Auditory Research publications

We tested whether sensitivity to acoustic spectrotemporal modulations can be observed from reaction times for normal-hearing and impaired-hearing conditions. In a manual reaction-time task, normal-hearing listeners had to detect the onset of a ripple (with density between 0–8 cycles/octave and a fixed modulation depth of 50%), that moved up or down the log-frequency axis at constant velocity (between 0–64 Hz), in an otherwise-unmodulated broadband white-noise. Spectral and temporal modulations elicited band-pass filtered sensitivity characteristics, with fastest detection rates around 1 cycle/oct and 32 Hz for normal-hearing conditions. These results closely resemble data from other studies that typically used the modulation-depth threshold as a sensitivity criterion. To simulate hearing-impairment, stimuli were processed with a 6-channel cochlear-implant vocoder, and a hearing-aid simulation that introduced separate spectral smearing and low-pass filtering. Reaction times were always much slower compared to normal hearing, especially for the highest spectral densities. Binaural performance was predicted well by the benchmark race model of binaural independence, which models statistical facilitation of independent monaural channels. For the impaired-hearing simulations this implied a “best-of-both-worlds” principle in which the listeners relied on the hearing-aid ear to detect spectral modulations, and on the cochlear-implant ear for temporal-modulation detection. Although singular-value decomposition indicated that the joint spectrotemporal sensitivity matrix could be largely reconstructed from independent temporal and spectral sensitivity functions, in line with time-spectrum separability, a substantial inseparable spectral-temporal interaction was present in all hearing conditions. These results suggest that the reaction-time task yields a valid and effective objective measure of acoustic spectrotemporal-modulation sensitivity.

Significance: Quantitative measurements of cerebral hemodynamic changes due to functional activation are widely accomplished with commercial continuous wave (CW-NIRS) instruments despite the availability of the more rigorous multi-distance frequency domain (FD-NIRS) approach. A direct comparison of the two approaches to functional near-infrared spectroscopy can help in the interpretation of optical data and guide implementations of diffuse optical instruments for measuring functional activation. Aim: We explore the differences between CW-NIRS and multi-distance FD-NIRS by comparing measurements of functional activation in the human auditory cortex. Approach: Functional activation of the human auditory cortex was measured using a commercial frequency domain near-infrared spectroscopy instrument for 70 dB sound pressure level broadband noise and pure tone (1000 Hz) stimuli. Changes in tissue oxygenation were calculated using the modified Beer–Lambert law (CW-NIRS approach) and the photon diffusion equation (FD-NIRS approach). Results: Changes in oxygenated hemoglobin measured with the multi-distance FD-NIRS approach were about twice as large as those measured with the CW-NIRS approach. A finite-element simulation of the functional activation problem was performed to demonstrate that tissue oxygenation changes measured with the CW-NIRS approach is more accurate than that with multi-distance FD-NIRS. Conclusions: Multi-distance FD-NIRS approaches tend to overestimate functional activation effects, in part due to partial volume effects.

Stimulation of cholinergic efferent neurons innervating the inner ear has profound, well-characterized effects on vestibular and auditory physiology, after activating distinct ACh receptors (AChRs) on afferents and hair cells in peripheral endorgans. Efferent-mediated fast and slow excitation of vestibular afferents are mediated by α4β2*-containing nicotinic AChRs (nAChRs) and muscarinic AChRs (mAChRs), respectively. On the auditory side, efferent-mediated suppression of distortion product otoacoustic emissions (DPOAEs) is mediated by α9α10nAChRs. Previous characterization of these synaptic mechanisms utilized cholinergic drugs, that when systemically administered, also reach the CNS, which may limit their utility in probing efferent function without also considering central effects. Use of peripherally-acting cholinergic drugs with local application strategies may be useful, but this approach has remained relatively unexplored. Using multiple administration routes, we performed a combination of vestibular afferent and DPOAE recordings during efferent stimulation in mouse and turtle to determine whether charged mAChR or α9α10nAChR antagonists, with little CNS entry, can still engage efferent synaptic targets in the inner ear. The charged mAChR antagonists glycopyrrolate and methscopolamine blocked efferent-mediated slow excitation of mouse vestibular afferents following intraperitoneal, middle ear, or direct perilymphatic administration. Both mAChR antagonists were effective when delivered to the middle ear, contralateral to the side of afferent recordings, suggesting they gain vascular access after first entering the perilymphatic compartment. In contrast, charged α9α10nAChR antagonists blocked efferent-mediated suppression of DPOAEs only upon direct perilymphatic application, but failed to reach efferent synapses when systemically administered. These data show that efferent mechanisms are viable targets for further characterizing drug access in the inner ear.

