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Cross-modal plasticity occurs when the function of remaining senses is enhanced following deprivation or loss of a sensory modality. Auditory neural responses are enhanced in the auditory cortex, including increased sensitivity and frequency selectivity, following short-term visual deprivation in adult mice (Petrus et al. Neuron 81:664-673, 2014). Whether or not these visual deprivation-induced neural changes translate into improved auditory perception and performance remains unclear. As an initial investigation of the effects of adult visual deprivation on auditory behaviors, CBA/CaJ mice underwent binocular enucleation at 3-4 weeks old and were tested on a battery of learned behavioral tasks, acoustic startle response (ASR), and prepulse inhibition (PPI) tests beginning at least 2 weeks after the enucleation procedure. Auditory brain stem responses (ABRs) were also measured to screen for potential effects of visual deprivation on non-behavioral hearing function. Control and enucleated mice showed similar tone detection sensitivity and frequency discrimination in a conditioned lick suppression test. Both groups showed normal reactivity to sound as measured by ASR in a quiet background. However, when startle-eliciting stimuli were presented in noise, enucleated mice showed decreased ASR amplitude relative to controls. Control and enucleated mice displayed no significant differences in ASR habituation, PPI tests, or ABR thresholds, or wave morphology. Our findings suggest that while adult-onset visual deprivation induces cross-modal plasticity at the synaptic and circuit levels, it does not substantially influence simple auditory behavioral performance.

Numerous and non-acoustic experimental factors can potentially influence experimental outcomes in animal models when measuring the effects of noise exposures. Subject-related factors, including species, strain, age, sex, body weight, and post-exposure measurement timepoints, influence the observed hearing deficits. Experimenter effects, such as experience with experimental techniques and animal handling, may also factor into reported thresholds. In this study, the influence of subject sex, body mass, age at noise exposure, and timepoint of post-exposure recording are reported from a large sample of CBA/CaJ mice. Auditory brainstem response (ABR) thresholds differed between noise-exposed and unexposed mice, although the differences varied across tone frequencies. Thresholds across age at noise exposures and measurement delays after exposure also differed for some timepoints. Higher body mass correlated with higher ABR thresholds for unexposed male and female mice, but not for noise-exposed mice. Together, these factors may contribute to differences in phenotypic outcomes observed across studies or even within a single laboratory.

Deer mice (genus Peromyscus) are an emerging model for aging studies due to their longevity relative to rodents of similar size. Although Peromyscus species are well-represented in genetic, developmental, and behavioral studies, relatively few studies have investigated auditory sensitivity in this genus. Given the potential utility of Peromyscus for investigations of age-related changes to auditory function, we recorded auditory brainstem responses (ABRs) in two Peromyscus species, P. californicus, and P. leucopus, across the lifespan. We compared hearing sensitivity and ABR wave metrics measured in these species with measurements from Mus musculus (CBA/CaJ strain) to assess age-related effects on hearing across species. Recordings in young animals showed that all species had similar hearing ranges and thresholds with peak sensitivity ranging from 8 to 16 kHz; however, P. californicus and P. leucopus were more sensitive to frequencies below 8 kHz. Although M. musculus showed significant threshold shifts across a broad range of frequencies beginning at middle age and worsening among old individuals, older Peromyscus mice retained good sensitivity to sound across their lifespan. Middle-aged P. leucopus had comparable thresholds to young for frequencies below 24 kHz. P. leucopus also had notably large ABRs that were robust to age-related amplitude reductions, although response latencies increased with age. Old P. californicus were less sensitive to mid-range tones (8-16 kHz) than young individuals; however, there were no significant age-effects on ABR amplitudes or latencies in this species. These results indicate that longevity in Peromyscus mice may be correlated with delayed aging of the auditory system and highlight these species as promising candidates for longitudinal hearing research.

Age-related hearing loss is a very common sensory disability, affecting one in three older adults. Establishing a link between anatomical, physiological, and behavioral markers of presbycusis in a mouse model can improve the understanding of this disorder in humans. We measured age-related hearing loss for a variety of acoustic signals in quiet and noisy environments using an operant conditioning procedure and investigated the status of peripheral structures in CBA/CaJ mice. Mice showed the greatest degree of hearing loss in the last third of their lifespan, with higher thresholds in noisy than in quiet conditions. Changes in auditory brainstem response thresholds and waveform morphology preceded behavioral hearing loss onset. Loss of hair cells, auditory nerve fibers, and signs of stria vascularis degeneration were observed in old mice. The present work underscores the difficulty in ascribing the primary cause of age-related hearing loss to any particular type of cellular degeneration. Revealing these complex structure-function relationships is critical for establishing successful intervention strategies to restore hearing or prevent presbycusis.

The Johns Hopkins Otologic Research Laboratory was founded in 1924 as the first human temporal bone laboratory within the United States. To better understand the contributions of the Johns Hopkins Otologic Research Laboratory to our understanding of presbycusis, we consulted with a medical librarian and archivist to search the Alan Mason Chesney Medical Archives, PubMed, JSTOR, and Johns Hopkins Bulletin for published and unpublished works from the lab. Between 1924 and 1938, Samuel J. Crowe, the Chairman of Otolaryngology, and anatomist Stacy R. Guild amassed a collection of ∼1,800 temporal bones. This collection allowed for an unprecedented period of discovery related to otologic disease. They combined hearing thresholds measured by the recently invented audiometer with new techniques for temporal bone decalcification, sectioning, and staining, and a method for the graphic reconstruction of the cochlea. Crowe and Guild used this unique opportunity to correlate otopathology with hearing and to make the first detailed descriptions of the otopathology of presbycusis. In 1931 and 1934, they observed spiral ganglion neuron and outer hair cell loss in the basal turn of the cochlea in individuals with high-frequency hearing loss. These were the first studies to reveal that stria vascularis degeneration and middle ear pathology were not the most common causes for high-frequency hearing loss. Aside from revealing the primary driving factors of presbycusis, this work provided insight into the tonotopic organization of the cochlea. After initially being recruited to help raise money for the laboratory, medical illustrator Max Brödel used the vertical histologic cross-sections of the cochlea to produce illustrations of the ear. The decision to produce histologic sections in the plane of the superior semicircular canal likely influenced Brödel's illustrations that share a similar orientation and would later become widely circulated. Significant contributions from the Otologic Research Laboratory were also made by Mary Hardy, D.Sc., a woman who has previously received little recognition for her work. The sectioning of temporal bones was stopped in 1938 due to World War II, but much of Crowe's and Guild's work continued into the 1940s until a rift between the two resulted in the temporary closure of the laboratory in 1949. Nearly 100 years after its founding, discoveries from the Johns Hopkins Otologic Research Laboratory remain relevant and emphasize the importance of continued human temporal bone research to improve our understanding and treatment of otologic disease.

