on these topics have been contradictory and obscure for decades. This chapter reviews the previous behavioral work on directional hearing and sound source localization in fishes including the most recent experiments on sound source localization by the plainfin midshipman fish (Porichthys notatus), which has proven to be an exceptional species for fish studies of sound localization. In addition, the theoretical models of directional hearing and sound source localization for fishes are reviewed including a new model that uses a time-averaged intensity approach for source localization that has wide applicability with regard to source type, acoustic environment, and time waveform.

On May 25, 2013, a special symposium was held at the Mote Marine Laboratory in Sarasota, FL to honor the outstanding careers of Drs. Richard R. Fay and Arthur N. Popper, a “dynamic duo” of scientists who were pioneers in the field of contemporary fish hearing and bioacoustics. The present article details the research, academic life, and “other side” of Richard Rozzell Fay, a most interesting man of science who is known to all as a kind, gentle, wise, and introspective scientist.

The plainfin midshipman fish (Porichthys notatus) is a well-studied model to understand the neural and endocrine mechanisms underlying vocal-acoustic communication across vertebrates. It is well established that steroid hormones such as estrogen drive seasonal peripheral auditory plasticity in female Porichthys in order to better encode the male’s advertisement call. However, little is known of the neural substrates that underlie the motivation and coordinated behavioral response to auditory social signals. Catecholamines, which include dopamine and noradrenaline, are good candidates for this function, as they are thought to modulate the salience of and reinforce appropriate behavior to socially relevant stimuli. This chapter summarizes our recent studies which aimed to characterize catecholamine innervation in the central and peripheral auditory system of Porichthys as well as test the hypotheses that innervation of the auditory system is seasonally plastic and catecholaminergic neurons are activated in response to conspecific vocalizations. Of particular significance is the discovery of direct dopaminergic innervation of the saccule, the main hearing end organ, by neurons in the diencephalon, which also robustly innervate the cholinergic auditory efferent nucleus in the hindbrain. Seasonal changes in dopamine innervation in both these areas appear dependent on reproductive state in females and may ultimately function to modulate the sensitivity of the peripheral auditory system as an adaptation to the seasonally changing soundscape. Diencephalic dopaminergic neurons are indeed active in response to exposure to midshipman vocalizations and are in a perfect position to integrate the detection and appropriate motor response to conspecific acoustic signals for successful reproduction.

Behavioral methods have been critical in the study of auditory perception and discrimination in fishes. In this chapter, we review some of the common methods used in fish psychoacoustics. We discuss associative methods, such as operant, avoidance, and classical conditioning, and their use in constructing audiograms, measuring frequency selectivity, and auditory stream segregation. We also discuss the measurement of innate behavioral responses, such as the acoustic startle response (ASR), prepulse inhibition (PPI), and phonotaxis, and their use in the assessment of fish hearing to determine auditory thresholds and in the testing of mechanisms for sound source localization. For each psychoacoustic method, we provide examples of their use and discuss the parameters and situations where such methods can be best utilized. In the case of the ASR, we show how this method can be used to construct and compare audiograms between two species of larval fishes, the three-spined stickleback (Gasterosteus aculeatus) and the zebrafish (Danio rerio). We also discuss considerations for experimental design with respect to stimulus presentation and threshold criteria and how these techniques can be used in future studies to investigate auditory perception in fishes.

Fishes, the most species-rich group of living vertebrates, present remarkable opportunities to investigate neural mechanisms underlying the sense of hearing. While the sound-producing and hearing abilities of fishes have long been known, it was not until the turn of this century that the influence of hormones on audition was first reported for fish. This discovery has led to numerous studies in multiple species using a range of methodological approaches to investigate underlying cellular and molecular mechanisms. Here, we review these findings within behavioral, hormonal, ecological, and evolutionary contexts. As shown, fishes share a large suite of behavioral and neurohormonal characters with tetrapods and offer distinct advantages as experimental models, in part, because of easy accessibility to the peripheral and central auditory systems.

