Auditory Research publications

Noise exposure is one of the most common causes of sensorineural hearing loss. Although many studies considered inflammation to be a major contributor to noise-induced hearing loss, the process of cochlear inflammation is still unclear. Studies have found that activation of the NF-κB signaling pathway results in the accumulation of macrophages in the inner ear plays an important role in hair cell damage. In this study, tandem mass tag (TMT) technique was used to analyze the changes in basilar membrane proteome expression before and after acoustic injury. After noise exposure, the nicotinamide adenine dinucleotide (NAD) metabolism level was decreased, and the NF-κB signaling pathway was activated. The expression of CD38, the main NAD hydrolase in mammals, may directly lead to inflammation onset. Then, anakinra, an IL-1 receptor blocker, and apigenin, a CD38 inhibitor, were administered to animals to protect against noise-induced hearing loss. Our results showed that anakinra had little influence on the hearing threshold shift, while apigenin significantly reduce the threshold shift of hearing by inhibiting the expression of NF-κB and CD38 can be a promising target for protecting against noise-induced hearing loss.

Background Recent studies have identified hearing loss (HL) as a primary risk factor for Alzheimer’s disease (AD) onset. However, the mechanisms linking HL to AD are not fully understood. This study explored the effects of drug-induced hearing loss (DIHL) on the expression of proteins associated with AD progression in mouse models. Methods DIHL was induced in 5xFAD and Tg2576 mice aged 3 to 3.5 weeks using kanamycin (700 mg/kg, subcutaneous) and furosemide (600 mg/kg, intraperitoneal). The accumulation and expression of beta-amyloid (Aβ), ionized calcium-binding adaptor molecule 1 (Iba1), and glial fibrillary acidic protein (GFAP) were measured through immunohistochemistry and immunoblotting. Additionally, the expression of proteins involved in the mammalian target of rapamycin (mTOR) pathway, including downstream effectors p70 ribosomal S6 kinase (p70S6K) and S6, as well as proinflammatory cytokines, was analyzed. Results Compared to control conditions, HL led to a significant increase in the accumulation of Aβ in the hippocampus and cortex. Elevated levels of neuroinflammatory markers, including Iba1 and GFAP, as well as proinflammatory cytokines such as interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-alpha (TNF-α), were observed. Moreover, DIHL enhanced phosphorylation of mTOR, p70S6K, and S6, indicating activation of the mTOR pathway. Conclusions HL significantly increases Aβ accumulation in the brain. Furthermore, HL activates astrocytes and microglia, leading to increased neuroinflammation and thereby accelerating AD progression. These findings strongly suggest that HL contributes autonomously to neuroinflammation, highlighting the potential for early intervention in HL to reduce AD risk.

Objectives Cochlear implant array malpositioning is associated with impaired speech perception, vertigo, and facial nerve stimulation. Tip fold‐over is a subset of malpositioning that occurs more often with perimodiolar electrodes, but historically it has not been characterized due to lack of knowledge regarding electrode movements of the electrode within the cochlea. The aim of this study was to characterize the mechanics of tip fold‐over events and their associated insertion pressure profiles. Methods Cadaveric human heads were surgically prepared with a mastoidectomy and facial recess. Fiberoptic pressure sensors were inserted into the scala vestibuli and tympani to measure intracochlear pressures. Perimodiolar CI electrodes (Cochlear Slim‐Modiolar, CI532) were inserted via round window under fluoroscopy. Results Three types of tip fold‐over events were observed: anterior‐posterior C‐shaped, medial‐lateral C‐shaped, and S‐shaped roll‐overs. The largest transient pressures occurred with anterior‐posterior and S‐type roll‐over, and were associated with rotation or twisting inside the cochlea. Conclusions Results demonstrate at least three subtypes of tip fold‐overs. Elevated pressure transients were noted before and during the tip fold‐over event related to electrode twisting. The characterization of tip fold‐over into subtypes is novel and may aid identification of tip fold‐over events intraoperatively in the future. It remains important to identify tip fold‐over events, and they should be recognized early using a multimodal verification system. Further investigation is still required to determine the significance of these changes and other possible patterns of intracochlear electrode movement. Level of Evidence NA: Cadaver study Laryngoscope , 135:1795–1802, 2025

