Publications using TDT

Age-related hearing loss (ARHL) is a disease that impacts human quality of life and contributes to the progression of other neuronal problems. Various stressors induce an increase in free radicals, destroy mitochondria to further contribute to cellular malfunction, and compromise cell viability, ultimately leading to functional decline. Cisd2, a master gene for Marfan syndrome, plays an essential role in maintaining mitochondrial integrity and functions. As shown by our data, specific deletion of Cisd2 in the cochlea exacerbated the hearing impairment of ARHL in C57BL/6 mice. Increased defects in mitochondrial function, potassium homeostasis and synapse activity were observed in the Cisd2-deleted mouse models. These mechanistic phenotypes combined with oxidative stress contribute to cell death in the whole cochlea. Human patients with obviously deteriorated ARHL had low Cisd2 expression; therefore, Cisd2 may be a potential target for designing therapeutic methods to attenuate the disease progression of ARHL.

In this paper we aim to examine the correlation between diffusion tensor imaging parameters of anatomical connectivity and characteristics of signal transmission obtained from patient-specific transfer function models. Here, we focused on elucidating the correlation between structural and functional neural connectivity within a cohort of pediatric patients diagnosed with dystonia. Diffusion tractography images were obtained from 12 patients with dystonia prior to the deep brain stimulation surgery. For each patient, we processed the imaging data to estimate anatomical measures including fractional anisotropy, axial diffusivity, number of fibre tracts per unit area, and fibre tract length. After the implantation of temporary depth leads for each patient as part of their treatment plan, intracranial signals were recorded. Transfer function models of local field potential recordings and the corresponding measures of functional connectivity were computed for each patient. Linear mixed effect analysis was then employed to determine the relationship between transfer function measures and diffusion tractography parameters. Our results illustrate a positive correlation between fractional anisotropy, AD, and intrinsic neural transmission measures, representing amplification and spread of intrinsic neural oscillations, obtained from the transfer functions models. However, no significant correlation was found between the functional connectivity and number of fibre tracts or fibre lengths. Our findings suggest that white matter integrity, as measured by fractional anisotropy and AD, can potentially reflect the amplification and spread of intrinsic brain signals throughout the network. This study underscores the significant relationship between structural and functional connectivity, offering valuable insights into propagation of neural activity in the brain network and potential implications for optimizing non-invasive treatments and planning for neurological disorders.

Objective This study aims to delineate the mechanisms through which intraperitoneal injection of gentamicin (GEN) influences the inner ear cells of mice by employing single-cell RNA sequencing (scRNA-seq) technology. Methods Eight-week-old Kunming mice were randomly assigned to three groups: a normal control group, a GEN group, and a GEN + dexamethasone (DEX) group. The mice received continuous intraperitoneal injections of the corresponding drugs for 10 days. Auditory brainstem response (ABR) was assessed to evaluate hearing threshold shifts, and cochlear tissues were harvested for scRNA-seq. The Seurat analysis workflow was employed for data quality control, dimensionality reduction clustering, and differential gene expression analysis. Results ABR results demonstrated a significant elevation in hearing thresholds in the GEN group, whereas thresholds in the DEX group showed improvement but remained elevated compared to the NOR group ( P < 0.05). Single-cell sequencing revealed notable alterations in the populations of outer hair cells, supporting cells, and immune cells in the GEN group. Analysis of differentially expressed genes identified significant downregulation of cell-specific genes Gbp6, Ppfia4 in hair cells of the GEN group, alongside upregulation of inflammation-related genes Nnat, Gh, indicating that hair cell damage and enhanced immune responses may be pivotal mechanisms underlying GEN-induced ototoxicity. Conclusion Utilizing scRNA-seq technology, this study uncovers substantial transcriptional changes induced by GEN in cochlear hair cells, supporting cells, and immune cells in mice, highlighting the role of inflammation and oxidative stress, TNF signaling pathways in its ototoxicity. DEX partially ameliorates hair cell damage.