Van Trump et al. (2010) recently demonstrated that the aminoglycoside antibiotic gentamicin, previously thought to be selectively toxic to hair cells of the canal neuromasts (CN) of the fish lateral line system, is additionally toxic to hair cells of the superficial neuromasts (SN). The authors used the fluorescent vital dyes DASPEI (2-(4-(dimethylamino)styryl)-N-ethylpyridinium iodide) and FM1-43 ((n-(3-triethylammoniumpropyl)-4-(4-(dibutylamino)styryl) pyridinium dibromide) to visualize by fluorescence microscopy the survival of hair cells in the CN and SN of zebrafish (Danio rerio) and Mexican blind cave fish (Astyanax fasciatus) treated in a solution of 0.001% gentamicin sulfate for 24h. Extensive hair cell death was observed in populations of CN and SN of gentamicin-treated animals of both species compared to control animals. On the basis of this result, the authors concluded, (p. 49), “(1) That hair cells in the SNs of the lateral line can no longer be regarded as functionally resistant to gentamicin toxicity, (2) that this drug should therefore no longer be used as a pharmacological tool for selective blocking of CN, but not SN hair cells, and (3) that the conclusions of some previous studies need to be reevaluated.” Using an alternative approach to themselves reevaluate a 15-year-old assumption in the literature drawn from SEM imaging (e.g., Song et al., 1995), Van Trump et al. (2010) convincingly demonstrated that gentamicin in fact damages a high proportion of both SN and CN, similar in effect to another commonly used aminoglycoside, streptomycin (e.g., Kaus, 1987; Blaxter and Fuiman, 1989; Montgomery et al., 1997; Montgomery et al., 2003). Toward further caution in the interpretation of lateral line behavioral studies that have used aminoglycoside antibiotics to chemically ablate SN and/or CN without rigorous verification of treatment efficacy, we are inspired to share our own recent observations on limitations in the efficacy of both gentamicin and streptomycin in damaging the lateral line systems of three fish species.

The auditory system of the plainfin midshipman fish, Porichthys notatus, is an important sensory receiver system used to encode intraspecific social communication signals in adults, but the response properties and function of this receiver system in pre-adult stages are less known. In this study we examined the response properties of auditory-evoked potentials from the midshipman saccule, the main organ of hearing in this species, to determine whether the frequency response and auditory threshold of saccular hair cells to behaviorally relevant single tone stimuli change during ontogeny. Saccular potentials were recorded from three relative sizes of midshipman fish: small juveniles [1.9–3.1 cm standard length (SL), large juveniles (6.8–8.0 cm SL) and non-reproductive adults (9.0–22.6 cm SL)]. The auditory evoked potentials were recorded from the rostral, middle and caudal regions of the saccule while single tone stimuli (75–1,025 Hz) were presented via an underwater speaker. We show that the frequency response and auditory threshold of the midshipman saccule is established early in development and retained throughout ontogeny. We also show that saccular sensitivity to frequencies greater than 385 Hz increases with age/size and that the midshipman saccule of small and large juveniles, like that of non-reproductive adults, is best suited to detect low frequency sounds (

Excessive noise exposure damages sensory hair cells, leading to permanent hearing loss. Zebrafish are a highly tractable model that have advanced our understanding of drug-induced hair cell death, yet no comparable model exists for noise exposure research. We demonstrate the utility of zebrafish as model to increase understanding of hair cell damage from acoustic trauma and develop protective therapies. We created an acoustic trauma system using underwater cavitation to stimulate lateral line hair cells. We found that acoustic stimulation resulted in exposure time- and intensity-dependent lateral line and saccular hair cell damage that is maximal at 48–72 h post-trauma. The number of TUNEL+ lateral line hair cells increased 72 h post-exposure, whereas no increase was observed in TUNEL+ supporting cells, demonstrating that acoustic stimulation causes hair cell-specific damage. Lateral line hair cells damaged by acoustic stimulation regenerate within 3 d, consistent with prior regeneration studies utilizing ototoxic drugs. Acoustic stimulation-induced hair cell damage is attenuated by pharmacological inhibition of protein synthesis or caspase activation, suggesting a requirement for translation and activation of apoptotic signaling cascades. Surviving hair cells exposed to acoustic stimulation showed signs of synaptopathy, consistent with mammalian studies. Finally, we demonstrate the feasibility of this platform to identify compounds that prevent acoustic trauma by screening a small redox library for protective compounds. Our data suggest that acoustic stimulation results in lateral line hair cell damage consistent with acoustic trauma research in mammals, providing a highly tractable model for high-throughput genetic and drug discovery studies.