Echolocating bats rely on precise temporal processing of sound, as echolocation calls may be emitted at rates as high as 150–200 sounds per second in the terminal “buzz” phase that precedes prey capture by insectivorous species. High call repetition rates could introduce forward masking effects that interfere with echo detection; however, echolocating bats may have evolved auditory specializations to prevent repetition suppression of auditory responses and facilitate detection of sounds separated by very brief time intervals. We assessed the time course of post-stimulus recovery of the auditory brainstem response (ABR) in two bat species that differ in the temporal characteristics of their sonar behaviors: the insectivorous Eptesicus fuscus, which uses high sonar call rates to capture prey and the frugivorous Carollia perspicillata, which uses lower call rates to forage. We recorded forward-masking ABRs to paired clicks at varied inter-stimulus intervals and measured recovery from prior stimulation as the ratio of response amplitudes and latencies evoked by the second stimulus relative to the first. We observed significant species-specific effects of forward masking on ABR responses in which E. fuscus maintained comparable responses to ISIs less than 4 ms compared to longer post-stimulus response recovery times (>6 ms) in C. perspicillata.

The peripheral auditory system in normal-hearing individuals is highly tuned to discriminate very minor changes in frequency, but this frequency resolution may be susceptible to the effects of age and noise. Unfortunately, high-intensity noise damages the auditory periphery and leads to temporary or permanent hearing loss. Using a streamlined method to measure auditory brainstem responses (ABRs) in simultaneous spectrally notched noise, we measured masked thresholds and estimated peripheral frequency selectivity in male and female CBA/CaJ mice across the lifespan before and after noise exposure (8–16 kHz narrowband noise at 100 dBA SPL for 2 ). Mice were grouped by age at the time of noise exposure (44, 144, and 479 days old) and tracked longitudinally. We recorded ABRs to a 16 kHz tone in quiet, in the presence of a broadband masker, and in a masker with a spectral notch of varying widths (1/8th,1/4th, 1/2, 1, and 2 octaves centered around 16 kHz) before and after noise exposure for up to 6 months post-exposure. Our findings show that masking differs with age and time after noise exposure, and that young adult mice (

Funder: Calico Life Sciences, LLC, The naked mole‐rat (Heterocephalus glaber) has fascinated zoologists for at least half a century. It has also generated considerable biomedical interest not only because of its extraordinary longevity, but also because of unusual protective features (e.g. its tolerance of variable oxygen availability), which may be pertinent to several human disease states, including ischemia/reperfusion injury and neurodegeneration. A recent article entitled ‘Surprisingly long survival of premature conclusions about naked mole‐rat biology’ described 28 ‘myths’ which, those authors claimed, are a ‘perpetuation of beautiful, but falsified, hypotheses’ and impede our understanding of this enigmatic mammal. Here, we re‐examine each of these ‘myths’ based on evidence published in the scientific literature. Following Braude et al., we argue that these ‘myths’ fall into four main categories: (i) ‘myths’ that would be better described as oversimplifications, some of which persist solely in the popular press; (ii) ‘myths’ that are based on incomplete understanding, where more evidence is clearly needed; (iii) ‘myths’ where the accumulation of evidence over the years has led to a revision in interpretation, but where there is no significant disagreement among scientists currently working in the field; ( iv ) ‘myths’ where there is a genuine difference in opinion among active researchers, based on alternative interpretations of the available evidence. The term ‘myth’ is particularly inappropriate when applied to competing, evidence‐based hypotheses, which form part of the normal evolution of scientific knowledge. Here, we provide a comprehensive critical review of naked mole‐rat biology and attempt to clarify some of these misconceptions.

Age-related hearing loss (ARHL) is a public health problem that has been associated with negative health outcomes ranging from increased frailty to an elevated risk of developing dementia. Significant gaps remain in our knowledge of the underlying central neural mechanisms, especially those related to the efferent auditory pathways. Thus, the aim of this study was to quantify and compare age-related alterations in the cholinergic olivocochlear efferent auditory neurons. We assessed, in young-adult and aged CBA mice, the number of cholinergic olivocochlear neurons, auditory brainstem response (ABR) thresholds in silence and in presence of background noise, and the expression of excitatory and inhibitory proteins in the ventral nucleus of the trapezoid body (VNTB) and in the lateral superior olive (LSO). In association with aging, we found a significant decrease in the number of medial olivocochlear (MOC) cholinergic neurons together with changes in the ratio of excitatory and inhibitory proteins in the VNTB. Furthermore, in old mice we identified a correlation between the number of MOC neurons and ABR thresholds in the presence of background noise. In contrast, the alterations observed in the lateral olivocochlear (LOC) system were less significant. The decrease in the number of LOC cells associated with aging was 2.7-fold lower than in MOC and in the absence of changes in the expression of excitatory and inhibitory proteins in the LSO. These differences suggest that aging alters the medial and lateral olivocochlear efferent pathways in a differential manner and that the changes observed may account for some of the symptoms seen in ARHL.