The ability of the brain to process auditory information across multiple temporal scales is crucial for perception. This study introduces an innovative experimental approach using nonlinguistic structured stimuli composed of click trains that alternate at different rates to investigate auditory processing across 3 distinct temporal scales: individual clicks (tens of milliseconds), click trains forming auditory objects (hundreds of milliseconds), and higher-order click trains in neuronal novelty detection (seconds) in both rhesus monkeys and humans. Electrocorticography recordings in the auditory cortex of rhesus monkeys unveil the primate brain’s remarkable ability to process intricate auditory temporal patterns at timescales of tens and hundreds of milliseconds. Furthermore, extracellular recordings in monkeys demonstrate pronounced responsiveness to deviant click trains at longer temporal scales in the primary auditory cortex (A1), accompanied by synchronization with both individual clicks and click trains. By contrast, neurons in the auditory thalamus prefer individual clicks only. Notably, neurons in A1 exhibit the ability to synchronize with individual clicks while simultaneously integrating these clicks into cohesive train objects, a phenomenon we term “temporal integration during synchronization”. Additional human electroencephalography recordings complement and support these findings, highlighting the paradigm’s potential for clinical applications. Our research offers novel insights into the neural mechanisms underpinning auditory information processing across various temporal scales at the neuronal level.

This study investigates how selective auditory attention influences the lexical speech segmentation process to phonemes and words in a two competing speaker scenario. Using electroencephalography recordings from 20 participants, we applied temporal response function analysis to distinguish attention-driven neural activity to phoneme and word onsets for the attended and ignored speech stream separately. Our results reveal distinct attention effects for phoneme and word onsets. Phoneme onsets elicited significant selective attention effects at an early (18–94 ms, P1), middle (186–252 ms, P2), and late (302–382 ms, N2) time window. In contrast, word onsets showed attention effects only at a middle (192–280 ms, P2) and late (348–386 ms, N2) time window, occurring slightly later than phoneme-related effects. Prediction accuracy analyses demonstrated stronger model performance for the attended speech stream across all models, with notable improvements in prediction accuracy from a word model to a phoneme model to a combined word and phoneme model. These findings are in accordance with both hierarchical and parallel processing frameworks, where selective attention enhances lexical segmentation for attended speech, improving prediction accuracy. Early attention effects observed for phoneme onsets underscore their role in low-level speech processing, while late attention effects for word onsets may reflect higher level processing. This study highlights the importance of selective attention in neural speech tracking and provides insights into auditory processing mechanisms underlying speech comprehension in complex acoustic environments.

This study focuses on the critical role of HDAC11 in age-related hearing loss and its underlying mechanisms. Through cellular experiments, we deeply explored the effects of HDAC11 on the proliferation and senescence of HEI-OC1 cells. The results showed that HDAC11 overexpression significantly reduced the acetylation level of α-microtubule protein, which in turn affected the stability of microtubule structure and accelerated the apoptosis and senescence process of HEI-OC1 cells. In addition, the overexpression of HDAC11 inhibited the Pink1/Parkin signaling pathway, which impeded the mitochondrial autophagy process and ultimately led to mitochondrial dysfunction. In animal experiments, we further verified the ameliorative effect of HDAC11 overexpression on hearing loss in aged mice. The experimental results showed that HDAC11 overexpression not only attenuated the histopathological damage of the cochlea in aged mice but also effectively improved their hearing function. Notably, HDAC11 overexpression suppressed the expression of cellular autophagy-related proteins and Pink1 and Parkin proteins. In summary, the present study preliminarily revealed that HDAC11 may regulate mitochondrial autophagy by inhibiting the Pink1/Parkin pathway, thus providing a new theoretical basis for improving hearing loss in the elderly.