The plainfin midshipman fish (Porichthys notatus Girard, 1854) is a vocal species of batrachoidid fish that generates acoustic signals for intraspecific communication during social and reproductive activity and has become a good model for investigating the neural and endocrine mechanisms of vocal-acoustic communication. Reproductively active female plainfin midshipman fish use their auditory sense to detect and locate "singing" males, which produce a multiharmonic advertisement call to attract females for spawning. The seasonal onset of male advertisement calling in the midshipman fish coincides with an increase in the range of frequency sensitivity of the female's inner ear saccule, the main organ of hearing, thus leading to enhanced encoding of the dominant frequency components of male advertisement calls. Non-reproductive females treated with either testosterone or 17β-estradiol exhibit a dramatic increase in the inner ear's frequency sensitivity that mimics the reproductive female's auditory phenotype and leads to an increased detection of the male's advertisement call. This novel form of auditory plasticity provides an adaptable mechanism that enhances coupling between sender and receiver in vocal communication. This review focuses on recent evidence for seasonal reproductive-state and steroid-dependent plasticity of auditory frequency sensitivity in the peripheral auditory system of the midshipman fish. The potential steroid-dependent mechanism(s) that lead to this novel form of auditory and behavioral plasticity are also discussed.

Vocal differentiation is widely documented in birds and mammals but has been poorly investigated in other vertebrates, including fish, which represent the oldest extant vertebrate group. Neural circuitry controlling vocal behaviour is thought to have evolved from conserved brain areas that originated in fish, making this taxon key to understanding the evolution and development of the vertebrate vocal-auditory systems. This study examines ontogenetic changes in the vocal repertoire and whether vocal differentiation parallels auditory development in the Lusitanian toadfish Halobatrachus didactylus (Batrachoididae). This species exhibits a complex acoustic repertoire and is vocally active during early development. Vocalisations were recorded during social interactions for four size groups (fry: 25 cm, standard length). Auditory sensitivity of juveniles and adults was determined based on evoked potentials recorded from the inner ear saccule in response to pure tones of 75-945 Hz. We show an ontogenetic increment in the vocal repertoire from simple broadband-pulsed 'grunts' that later differentiate into four distinct vocalisations, including low-frequency amplitude-modulated 'boatwhistles'. Whereas fry emitted mostly single grunts, large juveniles exhibited vocalisations similar to the adult vocal repertoire. Saccular sensitivity revealed a three-fold enhancement at most frequencies tested from small to large juveniles; however, large juveniles were similar in sensitivity to adults. We provide the first clear evidence of ontogenetic vocal differentiation in fish, as previously described for higher vertebrates. Our results suggest a parallel development between the vocal motor pathway and the peripheral auditory system for acoustic social communication in fish. Fundo para o Desenvolvimento das Ciências e da Tecnologia, Macau S.A.R.; Fundaçao para a Ciência e a Tecnologia (FCT); Royalty Research Fund, USA

Acoustic communication is essential for the reproductive success of the plainfin midshipman fish (Porichthys notatus). During the breeding season, type I males use acoustic cues to advertise nest location to potential mates, creating an audible signal that attracts reproductive females. Type II (sneaker) males also likely use this social acoustic signal to find breeding pairs from which to steal fertilizations. Estrogen-induced changes in the auditory system of breeding females are thought to enhance neural encoding of the advertisement call, and recent anatomical data suggest the saccule (the main auditory end organ) as one possible target for this seasonal modulation. Here we describe saccular transcriptomes from all three sexual phenotypes (females, type I and II males) collected during the breeding season as a first step in understanding the mechanisms underlying sexual phenotype-specific and seasonal differences in auditory function. We used RNA-Seq on the Ion Torrent platform to create a combined transcriptome dataset containing over 79,000 assembled transcripts representing almost 9,000 unique annotated genes. These identified genes include several with known inner ear function and multiple steroid hormone receptors. Transcripts most closely matched to published genomes of nile tilapia and large yellow croaker, inconsistent with the phylogenetic relationship between these species but consistent with the importance of acoustic communication in their life-history strategies. We then compared the RNA-Seq results from the saccules of reproductive females with a separate transcriptome from the non-reproductive female phenotype and found over 700 differentially expressed transcripts, including members of the Wnt and Notch signaling pathways that mediate cell proliferation and hair cell addition in the inner ear. These data constitute a valuable resource for furthering our understanding of the molecular basis for peripheral auditory function as well as a range of future midshipman and cross-species comparative studies of the auditory periphery.