The role of the mammalian auditory olivocochlear efferent system in hearing has long been the subject of debate. Its ability to protect against damaging noise exposure is clear, but whether or not this is the primary function of a system that evolved in the absence of industrial noise remains controversial. Here we review the behavioral consequences of olivocochlear activation and diminished olivocochlear function. Attempts to demonstrate a role for hearing in noise have yielded conflicting results in both animal and human studies. A role in selective attention to sounds in the presence of distractors, or attention to visual stimuli in the presence of competing auditory stimuli, has been established in animal models, but again behavioral studies in humans remain equivocal. Auditory processing deficits occur in models of congenital olivocochlear dysfunction, but these deficits likely reflect abnormal central auditory development rather than direct effects of olivocochlear feedback. Additional proposed roles in age-related hearing loss, tinnitus, hyperacusis, and binaural or spatial hearing, are intriguing, but require additional study. These behavioral studies almost exclusively focus on medial olivocochlear effects, and many relied on lesioning techniques that can have unspecific effects. The consequences of lateral olivocochlear and of corticofugal pathway activation for perception remain unknown. As new tools for targeted manipulation of olivocochlear neurons emerge, there is potential for a transformation of our understanding of the role of the olivocochlear system in behavior across species.

Aging leads to degeneration of the peripheral and central auditory systems, hearing loss, and difficulty understanding sounds in noise. Aging is also associated with changes in susceptibility to or recovery from damaging noise exposures, although the effects of the interaction between acute noise exposure and age on the perception of sounds are not well studied. We tested these effects in the CBA/CaJ mouse model of age-related hearing loss using operant conditioning procedures before and after noise exposure and longitudinally measured changes in their sensitivity for detecting tones in quiet or noise backgrounds. Cochleae from a subset of the behaviorally tested mice were immunolabeled to examine organ of Corti damage relative to what is expected based on aging alone. Mice tested in both quiet and noise background conditions experienced worse behavioral sensitivity immediately after noise exposure, but mice exposed at older ages generally showed greater threshold shifts and reduced recovery over time. Surprisingly, day-to-day stability in thresholds was markedly higher for mice detecting signals in the presence of a noise masker compared with detection in quiet conditions. Cochlear analysis revealed decreases in the total number of outer hair cells (OHCs) and the number of ribbons per inner cell in high-frequency regions in aged, noise-exposed mice relative to aging alone. Our findings build on previous work showing interactions between age and noise exposure and add that background noise can increase the stability of behavioral hearing sensitivity after noise damage.

Naked mole-rats are highly vocal, eusocial, subterranean rodents with, counterintuitively, poor hearing. The causes underlying their altered hearing are unknown. Moreover, whether altered hearing is degenerate or adaptive to their unique lifestyles is controversial. We used various methods to identify the factors contributing to altered hearing in naked and the related Damaraland mole-rats and to examine whether these alterations result from relaxed or adaptive selection. Remarkably, we found that cochlear amplification was absent from both species despite normal prestin function in outer hair cells isolated from naked mole-rats. Instead, loss of cochlear amplification appears to result from abnormal hair bundle morphologies observed in both species. By exploiting a well-curated deafness phenotype-genotype database, we identified amino acid substitutions consistent with abnormal hair bundle morphology and reduced hearing sensitivity. Amino acid substitutions were found in unique groups of six hair bundle link proteins. Molecular evolutionary analyses revealed shifts in selection pressure at both the gene and the codon level for five of these six hair bundle link proteins. Substitutions in three of these proteins are associated exclusively with altered hearing. Altogether, our findings identify the likely mechanism of altered hearing in African mole-rats, making them the only identified mammals naturally lacking cochlear amplification. Moreover, our findings suggest that altered hearing in African mole-rats is adaptive, perhaps tailoring hearing to eusocial and subterranean lifestyles. Finally, our work reveals multiple, unique evolutionary trajectories in African mole-rat hearing and establishes species members as naturally occurring disease models to investigate human hearing loss.

Objectives This study aims to determine if there is sex bias in basic and preclinical research on age-related hearing loss for the 10-year period of 2006–2015, prior to the NIH mandate of including sex as a biological variable in 2016. Design Manuscripts were identified in PubMed for the query “age-related hearing loss” for the 10-year period of 2006 to 2015. Manuscripts were included if they were original research (not reviews or meta-analyses), written in English, contained an abstract, used animals, and were primarily on age-related hearing loss. These criteria yielded 231 unique manuscripts for inclusion in the study analysis. The text of each manuscript was screened for the sex of the animals, the number of male and female animals, the discussion of sex-based results, the study site (US or international), and the year of publication. Results Only two thirds of manuscripts reported the sex of animals used in the experiments, and of these, 54% used both sexes, 34% used males only, and 13% used females only. In papers reporting sex and number of animals used, 67% were males and 33% were females. Over twice as many internationally based studies used males only compared to US-based studies. Only 15% of all manuscripts discussed sex-based results. Conclusions Sex bias is present in basic and preclinical age-related hearing loss research for the manuscripts screened in the 10-year period. Equal inclusion of both males and females in basic and preclinical age-related hearing loss research is critical for understanding sex-based differences in mechanisms and for effective treatment options.

Bats are long-lived animals that show presumed resistance to noise-induced and age-related hearing loss, which has been attributed to their dependence on sound processing for survival. Echolocation and basic auditory functions have been studied extensively in the big brown bat (Eptesicus fuscus), an insectivorous microchiropteran species. We conducted hearing tests and analysis of cochlear sensory cells in a group of big brown bats that exhibited anomalies in behavioral sonar tracking experiments and/or lacked neural responses to acoustic stimulation in subcortical auditory nuclei. We show for the first time the presence of profound deafness and extensive cochlear damage in an echolocating bat species. Auditory brainstem responses were abnormal or absent in these bats, and histological analyses of their cochleae revealed extensive loss of hair cells, supporting cells, and spiral ganglion neurons. The underlying cause of deafness is unknown.