The ability to perceive the world is not merely a passive process but depends on sensorimotor loops and interactions that guide and actively bias our sensory systems. Understanding which and how cognitive processes participate in this active sensing is still an open question. In this context, the auditory system presents itself as an attractive model for this purpose as it features an efferent control network that projects from the cortex to subcortical nuclei and even to the sensory epithelium itself. This efferent system can regulate the cochlear amplifier sensitivity through medial olivocochlear (MOC) neurons located in the brainstem. The ability to suppress irrelevant sounds during selective attention to visual stimuli is one of the functions that have been attributed to this system. MOC neurons are also directly activated by sounds through a brainstem reflex circuit, a response linked to the ability to suppress auditory stimuli during visual attention. Human studies have suggested that MOC neurons are also recruited by other cognitive functions, such as working memory and predictability. The aim of this research was to explore whether cognitive processes related to delayed responses in a visual discrimination task were associated with MOC function. In this behavioral condition, chinchillas held their responses for more than 2.5 s after visual stimulus offset, with and without auditory distractors, and the accuracy of these responses was correlated with the magnitude of the MOC reflex. We found that the animals’ performance decreased in presence of auditory distractors and that the results observed in MOC reflex could predict this performance. The individual MOC strength correlated with behavioral performance during delayed responses with auditory distractors, but not without them. These results in chinchillas, suggest that MOC neurons are also recruited by other cognitive functions, such as working memory.

Effective communication requires a match among signal characteristics, environmental conditions, and receptor tuning and decoding. The degree of matching, however, can vary, among others due to different selective pressures affecting the communication components. For evolutionary novelties, strong selective pressures are likely to act upon the signal and receptor to promote a tight match among them. We test this prediction by exploring the coupling between the acoustic signals and auditory sensitivity in Liolaemus chiliensis , the Weeping lizard, the only one of more than 285 Liolaemus species that vocalizes. Individuals emit distress calls that convey information of predation risk to conspecifics, which may respond with antipredator behaviors upon hearing calls. Specifically, we explored the match between spectral characteristics of the distress calls and the tympanic sensitivities of two populations separated by more than 700 km, for which previous data suggested variation in their distress calls. We found that populations differed in signal and receptor characteristics and that this signal variation was explained by population differences in body size. No precise match occurred between the communication components studied, and populations differed in the degree of such correspondence. We suggest that this difference in matching between populations relates to evolutionary processes affecting the Weeping lizard distress calls.

Current clinical strategies to assess benefits from hearing aids (HAs) are based on self-reported questionnaires and speech-in-noise (SIN) tests; which require behavioural cooperation. Instead, objective measures based on Auditory Brainstem Responses (ABRs) to speech stimuli would not require the individuals’ cooperation. Here, we re-analysed an existing dataset to predict behavioural measures with speech-ABRs using regression trees. Ninety-two HA users completed a self-reported questionnaire (SSQ-Speech) and performed two aided SIN tests: sentences in noise (BKB-SIN) and vowel-consonant-vowels (VCV) in noise. Speech-ABRs were evoked by a 40 ms [da] and recorded in 2x2 conditions: aided vs. unaided and quiet vs. background noise. For each recording condition, two sets of features were extracted: 1) amplitudes and latencies of speech-ABR peaks, 2) amplitudes and latencies of speech-ABR F0 encoding. Two regression trees were fitted for each of the three behavioural measures with either feature set and age, digit-span forward and backward, and pure tone average (PTA) as possible predictors. The PTA was the only predictor in the SSQ-Speech trees. In the BKB-SIN trees, performance was predicted by the aided latency of peak F in quiet for participants with PTAs between 43 and 61 dB HL. In the VCV trees, performance was predicted by the aided F0 encoding latency and the aided amplitude of peak VA in quiet for participants with PTAs ≤ 47 dB HL. These findings indicate that PTA was more informative than any speech-ABR measure, as these were relevant only for a subset of the participants. Therefore, speech-ABRs evoked by a 40 ms [da] are not a clinical predictor of behavioural measures in HA users.