To encode continuous sound stimuli, the inner hair cell (IHC) ribbon synapses utilize calcium-binding proteins (CaBPs), which reduce the inactivation of their Ca V 1.3 calcium channels. Mutations in the CABP2 gene underlie non-syndromic autosomal recessive hearing loss DFNB93. Besides CaBP2, the structurally related CaBP1 is highly abundant in the IHCs. Here, we investigated how the two CaBPs cooperatively regulate IHC synaptic function. In Cabp1/2 double-knockout mice, we find strongly enhanced Ca V 1.3 inactivation, slowed recovery from inactivation and impaired sustained exocytosis. Already mild IHC activation further reduces the availability of channels to trigger synaptic transmission and may effectively silence synapses. Spontaneous and sound-evoked responses of spiral ganglion neurons in vivo are strikingly reduced and strongly depend on stimulation rates. Transgenic expression of CaBP2 leads to substantial recovery of IHC synaptic function and hearing sensitivity. We conclude that CaBP1 and 2 act together to suppress voltage- and calcium-dependent inactivation of IHC Ca V 1.3 channels in order to support sufficient rate of exocytosis and enable fast, temporally precise and indefatigable sound encoding.

Katydids employ acoustic signals to communicate with others of their species and have evolved to generate sounds by coupling the anatomical structures of their forewings. However, some species have evolved to implement an additional resonance mechanism that enhances the transmission and sound pressure of the acoustic signals produced by the primary resonators. Secondary resonators, such as burrow cavities or horn-shaped structures, are found in the surrounding environment but could also occur as anatomical modifications of their bodies. Chamber-like structures have been described in species of katydids with modified pronota or wings. It has been shown that these modified structures directly affect the transmission and filtering of acoustic signals and can function as a Helmholtz resonator that encapsulates the primary sound source. By morphological and acoustic analysis, we describe a new genus of Conocephalinae and investigate the physical properties of their sound production structures for three new species from the Andes of Colombia and Ecuador. Males of the new genus, here described as Tectucantus n. gen., have a characteristic inflated pronotum enclosing the reduced first pair of wings and extending rearward over the first abdominal segments. We test the hypothesis that the pronotal cavity volume correlates with the carrier frequency of specific calls. The cavity of the pronotal chamber acts as a Helmholtz resonator in all three Tectucantus species and, potentially, in other distantly related species, which use similar secondary body resonators.

Noise-induced hearing loss is a worldwide public health issue that is characterized by temporary or permanent changes in hearing sensitivity. This condition is closely linked to inflammatory responses, and interventions targeting the inflammatory gene tumor necrosis factor-alpha (TNFα) are known to mitigate cochlear noise damage. TNFα-induced proteins (TNFAIPs) are a family of translucent acidic proteins, and TNFAIP6 has a notable association with inflammatory responses. To date, there have been few reports on TNFAIP6 levels in the inner ear. To elucidate the precise mechanism, we generated transgenic mouse models with conditional knockout of Tnfaip6 (Tnfaip6 cKO). Evaluation of hair cell morphology and function revealed no significant differences in hair cell numbers or ribbon synapses between Tnfaip6 cKO and wild-type mice. Moreover, there were no notable variations in hair cell numbers or hearing function in noisy environments. Our results indicate that Tnfaip6 does not have a substantial impact on the auditory system.