CIB2 is a Ca(2+)- and Mg(2+)-binding protein essential for mechanoelectrical transduction (MET) by cochlear hair cells but not by vestibular hair cells that co-express CIB2 and CIB3. Here we show that in cochlear hair cells CIB3 can functionally substitute for CIB2. Using X-ray crystallography, we demonstrate that CIB2 and CIB3 are structurally similar to KChIP proteins, auxiliary subunits of voltage-gated K(v)4 channels. CIB2 and CIB3 bind to TMC1/2 through a domain in TMC1/2 flanked by transmembrane domains 2 and 3. The co-crystal structure of the CIB-binding domain in TMC1 with CIB3 reveals that interactions are mediated through a conserved CIB hydrophobic groove, similar to KChIP1 binding of K(v)4. Functional studies in mice show that CIB2 regulates TMC1/2 localization and function in hair cells, processes that are affected by deafness-causing CIB2 mutations. We conclude that CIB2 and CIB3 are MET channel auxiliary subunits with striking similarity to K(v)4 channel auxiliary subunits.

Acoustic startle response (ASR) modification procedures, especially prepulse inhibition (PPI), are increasingly used as behavioral measures of auditory processing and sensorimotor gating in rodents due to their perceived ease of implementation and short testing times. In practice, ASR and PPI procedures are extremely variable across animals, experimental setups, and studies, and the interpretation of results is subject to numerous caveats and confounding influences. We review considerations for modification of the ASR using acoustic stimuli, and we compare the sensitivity of PPI procedures to more traditional operant psychoacoustic techniques. We also discuss non-auditory variables that must be considered. We conclude that ASR and PPI measures cannot substitute for traditional operant techniques due to their low sensitivity. Additionally, a substantial amount of pilot testing must be performed to properly optimize an ASR modification experiment, negating any time benefit over operant conditioning. Nevertheless, there are some circumstances where ASR measures may be the only option for assessing auditory behavior, such as when testing mouse strains with early-onset hearing loss or learning impairments.

Introduction It is estimated over 466 million people worldwide have disabling hearing loss, and untreated hearing loss is associated with poorer health outcomes. The influence of sex as a biological variable on hearing loss is not well understood, especially for differences in underlying mechanisms which are typically elucidated through non-clinical research. Although the inclusion of sex as a biological variable in clinical studies has been required since 1993, sex reporting has only been recently mandated in National Institutes of Health funded non-clinical studies. Objective This article reviews the literature on recent non-clinical and clinical research concerning sex-based differences in hearing loss primarily since 1993, and discusses implications for knowledge gaps in the translation from non-clinical to clinical realms. Conclusions The disparity between sex-based requirements for non-clinical versus clinical research may inhibit a comprehensive understanding of sex-based mechanistic differences. Such disparities may play a role in understanding and explaining clinically significant sex differences and are likely necessary for developing robust clinical treatment options.

Cholinergic efferent neurons originating in the brainstem innervate the acoustico-lateralis organs (inner ear, lateral line) of vertebrates. These release acetylcholine (ACh) to inhibit hair cells through activation of calcium-dependent potassium channels. In the mammalian cochlea, ACh shunts and suppresses outer hair cell (OHC) electromotility, reducing the essential amplification of basilar membrane motion. Consequently, medial olivocochlear neurons that inhibit OHCs reduce the sensitivity and frequency selectivity of afferent neurons driven by cochlear vibration of inner hair cells (IHCs). The cholinergic synapse on hair cells involves an unusual ionotropic ACh receptor, and a near-membrane postsynaptic cistern. Lateral olivocochlear (LOC) neurons modulate type I afferents by still-to-be-defined synaptic mechanisms. Olivocochlear neurons can be activated by a reflex arc that includes the auditory nerve and projections from the cochlear nucleus. They are also subject to modulation by higher-order central auditory interneurons. Through its actions on cochlear hair cells, afferent neurons, and higher centers, the olivocochlear system protects against age-related and noise-induced hearing loss, improves signal coding in noise under certain conditions, modulates selective attention to sensory stimuli, and influences sound localization.

The effects of traumatic noise-exposure and deafening on auditory system function have received a great deal of attention. However, lower levels of noise as well as temporary conductive hearing loss also have consequences on auditory physiology and hearing. Here we review how abnormal acoustic experience at early ages affects the ascending and descending auditory pathways, as well as hearing behavior.

Individuals who have sustained a traumatic brain injury (TBI) often report auditory dysfunction including changes to their hearing sensitivity, tinnitus, and hyperacusis. One factor that may have a significant contribution to a person’s post-injury outcomes is whether the TBI occurred in combination with a high intensity noise exposure (e.g., as in the case of a car accident). To date, no study has examined the combined effects of a brain injury and high intensity noise exposure to evaluate whether this would lead to worse overall auditory outcomes than either trauma in isolation. In this study we tested the auditory sensitivity of male and female CBA/CaJ laboratory mice before and 3, 7, 14, 30, 60, and 90 days after acoustic (noise exposure), physical (controlled cortical impact), and combined acoustic and physical injuries. Following completion of the experiment, cochlear and brain tissue was fixed with paraformaldehyde and central and peripheral auditory structures were examined for damage. Our preliminary results show that physical injuries alone cause damage to the auditory sensitivity of mice measured using the auditory brainstem response, reduced acoustic startle responses in quiet and noise, yet hair cells remain intact. Long-term consequences on auditory perception are being tracked in these mice.