Speech-on-speech informational masking arises because the interferer disrupts target processing (e.g., capacity limitations) or corrupts it (e.g., intrusions into the target percept); the latter should produce predictable errors. Listeners identified the consonant in monaural buzz-excited three-formant analogues of approximant-vowel syllables, forming a place of articulation series (/w/-/l/-/j/). There were two 11-member series; the vowel was either high-front or low-back. Series members shared formant-amplitude contours, fundamental frequency, and F1+F3 frequency contours; they were distinguished solely by the F2 frequency contour before the steady portion. Targets were always presented in the left ear. For each series, F2 frequency and amplitude contours were also used to generate interferers with altered source properties—sine-wave analogues of F2 (sine bleats) matched to their buzz-excited counterparts. Accompanying each series member with a fixed mismatched sine bleat in the contralateral ear produced systematic and predictable effects on category judgments; these effects were usually largest for bleats involving the fastest rate or greatest extent of frequency change. Judgments of isolated sine bleats using the three place labels were often unsystematic or arbitrary. These results indicate that informational masking by interferers involved corruption of target processing as a result of mandatory dichotic integration of F2 information, despite the grouping cues disfavoring this integration.

Big brown bats emit wideband frequency modulated (FM) ultrasonic pulses for echolocation. They perceive target range from echo delay and target size from echo amplitude. Their sounds contain two prominent down-sweeping harmonic sweeps (FM1, ∼55-22 kHz; FM2, ∼100-55 kHz), which are affected differently by propagation out to the target and back to the bat. FM2 is attenuated more than FM1 during propagation. Bats anchor target ranging asymmetrically on the low frequencies in FM1, while FM2 only contributes if FM1 is present as well. These experiments tested whether the bat’s ability to discriminate target size from the amplitude of echoes is affected by selectively attenuating upper or lower frequencies. Bats were trained to perform an echo amplitude discrimination task with virtual echo targets 83 cm away. While echo delay was held constant and echo amplitude was varied to estimate threshold, either lower FM1 frequencies or higher FM2 frequencies were attenuated. The results parallel effects seen in echo delay experiments; bats’ performance was significantly poorer when the lower frequencies in echoes were attenuated, compared to higher frequencies. The bat’s ability to distinguish between virtual targets at the same simulated range from echoes arriving at the same delay indicates a high level of focused attention for perceptual isolation of one and suppression of the other.

Hair cell mechanosensitivity resides in the sensory hair bundle, an apical protrusion of actin-filled stereocilia arranged in a staircase pattern. Hair bundle deflection activates mechano-electric transduction (MET) ion channels located near the tops of the shorter rows of stereocilia. The elicited macroscopic current is shaped by the hair bundle motion so that the mode of stimulation greatly influences the cell’s output. We present data quantifying the displacement of the whole outer hair cell bundle using high-speed imaging when stimulated with a fluid jet. We find a spatially non-uniform stimulation that results in splaying, where the hair bundle expands apart. Based on modeling, the splaying is predominantly due to fluid dynamics with a small contribution from hair bundle architecture. Additionally, in response to stimulation, the hair bundle exhibited a rapid motion followed by a slower motion in the same direction (creep) that is described by a double exponential process. The creep is consistent with originating from a linear passive system that can be modeled using two viscoelastic processes. These viscoelastic mechanisms are integral to describing the mechanics of the mammalian hair bundle.

The superior olivary complex (SOC) is a major computation center in the brainstem auditory system. Despite previous reports of high expression levels of cholinergic receptors in the SOC, few studies have addressed the functional role of acetylcholine in the region. The source of the cholinergic innervation is unknown for all but one of the nuclei of the SOC, limiting our understanding of cholinergic modulation. The medial nucleus of the trapezoid body, a key inhibitory link in monaural and binaural circuits, receives cholinergic input from other SOC nuclei and also from the pontomesencephalic tegmentum. Here, we investigate whether these same regions are sources of cholinergic input to other SOC nuclei. We also investigate whether individual cholinergic cells can send collateral projections bilaterally (i.e., into both SOCs), as has been shown at other levels of the subcortical auditory system. We injected retrograde tract tracers into the SOC in gerbils, then identified retrogradely-labeled cells that were also immunolabeled for choline acetyltransferase, a marker for cholinergic cells. We found that both the SOC and the pontomesencephalic tegmentum (PMT) send cholinergic projections into the SOC, and these projections appear to innervate all major SOC nuclei. We also observed a small cholinergic projection into the SOC from the lateral paragigantocellular nucleus of the reticular formation. These various sources likely serve different functions; e.g., the PMT has been associated with things such as arousal and sensory gating whereas the SOC may provide feedback more closely tuned to specific auditory stimuli. Further, individual cholinergic neurons in each of these regions can send branching projections into both SOCs. Such projections present an opportunity for cholinergic modulation to be coordinated across the auditory brainstem.