The hispid pocket mouse ( C. hispidus ) is a solitary semi-fossorial rodent that has been the subject of various ecological and genetic studies. However, no previous studies have characterized its hearing ability, which is important for its survival and fitness. We collected auditory brainstem responses (ABRs) from C. hispidus and measured craniofacial and pinna morphological features to assess hearing ability and test differences in hearing thresholds, monaural and binaural ABR amplitudes and latencies between the sexes. ABR recordings revealed that similar to other small mammals, C. hispidus displayed the lowest threshold to sounds between 8-16 kHz, indicating best hearing across those frequencies. We found significant differences in auditory thresholds of the ABRs between the sexes, with females showing lower frequency hearing compared to males. However, no significant differences were detected in monaural and binaural ABR amplitudes and latencies between the sexes. We also found no significant differences in craniofacial and pinna dimensions between the sexes. These findings shed novel insights into the auditory systems across species and highlighted for the first time sex differences in auditory thresholds for this rodent species.

Bilateral cochlear implant (CI) patients exhibit significant limitations in spatial hearing. Their ability to process interaural time differences (ITDs) is often impaired, while their ability to process interaural level differences (ILDs) remains comparatively good. Clinical studies aiming to identify the causes of these limitations are often plagued by confounds and ethical limitations. Recent behavioral work suggests that rats may be a good animal model for studying binaural hearing under neuroprosthetic stimulation, as rats develop excellent ITD sensitivity when provided with suitable CI stimulation. However, their ability to use ILDs has not yet been characterized. Objective of this study is to address this knowledge gap. Neontally deafened rats were bilaterally fitted with CIs, and trained to lateralize binaural stimuli according to ILD. Their behavioral ILD thresholds were measured at pulse rates from 50 to 2400 pps. CI rats exhibited high sensitivity to ILDs with thresholds of a few dB at all tested pulse rates. We conclude that early deafened rats develop good sensitivity, not only to ITDs but also to ILDs, if provided with appropriate CI stimulation. Their generally good performance, in line with expectations from other mammalian species, validates rats as an excellent model for research on binaural auditory prostheses.

The mammalian auditory system encodes sounds with subtypes of spiral ganglion neurons (SGNs) that differ in sound level sensitivity, permitting discrimination across a wide range of levels. Recent work suggests the physiologically defined SGN subtypes correspond to at least three molecular subtypes. It is not known how information from the different subtypes converges within the cochlear nucleus. We examined this issue using transgenic mice of both sexes that express Cre recombinase in SGNs that are positive for markers of two subtypes: CALB2 (calretinin) in type 1a SGNs and LYPD1 in type 1c SGNs, which correspond to high- and low-sensitivity subtypes, respectively. We crossed these with mice expressing floxed channelrhodopsin, which allowed specific activation of axons from type 1a or 1c SGNs using optogenetics. We made voltage-clamp recordings from bushy cells in the anteroventral cochlear nucleus (AVCN) and found that the synapses formed by CALB2- and LYPD1-positive SGNs had similar EPSC amplitudes and short-term plasticity. Immunohistochemistry revealed that individual bushy cells receive a mix of 1a, 1b, and 1c synapses with VGluT1-positive puncta of similar sizes. We used optogenetic stimulation during in vivo recordings to classify chopper and primary-like units as receiving versus nonreceiving 1a- or 1c-type inputs. These groups showed no significant difference in threshold or spontaneous rate, suggesting the subtypes do not segregate into distinct processing streams in the AVCN. Our results indicate that principal cells in the AVCN integrate information from all SGN subtypes with extensive convergence, which could optimize sound encoding across a large dynamic range.

Exposure to loud noise is a common cause of acquired hearing loss. Disruption of subcellular calcium homeostasis and downstream stress pathways in the endoplasmic reticulum and mitochondria, including the unfolded protein response (UPR), have been implicated in the pathophysiology of noise-induced hearing loss. However, studies on the association between calcium homeostasis and stress pathways have been limited due to limited ability to measure calcium dynamics in mature-hearing, noise-exposed mice. We used a genetically encoded calcium indicator mouse model in which GCaMP6f is expressed specifically in hair cells or supporting cells under control of Myo15Cre or Sox2Cre, respectively. We performed live calcium imaging and UPR gene expression analysis in 8-week-old mice exposed to levels of noise that cause cochlear synaptopathy (98 db sound pressure level [SPL]) or permanent hearing loss (106 dB SPL). UPR activation occurred immediately after noise exposure, and the pattern of UPR activation was dependent on noise level, with the proapoptotic pathway upregulated only after 106 dB noise exposure. Spontaneous calcium transients in hair cells and intercellular calcium waves in supporting cells, which are present in neonatal cochleae, were quiescent in mature-hearing cochleae but reactivated upon noise exposure. Noise exposure of 106 dB was associated with more persistent and expansive intercellular Ca 2+ signaling wave activity. These findings demonstrate a strong and dose-dependent association between noise exposure, UPR activation, and changes in calcium homeostasis in hair cells and supporting cells, suggesting that targeting these pathways may be effective to develop treatments for noise-induced hearing loss.