TMC1 and TMC2 (TMC1/2) have been proposed to form the pore of the mechanotransduction channel of cochlear hair cells. Here, we show that TMC1/2 cannot form mechanotransduction channels in cochlear hair cells without TMIE. TMIE binds to TMC1/2, and a TMIE mutation that perturbs TMC1/2 binding abolishes mechanotransduction. N-terminal TMIE deletions affect the response of the mechanotransduction channel to mechanical force. Similar to mechanically gated TREK channels, the C-terminal cytoplasmic TMIE domain contains charged amino acids that mediate binding to phospholipids, including PIP2. TMIE point mutations in the C terminus that are linked to deafness disrupt phospholipid binding, sensitize the channel to PIP2 depletion from hair cells, and alter the channel's unitary conductance and ion selectivity. We conclude that TMIE is a subunit of the cochlear mechanotransduction channel and that channel function is regulated by a phospholipid-sensing domain in TMIE with similarity to those in other mechanically gated ion channels.

Noise exposure is one of the most common causes of hearing loss and peripheral damage to the auditory system. A growing literature suggests that the auditory system can compensate for peripheral loss through increased central neural activity. The current study sought to investigate the link between noise exposure, increases in central gain, synaptic reorganization, and auditory function. All axons of the auditory nerve project to the cochlear nucleus, making it a requisite nucleus for sound detection. As the first synapse in the central auditory system, the cochlear nucleus is well positioned to respond plastically to loss of peripheral input. To investigate noise-induced compensation in the central auditory system, we measured auditory brainstem responses (ABRs) and auditory perception and collected tissue from mice exposed to broadband noise. Noise-exposed mice showed elevated ABR thresholds, reduced ABR wave 1 amplitudes, and spiral ganglion neuron loss. Despite peripheral damage, noise-exposed mice were hyperreactive to loud sounds and showed nearly normal behavioral sound detection thresholds. Ratios of late ABR peaks (2–4) relative to the first ABR peak indicated that brainstem pathways were hyperactive in noise-exposed mice, while anatomical analysis indicated there was an imbalance between expression of excitatory and inhibitory proteins in the ventral cochlear nucleus. The results of the current study suggest that a reorganization of excitation and inhibition in the ventral cochlear nucleus may drive hyperactivity in the central auditory system. This increase in central gain can compensate for peripheral loss to restore some aspects of auditory function.

Sound localization and directional hearing are fundamental for rodents to successfully navigate their environment, detect predators and prey, and locate conspecifics. In this chapter, the cues available to rodents for localizing and discriminating sounds in space are reviewed, including interaural level and timing differences and spectral cues. The neural circuits supporting sound localization are also introduced. Sound localization acuity and directional hearing behavior in azimuth and elevation that have been measured in terrestrial and subterranean rodent species are compared. Due to the limited scope of behavioral testing performed in most rodent species to date, binaural evoked potential studies that have been used to probe directional hearing in rodents are also considered as an alternative to labor-intensive behavioral measures. Finally, the trends observed across species are summarized and areas for future study are suggested.

Perceptual performance in persons with hearing loss, especially those using devices to restore hearing, is not fully predicted by traditional audiometric measurements designed to evaluate the status of peripheral function. The integrity of auditory brainstem synapses may vary with different forms of hearing loss, and differential effects on the auditory nerve-brain interface may have particularly profound consequences for the transfer of sound from ear to brain. Loss of auditory nerve synapses in ventral cochlear nucleus (VCN) has been reported after acoustic trauma, ablation of the organ of Corti, and administration of ototoxic compounds. The effects of gradually acquired forms deafness on these synapses are less well understood. We investigated VCN gross morphology and auditory nerve synapse integrity in DBA/2J mice with early-onset progressive sensorineural hearing loss. Hearing status was confirmed using auditory brainstem response audiometry and acoustic startle responses. We found no change in VCN volume, number of macroneurons, or number of VGLUT1-positive auditory nerve terminals between young adult and older, deaf DBA/2J. Cell-type specific analysis revealed no difference in the number of VGLUT1 puncta contacting bushy and multipolar cell body profiles, but the terminals were smaller in deaf DBA/2J mice. Transmission electron microscopy confirmed the presence of numerous healthy, vesicle-filled auditory nerve synapses in older, deaf DBA/2J mice. The present results suggest that synapses can be preserved over a relatively long time-course in gradually acquired deafness. Elucidating the mechanisms supporting survival of central auditory nerve synapses in models of acquired deafness may reveal new opportunities for therapeutic intervention.

Sound processing in the cochlea is modulated by cholinergic efferent axons arising from medial olivocochlear neurons in the brainstem. These axons contact outer hair cells in the mature cochlea and inner hair cells during development and activate nicotinic acetylcholine receptors composed of α9 and α10 subunits. The α9 subunit is necessary for mediating the effects of acetylcholine on hair cells as genetic deletion of the α9 subunit results in functional cholinergic de-efferentation of the cochlea. Cholinergic modulation of spontaneous cochlear activity before hearing onset is important for the maturation of central auditory circuits. In α9KO mice, the developmental refinement of inhibitory afferents to the lateral superior olive is disturbed, resulting in decreased tonotopic organization of this sound localization nucleus. In this study, we used behavioral tests to investigate whether the circuit anomalies in α9KO mice correlate with sound localization or sound frequency processing. Using a conditioned lick suppression task to measure sound localization, we found that three out of four α9KO mice showed impaired minimum audible angles. Using a prepulse inhibition of the acoustic startle response paradigm, we found that the ability of α9KO mice to detect sound frequency changes was impaired, whereas their ability to detect sound intensity changes was not. These results demonstrate that cholinergic, nicotinic α9 subunit mediate, transmission in the developing cochlear plays an important role in the maturation of hearing.

Studies have suggested a role of weakened medial olivocochlear (OC) efferent feedback in accelerated hearing loss and increased susceptibility to noise. The present study investigated the progression of hearing loss with age and exposure to a noisy environment in medial OC-deficient mice. Alpha9 nicotinic acetylcholine receptor knockout (α9KO) and wild types were screened for hearing loss using auditory brainstem responses. α9KO mice housed in a quiet environment did not show increased hearing loss compared to wild types in young adulthood and middle age. Challenging the medial OC system by housing in a noisy environment did not increase hearing loss in α9KO mice compared to wild types. ABR wave 1 amplitudes also did not show differences between α9KO mice and wild types. These data suggest that deficient medial OC feedback does not result in early onset of hearing loss.