Ultrasound can deliver speech information when it is amplitude-modulated with speech and presented via bone conduction. This speech-modulated bone-conducted ultrasound (SM-BCU) can also transmit prosodic information. However, there is insufficient research on the recognition of vowel duration in SM-BCU. The aim of this study was to investigate the categorization of vowel durational changes in SM-BCU using a behavioral test. Eight Japanese-speaking participants with normal hearing participated in a forced-choice behavioral task to discriminate between “hato” (pigeon) and “haato” (heart). Speech signal stimuli were presented in seven duration grades from 220 ms to 340 ms. The threshold at which 50% of responses were “haato” was calculated and compared for air-conducted audible sound (ACAS) and SM-BCU. The boundary width was also evaluated. Although the SM-BCU threshold (mean: 274.6 ms) was significantly longer than the ACAS threshold (mean: 269.6 ms), there were no differences in boundary width. These results suggest that SM-BCU can deliver prosodic information about vowel duration with a similar difference limen to that of ACAS in normal hearing.

Hearing depends on glutamatergic synaptic transmission mediated by α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors (AMPARs). AMPARs are tetramers, where inclusion of the GluA2 subunit reduces overall channel conductance and Ca 2+ permeability. Cochlear afferent synapses between inner hair cells (IHCs) and auditory nerve fibers (ANFs) contain the AMPAR subunits GluA2, 3, and 4. However, the tetrameric complement of cochlear AMPAR subunits is not known. It was recently shown in mice that chronic intracochlear delivery of IEM-1460, an antagonist selective for GluA2-lacking AMPARs [also known as Ca 2+ -permeable AMPARs (CP-AMPARs)], before, during, and after acoustic overexposure prevented both the trauma to ANF synapses and the ensuing reduction of cochlear nerve activity in response to sound. Surprisingly, baseline measurements of cochlear function before exposure were unaffected by chronic intracochlear delivery of IEM-1460. This suggested that cochlear afferent synapses contain GluA2-lacking CP-AMPARs alongside GluA2-containing Ca 2+ -impermeable AMPA receptors (CI-AMPARs), and that the former can be antagonized for protection while the latter remain conductive. Here, we investigated hearing function in the guinea pig during acute local or systemic delivery of CP-AMPAR antagonists. Acute intracochlear delivery of IEM-1460 or systemic delivery of IEM-1460 or IEM-1925 reduced the amplitude of the ANF compound action potential (CAP) significantly, for all tone levels and frequencies, by > 50% without affecting CAP thresholds or distortion product otoacoustic emissions (DPOAE). Following systemic dosing, IEM-1460 levels in cochlear perilymph were ~ 30% of blood levels, on average, consistent with pharmacokinetic properties predicting permeation of the compounds into the brain and ear. Both compounds were metabolically stable with half-lives >5 h in vitro , and elimination half-lives in vivo of 118 min (IEM-1460) and 68 min (IEM-1925). Heart rate monitoring and off-target binding assays suggest an enhanced safety profile for IEM-1925 over IEM-1460. Compound potency on CAP reduction (IC 50 ~ 73 μM IEM-1460) was consistent with a mixture of GluA2-lacking and GluA2-containing AMPARs. These data strongly imply that cochlear afferent synapses of the guinea pig contain GluA2-lacking CP-AMPARs. We propose these CP-AMPARs may be acutely antagonized with systemic dosing, to protect from glutamate excitotoxicity, while transmission at GluA2-containing AMPARs persists to mediate hearing during the protection.

Many older adults have difficulty understanding speech in noisy backgrounds. In this study, we examined peripheral auditory, higher-level auditory, and cognitive factors that may contribute to such difficulties. A convenience sample of 137 volunteer older adults, 90 women, and 47 men, ranging in age from 47 to 94 years ( M = 69.2 and SD = 10.1 years) completed a large battery of tests. Auditory tests included measures of pure-tone threshold, clinical and psychophysical, as well as two measures of gap-detection threshold and four measures of temporal-order identification. The latter included two monaural and two dichotic listening conditions. In addition, cognition was assessed using the complete Wechsler Adult Intelligence Scale-3rd Edition (WAIS-III). Two monaural measures of speech-recognition threshold (SRT) in noise, the QuickSIN, and the WIN, were obtained from each ear at relatively high presentation levels of 93 or 103 dB SPL to minimize audibility concerns. Group data, both aggregate and by age decade, were evaluated initially to allow comparison to data in the literature. Next, following the application of principal-components factor analysis for data reduction, individual differences in speech-recognition-in-noise performance were examined using multiple-linear-regression analyses. Excellent fits were obtained, accounting for 60–77% of the total variance, with most accounted for by the audibility of the speech and noise stimuli and the severity of hearing loss with the balance primarily associated with cognitive function.