In the mammalian cochlea, sensory hair cells are crucial for the transduction of acoustic stimuli into electrical signals, which are then relayed to the central auditory pathway via spiral ganglion neuron (SGN) afferent dendrites. The SGN output is directly modulated by inhibitory cholinergic axodendritic synapses from the efferent fibers originating in the superior olivary complex. When the adult cochlea is subjected to noxious stimuli or aging, the efferent system undergoes major rewiring, such that it reestablishes direct axosomatic contacts with the inner hair cells (IHCs), which occur only transiently during prehearing stages of development. The trigger, origin, and degree of efferent plasticity in the cochlea remains largely unknown. Using functional and morphological approaches, we demonstrate that efferent plasticity in the adult cochlea occurs as a direct consequence of mechanoelectrical transducer current dysfunction. We also show that, different from prehearing stages of development, the lateral olivocochlear — but not the medial olivocochlear — efferent fibers are those that form the axosomatic synapses with the IHCs. The study also demonstrates that in vivo restoration of IHC function using AAV- Myo7a rescue reestablishes the synaptic profile of adult IHCs and improves hearing, highlighting the potential of using gene-replacement therapy for progressive hearing loss.

The class C orphan G-protein-coupled receptor (GPCR) GPR156, which lacks the large extracellular region, plays a pivotal role in auditory function through Gi2/3. Here, we firstly demonstrate that GPR156 with high constitutive activity is essential for maintaining auditory function, and further reveal the structural basis of the sustained role of GPR156. We present the cryo-EM structures of human apo GPR156 and the GPR156-Gi3 complex, unveiling a small extracellular region formed by extracellular loop 2 (ECL2) and the N-terminus. The GPR156 dimer in both apo state and Gi3 protein-coupled state adopt a transmembrane (TM)5/6-TM5/6 interface, indicating the high constitutive activity of GPR156 in the apo state. Furthermore, C-terminus in G-bound subunit of GPR156 plays a dual role in promoting G protein binding within G-bound subunit while preventing the G-free subunit from binding to additional G protein. Together, these results explain how GPR156 constitutive activity is maintained through dimerization and provide a mechanistic insight into the sustained role of GPR156 in maintaining auditory function.

The brain predicts regularities in sensory inputs at multiple complexity levels, with neuronal mechanisms that remain elusive. Here, we monitored auditory cortex activity during the local-global paradigm, a protocol nesting different regularity levels in sound sequences. We observed that mice encode local predictions based on stimulus occurrence and stimulus transition probabilities, because auditory responses are boosted upon prediction violation. This boosting was due to both short-term adaptation and an adaptation-independent surprise mechanism resisting anesthesia. In parallel, and only in wakefulness, VIP interneurons responded to the omission of the locally expected sound repeat at the sequence ending, thus providing a chunking signal potentially useful for establishing global sequence structure. When this global structure was violated, by either shortening the sequence or ending it with a locally expected but globally unexpected sound transition, activity slightly increased in VIP and PV neurons, respectively. Hence, distinct cellular mechanisms predict different regularity levels in sound sequences.