Hair cells of the cochlea are mechanosensors for the perception of sound. Mutations in the LRTOMT gene, which encodes a protein with homology to the catecholamine methyltransferase COMT that is linked to schizophrenia, cause deafness. Here, we show that Tomt/Comt2, the murine ortholog of LRTOMT, has an unexpected function in the regulation of mechanotransduction by hair cells. The role of mTOMT in hair cells is independent of mTOMT methyltransferase function and mCOMT cannot substitute for mTOMT function. Instead, mTOMT binds to putative components of the mechanotransduction channel in hair cells and is essential for the transport of some of these components into the mechanically sensitive stereocilia of hair cells. Our studies thus suggest functional diversification between mCOMT and mTOMT, where mTOMT is critical for the assembly of the mechanotransduction machinery of hair cells. Defects in this process are likely mechanistically linked to deafness caused by mutations in LRTOMT/Tomt. DOI: http://dx.doi.org/10.7554/eLife.24318.001

Zellweger spectrum disorder (ZSD) is a disease continuum that results from inherited defects in PEX genes essential for normal peroxisome assembly. These autosomal recessive disorders impact brain development and also cause postnatal liver, adrenal, and kidney dysfunction, as well as loss of vision and hearing. The hypomorphic PEX1-G843D missense allele, observed in approximately 30% of ZSD patients, is associated with milder clinical and biochemical phenotypes, with some homozygous individuals surviving into early adulthood. Nonetheless, affected children with the PEX1-G843D allele have intellectual disability, failure to thrive, and significant sensory deficits. To enhance our ability to test candidate therapies that improve human PEX1-G843D function, we created the novel Pex1-G844D knock-in mouse model that represents the murine equivalent of the common human mutation. We show that Pex1-G844D homozygous mice recapitulate many classic features of mild ZSD cases, including growth retardation and fatty livers with cholestasis. In addition, electrophysiology, histology, and gene expression studies provide evidence that these animals develop a retinopathy similar to that observed in human patients, with evidence of cone photoreceptor cell death. Similar to skin fibroblasts obtained from ZSD patients with a PEX1-G843D allele, we demonstrate that murine cells homozygous for the Pex1-G844D allele respond to chaperone-like compounds, which normalizes peroxisomal β-oxidation. Thus, the Pex1-G844D mouse provides a powerful model system for testing candidate therapies that address the most common genetic cause of ZSD. In addition, this murine model will enhance studies focused on mechanisms of pathogenesis.

Exposure to high intensity sound can lead to temporary or permanent threshold shifts. Noise exposures that do not cause long-term hearing deficits, however, can induce extensive afferent ribbon synapse loss, while hair cells and spiral ganglion neurons remain mostly intact (Kujawa and Liberman, 2009). This loss of synapses despite normal hearing thresholds is referred to as hidden hearing loss (HHL). We examined the development of HHL in laboratory mice using operant conditioning with positive reinforcement. After exposing mice to 8–16 kHz narrowband noise at 100 dB SPL for 2 h, hearing thresholds temporarily shifted for both pure tone and ultrasonic vocalization stimuli; however, post-exposure thresholds and threshold shifts varied by sex and age. Immunohistochemistry and transmission electron microscopy were conducted to quantify peripheral damage and central synaptic reorganization once behavioral testing was complete. Brains were collected, sectioned, and labeled against VGLUT1 or GAD65 and labeling was quantified in the ventral cochlear nucleus. Cochleas were also collected, dissected, and labeled for myosin6 to label hair cells and either CTBP2 or SV2 to identify afferent or efferent terminals, respectively. Our findings show that mice are able to behaviorally recover hearing following non-traumatic noise exposure despite changes in peripheral and central auditory structures.

Accurate quantification of cell populations is essential in assessing and evaluating neural survival and degeneration in experimental groups. Estimates obtained through traditional two-dimensional counting methods are heavily biased by the counting parameters in relation to the size and shape of the neurons to be counted, resulting in a large range of inaccurate counts. In contrast, counting every cell in a population can be extremely labor-intensive. The present study hypothesizes that design-based stereology provides estimates of the total number of cochlear spiral ganglion neurons (SGNs) in mice that are comparable to those obtained by other accurate cell-counting methods, such as a serial reconstruction, while being a more efficient method. SGNs are indispensable for relaying auditory information from hair cells to the auditory brainstem, and investigating factors affecting their degeneration provides insight into the physiological basis for the progression of hearing dysfunction. Stereological quantification techniques offer the benefits of efficient sampling that is independent of the size and shape of the SGNs. Population estimates of SGNs in cochleae from young C57 mice with normal-hearing and C57 mice with age-related hearing loss were obtained using the optical fractionator probe and traditional two-dimensional counting methods. The average estimated population of SGNs in normal-hearing mice was 7009, whereas the average estimated population in mice with age-related hearing loss was 5096. The estimated population of SGNs in normal-hearing mice fell within the range of values previously reported in the literature. The reduction in the SGN population in animals with age-related hearing loss was statistically significant. Stereological measurements required less time per section compared to two-dimensional methods while optimizing the amount of cochlear tissue analyzed. These findings demonstrate that design-based stereology provides a practical alternative to other counting methods such as the Abercrombie correction method, which has been shown to notably underestimate cell populations, and labor-intensive protocols that account for every cell individually.