SUMMARY The auditory system of adult midshipman fish Porichthys notatusGirard is an important sensory receiver system used during intraspecific social communication to encode conspecific vocalizations, but the response properties and function of this system in the pre-adult stages are unknown. Midshipman fish, like other teleosts, use the saccule as the main acoustic end organ of the inner ear. In this study, we examined the discharge properties and the frequency response dynamics of auditory saccular afferent neurons in pre-adult midshipman (∼4–12 months of age) to determine whether encoding of auditory information, inclusive of conspecific vocalizations,changes across life history stages. Extracellular single unit recordings were made from saccular afferents while sound was presented via an underwater speaker. Comparisons with adult data show that the resting discharge rate and auditory threshold sensitivity increased with age/size,while temporal encoding of frequency did not show any significant shifts. The results indicate that the saccular afferents of juveniles, like those of non-reproductive adults, are best adapted to temporally encode the low frequency components (≤100 Hz) of midshipman vocalizations. This report represents the first in vivo investigation of age-related changes in the encoding properties of individual auditory neurons for any fish species.

For seasonally breeding vertebrates, reproductive cycling is often coupled with changes in vocalizations that function in courtship and territoriality. Less is known about changes in auditory sensitivity to those vocalizations. Here, we show that nonreproductive female midshipman fish treated with either testosterone or 17β-estradiol exhibit an increase in the degree of temporal encoding of the frequency content of male vocalizations by the inner ear that mimics the reproductive female's auditory phenotype. This sensory plasticity provides an adaptable mechanism that enhances coupling between sender and receiver in vocal communication.

This study characterized the seasonal variation of the steroid hormones testosterone (T), 11-ketotestosterone (11-KT), 17beta-estradiol (E2), and cortisol (F) as they relate to the gonadal development and reproductive behavior of the plainfin midshipman fish, Porichthys notatus. The plainfin midshipman is a deep-water teleost that seasonally migrates into the shallow intertidal zone where type I, or "singing," males build nests, acoustically court and spawn with females. The gonadosomatic index and plasma steroid levels were measured from adult type I males and females collected over four time periods (non-reproductive, pre-nesting, nesting, and post-nesting) that corresponded to seasonal fluctuations in midshipman reproductive biology and behavior. Among type I males, plasma levels of T and 11-KT were low during the winter non-reproductive period, gradually increased during seasonal recrudescence of the testes in the spring pre-nesting period, and then peaked at the beginning of the summer nesting period. In the latter half of the nesting period and during the fall post-nesting period, plasma levels of T and 11-KT were low or non-detectable. Low, detectable levels of E2 were also found in the plasma of 50% or more type I males during every seasonal period except the winter non-reproductive period. Among females, plasma levels of T and E2 were low throughout the year but briefly peaked in April during the spring pre-nesting period when ovaries underwent seasonal recrudescence. Plasma F levels were correlated with collection depth and were lower in males than females when fish were collected deeper than 120 m. The sex-specific peaks of steroid hormone levels in male and female midshipman may serve differential functions related to the physiology, reproductive behavior, and vocal communication of this species.