During vocal communication, the spectro-temporal structure of vocalizations conveys important contextual information. Bats excel in the use of sounds for echolocation by meticulous encoding of signals in the temporal domain. We therefore hypothesized that for social communication as well, bats would excel at detecting minute distortions in the spectro-temporal structure of calls. To test this hypothesis, we systematically introduced spectro-temporal distortion to communication calls of Phyllostomus discolor bats. We broke down each call into windows of the same length and randomized the phase spectrum inside each window. The overall degree of spectro-temporal distortion in communication calls increased with window length. Modelling the bat auditory periphery revealed that cochlear mechanisms allow discrimination of fast spectro-temporal envelopes. We evaluated model predictions with experimental psychophysical and neurophysiological data. We first assessed bats’ performance in discriminating original versions of calls from increasingly distorted versions of the same calls. We further examined cortical responses to determine additional specializations for call discrimination at the cortical level. Psychophysical and cortical responses concurred with model predictions, revealing discrimination thresholds in the range of 8–15 ms randomization-window length. Our data suggest that specialized cortical areas are not necessary to impart psychophysical resilience to temporal distortion in communication calls.

Neural hyperactivity induced by sound exposure often correlates with the development of hyperacusis and/or tinnitus. In laboratory animals, hyperactivity is typically induced by unilateral sound exposure to preserve one ear for further testing of hearing performance. Most ascending fibers in the auditory system cross into the superior olivary complex and then ascend contralaterally. Therefore, unilateral exposure should be expected to mostly affect the contralateral side above the auditory brain stem. On the other hand, it is well known that a significant number of neurons have crossing fibers at every level of the auditory pathway, which may spread the effect of unilateral exposure onto the ipsilateral side. Here we demonstrate that unilateral sound exposure causes development of hyperactivity in both the contra and ipsilateral inferior colliculus in mice. We found that both the spontaneous firing rate and bursting activity were increased significantly compared to unexposed mice. The neurons with characteristic frequencies at or above the center frequency of exposure showed the greatest increase. Surprisingly, this increase was more pronounced in the ipsilateral inferior colliculus. This study highlights the importance of considering both ipsi- and contralateral effects in future studies utilizing unilateral sound exposure.

The auditory biology of Monachinae seals is poorly understood. Limited audiometric data and certain anatomical features suggest that these seals may have reduced sensitivity to airborne sounds compared to related species. Here, we describe the in-air hearing abilities of a Hawaiian monk seal ( Neomonachus schauinslandi ) trained to participate in a psychophysical paradigm. We report absolute (unmasked) thresholds for narrowband signals measured in quiet conditions across the range of hearing and masked thresholds measured in the presence of octave-band noise at two frequencies. The behavioral audiogram indicates a functional hearing range from 0.1 to 33 kHz and poor sensitivity, with detection thresholds above 40 dB re 20 µPa. Critical ratio measurements are elevated compared to those of other seals. The apparently reduced terrestrial hearing ability of this individual—considered with available auditory data for a northern elephant seal ( Mirounga angustirostris )—suggests that hearing in Monachinae seals differs from that of the highly sensitive Phocinae seals. Exploration of phylogenetic relationships and anatomical traits support this claim. This work advances understanding of the evolution of hearing in amphibious marine mammals and provides updated information that can be used for management and conservation of endangered Hawaiian monk seals.