The notched noise method is an effective procedure for measuring frequency resolution and auditory filter shapes in both human and animal models of hearing. Briefly, auditory filter shape and bandwidth estimates are derived from masked thresholds for tones presented in noise containing widening spectral notches. As the spectral notch widens, increasingly less of the noise falls within the auditory filter and the tone becomes more detectible until the notch width exceeds the filter bandwidth. Behavioral procedures have been used for the derivation of notched noise auditory filter shapes in mice; however, the time and effort needed to train and test animals on these tasks renders a constraint on the widespread application of this testing method. As an alternative procedure, we combined relatively non-invasive auditory brainstem response (ABR) measurements and the notched noise method to estimate auditory filters in normal-hearing mice at center frequencies of 8, 11.2, and 16 kHz. A complete set of simultaneous masked thresholds for a particular tone frequency were obtained in about an hour. ABR-derived filter bandwidths broadened with increasing frequency, consistent with previous studies. The ABR notched noise procedure provides a fast alternative to estimating frequency selectivity in mice that is well-suited to high through-put or time-sensitive screening.

Increased prevalence of emotional distress is associated with tinnitus and hearing loss. The underlying mechanisms of the negative emotional response to tinnitus and hearing loss remain poorly understood, and it is challenging to disentangle the emotional consequences of hearing loss from those specific to tinnitus in listeners experiencing both. We addressed these questions in laboratory rats using three common rodent anxiety screening assays: elevated plus maze, open field test, and social interaction test. Open arm activity in the elevated plus maze decreased substantially after one trial in controls, indicating its limited utility for comparing pre- and post-treatment behavior. Open field exploration and social interaction behavior were consistent across multiple sessions in control animals. Individual sound-exposed and salicylate-treated rats showed a range of phenotypes in the open field, including reduced entries into the center in some subjects and reduced locomotion overall. In rats screened for tinnitus, less locomotion was associated with higher tinnitus scores. In salicylate-treated animals, locomotion was correlated with age. Sound-exposed and salicylate-treated rats also showed reduced social interaction. These results suggest that open field exploratory activity is a selective measure for identifying tinnitus distress in individual animals, whereas social interaction reflects the general effects of hearing loss. This animal model will facilitate future studies of the structural and functional changes in the brain pathways underlying emotional distress associated with hearing dysfunction, as well as development of novel interventions to ameliorate or prevent negative emotional responses.

Purpose: To assess possible damage to the hearing of experimental and companion animal subjects of magnetic resonance imaging (MRI) scans. Materials and Methods: Using animal hearing threshold data and sound level measurements from typical MRI pulse sequences, we estimated “equivalent loudness” experienced by several experimental and companion animals commonly subjects of MRI scans. We compared the equivalent loudness and exam duration to safe noise standards set by the National Institute for Occupational Safety and Health (NIOSH). Results: Monkeys, dogs, cats, pigs, and rabbits are frequently exposed to equivalent loudness levels during MRI scans beyond what is considered safe for human exposure. The sensitive frequency ranges for rats and mice are shifted substantially upward and their equivalent loudness levels fall within the NIOSH safe zone. Conclusion: MRI exposes many animals to levels of noise and duration that would exceed NIOSH human exposure limits. Researchers and veterinarians should use hearing protection for animals during MRI scans. Experimental research animals used in MRI studies are frequently kept and reimaged, and hearing loss could result in changed behavior. Damage to companion animals' hearing could make them less sensitive to commands and generally worsen interactions with family members. Much quieter MRI scanners would help decrease stress and potential harm to scanned animals, normalize physiology during MRI, and enable MRI of awake animals. J. Magn. Reson. Imaging 2012;36:743–747. © 2012 Wiley Periodicals, Inc.

Auditory neuropathy spectrum disorder (ANSD) describes a prevalent hearing disorder that is characterized by normal cochlear outer hair cell (OHC) function and abnormal auditory nerve function (1). Minimum diagnostic criteria include normal otoacoustic emissions (OAEs) and missing or aberrant auditory brainstem responses (ABRs). Changes in the ABR are assumed to arise from a dys-synchronization of sound-driven activity in the auditory nerve. The loss of temporal processing can impair speech, sound localization, and listening in noise. The impact of ANSD on speech has encouraged neonatal screening programs for early diagnosis of the disorder. These programs have revealed a higher incidence of ANSD among infants who have been treated in neonatal intensive care units (NICUs)(2,3). Although the health concerns that necessitate early hospitalization are varied, a constant factor in NICU treatments is the noise exposure that is generated by equipment, procedures, and staff (4,5). Several surveys have reported NICU noise levels consistently above the recommendations of the American Academy of Pediatrics (AAP) for both sustained (Leq) and maximum transient exposures (Lmax) (6). At present, significant ambiguities remain on what constitutes an acceptable level of noise exposure in human infants. While it is clear that a single loud exposure at a young age can accelerate hearing loss and promote spiral ganglion degeneration (7), the effects of persistent moderate exposures are difficult to estimate because environmental noise exerts pervasive effects on the developing auditory system. The deficits often manifest as perceptual disorders that are independent of overt hearing loss (8,9). Physiological studies of the cochlea and auditory nerve have provided detailed functional analyses of the medial olivocochlear system (MOCS). In normal individuals, the release of acetylcholine by the MOCS hyperpolarizes the membrane potentials of OHCs (10). The resulting decrease in cochlear mechanical sensitivity protects the ear from damaging levels of noise (11) and enhances auditory processing in distracting levels of noise (12,13). Recent clinical evidence suggests that ANSD is more prevalent among MOCS-compromised children (14). One explanation for this increased risk is that synaptic development of the central auditory pathways requires regulated sound-driven activity in the ascending inputs of the auditory nerve (15–17). When young individuals are exposed to environmental noise, MOC feedback may preserve normal synaptic development by regulating potentially disruptive levels of noise-driven activity. To investigate this novel developmental role for the MOCS, the feedback system was silenced in laboratory mice by a genetic modification of cholinergic signaling (18). The mice were reared in background noise, and then screened for ANSD using established diagnostic criteria. The audiological implications of the observed processing deficits were further delineated with a behavioral assay of temporal acuity. The refinement of this animal model allowed us to relate common clinical observations to direct electrophysiological recordings from the post-synaptic targets of the auditory nerve in the ventral cochlear nucleus. These recordings revealed significant functional impairments of the synaptic specializations that dictate the quality of temporal coding in the auditory brainstem.