Acoustic information in speech changes continuously, yet listeners form discrete perceptual categories to ease the demands of perception. Being a more continuous/gradient as opposed to a more discrete/categorical listener may be further advantageous for understanding speech in noise by increasing perceptual flexibility and resolving ambiguity. The degree to which a listener's responses to a continuum of speech sounds are categorical versus continuous can be quantified using visual analog scaling (VAS) during speech labeling tasks. Here, we recorded event-related brain potentials (ERPs) to vowels along an acoustic-phonetic continuum (/u/ to /a/) while listeners categorized phonemes in both clean and noise conditions. Behavior was assessed using standard two alternative forced choice (2AFC) and VAS paradigms to evaluate categorization under task structures that promote discrete vs. continuous hearing, respectively. Behaviorally, identification curves were steeper under 2AFC vs. VAS categorization but were relatively immune to noise, suggesting robust access to abstract, phonetic categories even under signal degradation. Behavioral slopes were correlated with listeners' QuickSIN scores; shallower slopes corresponded with better speech in noise performance, suggesting a perceptual advantage to noise degraded speech comprehension conferred by a more gradient listening strategy. At the neural level, P2 amplitudes and latencies of the ERPs were modulated by task and noise; VAS responses were larger and showed greater noise-related latency delays than 2AFC responses. More gradient responders had smaller shifts in ERP latency with noise, suggesting their neural encoding of speech was more resilient to noise degradation. Interestingly, source-resolved ERPs showed that more gradient listening was also correlated with stronger neural responses in left superior temporal gyrus. Our results demonstrate that listening strategy modulates the categorical organization of speech and behavioral success, with more continuous/gradient listening being advantageous to sentential speech in noise perception.

Because bats emit ultrasound and listen to echoes to perceive their surroundings, presenting phantom echoes is useful for understanding their perception of echolocation. Here, we present a protocol for the real-time playback of phantom echoes, especially from approaching objects. We describe steps for presenting virtual objects using a microphone, digital signal processor, and loudspeaker to induce the escape behavior of bats. We then detail procedures for clarifying their detection algorithm for approaching objects. For complete details on the use and execution of this protocol, please refer to Yoshida et al.1.

Inner ear hair cell (HC) damage is irreversible in mammals, but it has been shown that supporting cells (SCs) have the potential to differentiate into HCs. Serpine2, a serine protease inhibitor, encodes protease nexin 1, and this has been suggested to be a factor that promotes HC regeneration. In this study, we overexpressed Serpine2 in inner ear SCs cultured in two- and three-dimensional systems using the adeno-associated virus-inner ear (AAV-ie) vector, which promoted organoid expansion and HC differentiation. Overexpression of Serpine2 in the mouse cochlea through the round window membrane (RWM) injection promoted SC proliferation and HC regeneration, and the regenerated HCs were found to be derived from Lgr5+ SCs. Regenerated HCs have electrophysiological properties that are similar to those of native HCs. Notably, Serpine2 overexpression promoted HC survival and restored hearing of neomycin-damaged mice. In conclusion, our findings indicate that Serpine2 overexpression promotes HC regeneration and suggests that the utilization of inner ear progenitor cells in combination with AAVs might be a promising therapeutic target for hearing restoration.

Background: Speech sounds are processed in the human brain through intricate and interconnected cortical and subcortical structures. Two neural signatures, one largely from cortical sources (mismatch response, MMR) and one largely from subcortical sources (frequency-following response, FFR) are critical for assessing speech processing as they both show sensitivity to high-level linguistic information. However, there are distinct prerequisites for recording MMR and FFR, making them difficult to acquire simultaneously NEW METHOD: Using a new paradigm, our study aims to concurrently capture both signals and test them against the following criteria: (1) replicating the effect that the MMR to a native speech contrast significantly differs from the MMR to a nonnative speech contrast, and (2) demonstrating that FFRs to three speech sounds can be reliably differentiated. Results: Using EEG from 18 adults, we observed a decoding accuracy of 72.2 % between the MMR to native vs. nonnative speech contrasts. A significantly larger native MMR was shown in the expected time window. Similarly, a significant decoding accuracy of 79.6 % was found for FFR. A high stimulus-to-response cross-correlation with a 9 ms lag suggested that FFR closely tracks speech sounds. Comparison with existing method(s): These findings demonstrate that our paradigm reliably captures both MMR and FFR concurrently, replicating and extending past research with much fewer trials (MMR: 50 trials; FFR: 200 trials) and shorter experiment time (12 minutes). Conclusions: This study paves the way to understanding cortical-subcortical interactions for speech and language processing, with the ultimate goal of developing an assessment tool specific to early development.