Three experiments explored the effects of abrupt changes in stimulus properties on streaming dynamics. Listeners monitored 20-s-long low- and high-frequency (LHL–) tone sequences and reported the number of streams heard throughout. Experiments 1 and 2 used pure tones and examined the effects of changing triplet base frequency and level, respectively. Abrupt changes in base frequency (±3–12 semitones) caused significant magnitude-related falls in segregation (resetting), regardless of transition direction, but an asymmetry occurred for changes in level (±12 dB). Rising-level transitions usually decreased segregation significantly, whereas falling-level transitions had little or no effect. Experiment 3 used pure tones (unmodulated) and narrowly spaced (±25 Hz) tone pairs (dyads); the two evoke similar excitation patterns, but dyads are strongly modulated with a distinctive timbre. Dyad-only sequences induced a strongly segregated percept, limiting scope for further build-up. Alternation between groups of pure tones and dyads produced large, asymmetric changes in streaming. Dyad-to-pure transitions caused substantial resetting, but pure-to-dyad transitions sometimes elicited even greater segregation than for the corresponding interval in dyad-only sequences (overshoot). The results indicate that abrupt changes in timbre can strongly affect the likelihood of stream segregation without introducing significant peripheral-channeling cues. These asymmetric effects of transition direction are reminiscent of subtractive adaptation in vision.

The effect of non-informational speech spectrum noise as a distractor on cognitive and listening comprehension ability was examined in fifty-three young, normal hearing adults. Time-controlled tasks were used to measure auditory working memory (WM) capacity and attention switching (AS) ability. Listening comprehension was measured using a lecture, interview, and spoken narratives test. Noise level was individually set to achieve at least 90% or higher speech intelligibility. Participants’ listening comprehension in the presence of distracting noise was better on inference questions compared to listening in quiet. Their speed of information processing was also significantly faster in WM and AS tasks in noise. These results were consistent with the view that noise may enhance arousal levels leading to faster information processing during cognitive tasks. Whereas the speed of AS was faster in noise, this rapid switching of attention resulted in more errors in updating items. Participants who processed information faster in noise and did so accurately, more effectively switched their attention to refresh/rehearse recall items within WM. More efficient processing deployed in the presence of noise appeared to have led to improvements in WM performance and making inferences in a listening comprehension task. Additional research is required to examine these findings using background noise that can cause informational masking.

Hair cells detect sound, head position or water movements when their mechanosensory hair bundle is deflected. Each hair bundle has an asymmetric architecture that restricts stimulus detection to a single axis. Coordinated hair cell orientations within sensory epithelia further tune stimulus detection at the organ level. Here, we identify GPR156, an orphan GPCR of unknown function, as a critical regulator of hair cell orientation. We demonstrate that the transcription factor EMX2 polarizes GPR156 distribution, enabling it to signal through Gαi and trigger a 180° reversal in hair cell orientation. GPR156-Gαi mediated reversal is essential to establish hair cells with mirror-image orientations in mouse otolith organs in the vestibular system and in zebrafish lateral line. Remarkably, GPR156-Gαi also instructs hair cell reversal in the auditory epithelium, despite a lack of mirror-image organization. Overall, our work demonstrates that conserved GPR156-Gαi signaling is integral to the framework that builds directional responses into mechanosensory epithelia.

Congenital hearing loss affects 1 in every 1000 births, with genetic mutations contributing to more than 50% of all cases. X-linked nonsyndromic hereditary hearing loss is associated with six loci (DFNX1-6) and five genes. Recently, the missense mutation (c.1771G>A, p.Gly591Ser) in COL4A6 , encoding the basement membrane (BM) collagen α6(IV) chain, was shown to be associated with X-linked congenital nonsyndromic hearing loss with cochlear malformation. However, the mechanism by which the COL4A6 mutation impacts hereditary hearing loss has not yet been elucidated. Herein, we investigated Col4a6 knockout (KO) effects on hearing function and cochlear formation in mice. Immunohistochemistry showed that the collagen α6(IV) chain was distributed throughout the mouse cochlea within subepithelial BMs underlying the interdental cells, inner sulcus cells, basilar membrane, outer sulcus cells, root cells, Reissner’s membrane, and perivascular BMs in the spiral limbus, spiral ligament, and stria vascularis. However, the click-evoked auditory brainstem response analysis did not show significant changes in the hearing threshold of Col4a6 KO mice compared with wild-type (WT) mice with the same genetic background. In addition, the cochlear structures of Col4a6 KO mice did not exhibit morphological alterations, according to the results of high-resolution micro-computed tomography and histology. Hence, loss of Col4a6 gene expression in mice showed normal click ABR thresholds and normal cochlear formation, which differs from humans with the COL4A6 missense mutation c.1771G>A, p.Gly591Ser. Therefore, the deleterious effects in the auditory system caused by the missense mutation in COL4A6 are likely due to the dominant-negative effects of the α6(IV) chain and/or α5α6α5(IV) heterotrimer with an aberrant structure that would not occur in cases with loss of gene expression.