Fast excitatory synaptic transmission in central synapses is mediated primarily by AMPA receptors (AMPARs), which are heteromeric assemblies of four subunits, GluA1-4. Among these subunits, rapidly gating GluA3/4 appears to be the most abundantly expressed to enable neurotransmission with submillisecond precision at fast rates in subsets of central synapses. However, neither definitive identification of the molecular substrate for native AMPARs in these neurons, nor their hypothesized functional roles in vivo has been unequivocally demonstrated, largely due to lack of specific antagonists. Using GluA3 or GluA4 knockout (KO) mice, we investigated these issues at the calyx of Held synapse, which is known as a high-fidelity synapse involved in sound localization. Patch-clamp recordings from postsynaptic neurons showed that deletion of GluA4 significantly slowed the time course of both evoked and miniature AMPAR-mediated excitatory postsynaptic currents (AMPAR-EPSCs), reduced their amplitude, and exacerbated AMPAR desensitization and short-term depression (STD). Surprisingly, presynaptic release probability was also elevated, contributing to severe STD at GluA4-KO synapses. In contrast, only marginal changes in AMPAR-EPSCs were found in GluA3-KO mice. Furthermore, independent of changes in intrinsic excitability of postsynaptic neurons, deletion of GluA4 markedly reduced synaptic drive and increased action potential failures during high-frequency activity, leading to profound deficits in specific components of the auditory brainstem responses associated with synchronized spiking in the calyx of Held synapse and other related neurons in vivo. These observations identify GluA4 as the main determinant for fast synaptic response, indispensable for driving high-fidelity neurotransmission and conveying precise temporal information.

The acoustic basis of auditory spatial acuity was investigated in CBA/129 mice by relating patterns of behavioral errors to directional features of the head-related transfer function (HRTF). Behavioral performance was assessed by training the mice to lick a water spout during sound presentations from a "safe" location and to suppress the response during presentations from "warning" locations. Minimum audible angles (MAAs) were determined by delivering the safe and warning sounds from different locations in the inter-aural horizontal and median vertical planes. HRTFs were measured at the same locations by implanting a miniature microphone and recording the gain of sound energy near the ear drum relative to free field. Mice produced an average MAA of 31° when sound sources were located in the horizontal plane. Acoustic measures indicated that binaural inter-aural level differences (ILDs) and monaural spectral features of the HRTF change systematically with horizontal location and therefore may have contributed to the accuracy of behavioral performance. Subsequent manipulations of the auditory stimuli and the directional properties of the ear produced errors that suggest the mice primarily relied on ILD cues when discriminating changes in azimuth. The MAA increased beyond 80° when the importance of ILD cues was minimized by testing in the median vertical plane. Although acoustic measures demonstrated a less robust effect of vertical location on spectral features of the HRTF, this poor performance provides further evidence for the insensitivity to spectral cues that was noted during behavioral testing in the horizontal plane.

Tinnitus is a pervasive auditory dysfunction affecting up to 10% of the adult population. The perception of ringing or hissing in the absence of a physical stimulus in one or both ears can be caused by acoustic trauma and other factors. Mice are a commonly used model for auditory disorders in humans, although the behavioral examination of tinnitus in mice has primarily been limited to reflexive measures involving inhibition of the acoustic startle by gaps in noise. Using an identification paradigm, we behaviorally tested whether mice show symptoms of tinnitus following long-term moderate noise exposure. Tinnitus was demonstrated by a shift in categorizing silence as narrowband noise. This experiment demonstrated that tinnitus can be induced in mice using noise exposure, similar to that caused by salicylate. Physiological and anatomical experiments reveal synaptic changes including response facilitation, increased reliability, increased synaptic terminal area, and increased number of release sites in mice ...

The Acoustical Society of America has a long, rich history of physiology research aimed at understanding the neural mechanisms underlying hearing and hearing dysfunction. In particular, this research has included pivotal studies in animal models that reveal the neurobiology of hearing and hearing disorders. More recently, research in animal models and research in human subjects have been considered in completely separate conferences or sessions. This situation presents a barrier to communicating important scientific information and fostering collaborations that bridge the gap between research in animal models and human listeners. It is crucial that the two types of research interface and inform one another to maximize translational potential and converge on fundamental knowledge. Experimental manipulations can be performed in non-human animals that cannot be performed in humans, but human and non-human animals can also be tested using similar techniques to increase translatability. Examples of mouse models tested using detailed analysis of synapse-level changes, clinically relevant evoked potentials, and behavioral tests will be presented. Areas for mutually beneficial interdisciplinary investigation will be discussed with the intention of inspiring new lines of communication between researchers focused on non-human animal and human subjects research.

Plasticity in the medial and lateral olivocochlear (MOC and LOC) systems in response to chronic exposure to background noise was investigated. Three different age-groups of young mice were exposed to the same chronic moderate noise conditions with either immediate tissue harvest or harvest following restoration to quiet conditions, with age matched controls. Cochleae were dissected and standard immunohistochemistry protocols were used to label hair cells with antibodies against myosin 6 and olivocochlear synaptic terminals with synaptic vesicle protein 2 (SV2). Specimens were imaged using confocal microscopy, and density of SV2 labeling was quantified. There was no statistically significant difference in MOC SV2 density between mice raised in noise and age matched controls for any group. However, there was a statistically significant increase in LOC SV2 density for adult mice raised in noise, particularly at higher frequency regions. This could suggest a protective upregulation of the efferent system against chronic moderate noise exposure. The increase in LOC innervation persisted for juvenile mice raised in noise with subsequent restoration to quiet conditions. These data suggest that the LOC system demonstrates sound-dependent plasticity, but that synaptic morphology may be altered for a substantial time period after exposure to noise ceases.