Despite identification of multiple factors mediating salmon survival, significant disparities in survival-to-adulthood among hatchery- versus wild-origin juveniles persist. In the present report, we explore the hypothesis that hatchery-reared juveniles might exhibit morphological defects in vulnerable mechanosensory systems prior to release from the hatchery, potentiating reduced survival after release. Juvenile steelhead (Oncorhynchus mykiss) from two different hatcheries were compared to wild-origin juveniles on several morphological traits including lateral line structure, otolith composition (a proxy for auditory function), and brain weight. Wild juveniles were found to possess significantly more superficial lateral line neuromasts than hatchery-reared juveniles, although the number of hair cells within individual neuromasts was not significantly different across groups. Wild juveniles were also found to possess primarily normal, aragonite-containing otoliths, while hatchery-reared juveniles possessed a high proportion of crystallized (vaterite) otoliths. Finally, wild juveniles were found to have significantly larger brains than hatchery-reared juveniles. These differences together predict reduced sensitivity to biologically important hydrodynamic and acoustic signals from natural biotic (predator, prey, conspecific) and abiotic (turbulent flow, current) sources among hatchery-reared steelhead, in turn predicting reduced survival fitness after release. Physiological and behavioral studies are required to establish the functional significance of these morphological differences.

Sharks are practically legendary for their sensory capabilities, with some of this reputation deserved and some exaggerated. Accounts of sharks being able to smell or hear a single sh from miles away may be sh stories, but controlled measurements of elasmobranch sensory function have revealed that these animals possess an exquisite array of sensory systems for detecting prey and conspecics, avoiding predators and obstacles, and orienting in the sea. This sensory array provides information to a central nervous system (CNS) that includes a relatively large brain, particularly in the rays and galeomorph sharks, whose brain-to-body weight ratios are comparable to those of birds and mammals (Northcutt, 1978).

The plainfin midshipman fish,Porichthys notatus, is a seasonal breeding teleost fish for which vocal–acoustic communication is essential for its reproductive success. Female midshipman use the saccule as the primary end organ for hearing to detect and locate “singing” males that produce multiharmonic advertisement calls during the summer breeding season. Previous work has shown that female auditory sensitivity changes seasonally with reproductive state; summer reproductive females become better suited than winter nonreproductive females to detect and encode the dominant higher harmonic components in the male's advertisement call, which are potentially critical for mate selection and localization. Here, we test the hypothesis that these seasonal changes in female auditory sensitivity are concurrent with seasonal increases in saccular hair cell receptors. We show that there is increased hair cell density in reproductive females and that this increase is not dependent on body size since similar changes in hair cell density were not found in the other inner ear end organs. We also observed an increase in the number of small, potentially immature saccular hair bundles in reproductive females. The seasonal increase in saccular hair cell density and smaller hair bundles in reproductive females was paralleled by a dramatic increase in the magnitude of the evoked saccular potentials and a corresponding decrease in the auditory thresholds recorded from the saccule. This demonstration of correlated seasonal plasticity of hair cell addition and auditory sensitivity may in part facilitate the adaptive auditory plasticity of this species to enhance mate detection and localization during breeding.

A novel form of auditory plasticity for enhanced detection of social signals was described in a teleost fish, Porichthys notatus (Batrachoididae, Porichthyinae). The seasonal onset of male calling coincides with inshore migration from deep waters by both sexes and increased female sensitivity to dominant frequencies of male calls. The closely related Lusitanian toadfish, Halobatrachus didactylus, (Batrachoididae, Halophryninae) also breeds seasonally and relies on acoustic communication to find mates but, instead, both sexes stay in estuaries and show vocal activity throughout the year. We investigated whether the sensitivity of the inner ear saccule of H. didactylus is seasonally plastic and sexually dimorphic. We recorded evoked potentials from populations of saccular hair cells from non-reproductive and reproductive males and females in response to 15–945 Hz tones. Saccular hair cells were most sensitive at 15–205 Hz (thresholds between 111 and 118 dB re. 1 μPa). Both sexes showed identical hearing sensitivity and no differences were found across seasons. The saccule was well suited to detect conspecific vocalizations and low frequencies that overlapped with lateral line sensitivity. We showed that the saccule in H. didactylus has major importance in acoustic communication throughout the year and that significant sensory differences may exist between the two batrachoidid subfamilies.

The behavioral capacity for directional hearing and the discrimination of sound sources at different locations has been demonstrated in a number of fish species. Evidence suggests that all fishes can detect the acoustic particle motion component of sound using their ears. Few experiments, however, provide conclusive evidence that fishes can find the location of sound sources, and debate remains about whether fishes know where sound sources are in the same way as human listeners or if they have other strategies for finding sound sources. It is still uncertain how sound source localization by fishes is accomplished.