Sound location coding has been extensively studied at the central nucleus of the mammalian inferior colliculus (CNIC), supporting a population code. However, this population code has not been extensively characterized on the single-trial level with simultaneous recordings or at other anatomical regions like the dorsal cortex of inferior colliculus (DCIC), which is relevant for learning-induced experience dependent plasticity. To address these knowledge gaps, here we made in two complementary ways large-scale recordings of DCIC populations from awake mice in response to sounds delivered from 13 different frontal horizontal locations (azimuths): volumetric two-photon calcium imaging with ~700 cells simultaneously recorded at a relatively low temporal resolution, and high-density single-unit extracellular recordings with ~20 cells simultaneously recorded at a high temporal resolution. Independent of the method, the recorded DCIC population responses revealed substantial trial-to-trial variation (neuronal noise) which was significantly correlated across pairs of neurons (noise correlations) in the passively listening condition. Nevertheless, decoding analysis supported that these noisy response patterns encode sound location on the single-trial basis, reaching errors that match the discrimination ability of mice. The detected noise correlations contributed to minimize the error of the DCIC population code of sound azimuth. Altogether these findings point out that DCIC can encode sound location in a similar format to what has been proposed for CNIC, opening exciting questions about how noise correlations could shape this code in the context of cortico-collicular input and experience-dependent plasticity.

Two adaptations of the Single-Interval Adjust-Matrix Yes-No (SIAM-YN) task, designed to increase the efficiency of absolute threshold estimation, are described. The first, the SIAM Twin Track (SIAM-TT) task, consists of two interleaved tracks of the standard SIAM-YN that are run in the same trial with a single response. The second new task modifies the binary SIAM-YN task by using a six-point rating-scale (SIAM-Rating). In Experiment 1, data from three tasks estimating absolute thresholds were obtained using a 10-ms tone, the 2-IFC up-down procedure, SIAM-YN task, and the SIAM-TT task. The data support the use of the SIAM-TT as an alternative to the conventional two-interval and one-interval (SIAM-YN) tasks when used to estimate absolute thresholds. By presenting two interleaved SIAM-YN tracks on a single experimental trial, the SIAM-TT task possesses greater efficiency alongside its signal-detection tradition which confers less response bias. Similarly, in Experiment 2, which compared the 2-IFC adaptive, SIAM-YN, and SIAM-Rating tasks, there was no main effect of task upon threshold estimates. The findings replicate previous studies supporting the validity and efficiency of the SIAM-YN task, and extends the SIAM-YN toolbox to efficiently facilitate the generation of psychometric functions (the SIAM-TT task) and Receiver Operating Characteristic Curves (the SIAM-Rating task).

Acoustic displays are conspicuous behaviors common across diverse animal taxa. They have long been studied in behavioral ecology, evolutionary biology, and neuroscience. Most of these investigations, however, have focused on male display. For species in which both sexes display, correcting this bias will lead to a fuller understanding of the evolution and function of these behaviors. In this study, we investigated the role of vocal advertisement display in female Alston’s singing mice ( Scotinomys teguina ). Singing mice are small muroid rodents of the family Cricetidae, native to high altitude cloud forests of Central America. Females and males both produce elaborate advertisement songs used in mate attraction and intra-sexual competition. Prior studies have largely focused on male behavior, though we recently found that both sexes dramatically increase their rate of singing and song duration in response to playback of conspecific male song. Here, we tested how mice of both sexes adjust their song effort in response to female song playback. Our results show that, like male song, female song elicits robust increases in song effort from both female and male mice. This study reveals additional social contexts that promote high song effort in this species, while raising additional questions about the role of song in communication within and between the sexes.