Objectives: The purinergic receptor P2X2 (P2RX2) encodes an ATP-gated ion channel permeable to Na+, K+, and especially Ca²⁺. Loss-of-function mutations in P2RX2 are known to cause autosomal dominant nonsyndromic deafness 41 (DFNA41), which manifests as high-frequency hearing loss, accelerated presbycusis, and increased susceptibility to noise-induced damage. Zebrafish, owing to their small size, rapid development, high fecundity, transparent embryos, and high gene conservation with humans, provide an ideal model for studying human diseases and developmental mechanisms. This study aims to generate a p2rx2 knockout zebrafish model using CRISPR/Cas9 gene editing system to investigate the effect of p2rx2 deficiency on the auditory system, providing a basis for understanding P2RX2-related hearing loss and developing gene therapy strategies. Methods: Two CRISPR targets (sgRNA1 and sgRNA2) spaced 47 bp apart were designed within the zebrafish p2rx2 gene. Synthesized sgRNAs and Cas9 protein were microinjected into single-cell stage Tübingen (TU)-strain zebrafish embryos. PCR and gel electrophoresis verified editing efficiency at 36 hours post-fertilization (hpf). Surviving embryos were raised to adulthood (F0), tail-clipped, genotyped, and screened for positive mosaics. F1 heterozygotes were generated by outcrossing, and F2 homozygous mutants were obtained by intercrossing. Polymerase chain reaction (PCR) combined with sequencing verified mutation type and heritability. At 5 days post-fertilization (dpf), YO-PRO-1 staining was used to examine hair cell morphology and count in lateral line neuromasts and the otolith region. Auditory evoked potential (AEP) thresholds at 600, 800, 1 000, and 2 000 Hz were measured in nine 4-month-old wild type and mutant zebrafish per group. Results: A stable p2rx2 knockout zebrafish line was successfully established. Sequencing revealed a 66 bp insertion at the first target site introducing a premature stop codon (TAA), leading to early termination of protein translation and loss of function. Embryos developed normally with no gross malformations. At 5 dpf, mutants exhibited significantly reduced hair cell density in the otolith region compared with wild type, although lateral line neuromasts were unaffected. AEP testing showed significantly elevated auditory thresholds at all 4 frequencies in homozygous mutants compared with wild type (all P<0.001), indicating reduced hearing sensitivity. Conclusions: We successfully generated a p2rx2 loss-of-function zebrafish model using CRISPR/Cas9 technology. p2rx2 deficiency caused hair cell defects in the otolith region and increased auditory thresholds across frequencies, indicating its key role in maintaining zebrafish auditory hair cell function and hearing perception. The phenotype's restriction to the otolith region suggests tissue-specific roles of p2rx2 in sensory organs. This model provides a valuable tool for elucidating the molecular mechanisms of P2RX2-related hearing loss and for screening otoprotective drugs and developing gene therapies.

Mutation in oxysterol-binding protein-like 2 ( OSBPL2) has been identified as the genetic cause of autosomal dominant nonsyndromic hearing loss (DFNA67, OMIM No. 616340). However, the pathogenesis of the OSBPL2 mutation in DFNA remains unclear. Our previous work showed that OSBPL2 deficiency impaired cell adhesion in auditory HEI-OC1 cells. In addition, loss of hair cells (HCs) and morphological abnormalities of HC stereocilia were detected in OSBPL2-disrupted pigs, suggesting that OSBPL2 plays an important role in the regulation of the actin cytoskeleton in auditory cells. In the present study, we found that OSBPL2 deficiency inhibited the Rho/ROCK2 signaling pathway and downregulated phosphorylated Ezrin-Radixin-Moesin (p-ERM), resulting in abnormal F-actin morphology in HEI-OC1 cells and stereociliary defects in mouse HCs. The present study demonstrates the underlying mechanism of OSBPL2 in the regulation of the actin cytoskeleton in HCs, which contributes to a deeper understanding of the pathogenesis of OSBPL2 mutations in DFNA.

Developing advanced tissue-engineered membranes with biocompatibility, suitable mechanical qualities, and anti-fibrotic and anti-inflammatory actions is important for tympanic membrane (TM) repair. Here, we present a novel acoustically transmitted decellularized fish swim bladder (DFB) loaded with mesenchymal stem cells (DFB@MSCs) for TM perforation (TMP) repair. The DFB scaffolds are obtained by removing the cellular components from the original FB, which retains the collagen composition that favors cell proliferation. Benefitting from their spatially porous structures and excellent mechanical properties, the DFB scaffolds can provide a suitable microenvironment and mechanical support for cell growth and tissue regeneration. In addition, by loading mesenchymal stem cells on the DFB scaffolds, the resultant DFB@MSCs system exhibits remarkable anti-fibrotic and anti-inflammatory effects, together with the ability to promote cell migration and angiogenesis. In vivo experiments confirm that the prepared DFB@MSCs scaffolds can not only alleviate inflammatory response caused by TMP but also promote new vessel formation, TM repair, and hearing improvement. These features indicate that our proposed DFB@MSCs stent is a prospective tool for the clinical repair of TM.

Despite the significant advantages of Shape Memory Polymers (SMPs), material processing and production challenges have limited their applications. Recent advances in fiber manufacturing offer a novel approach to processing polymers, broadening the functions of fibers beyond optical applications. In this study, a thermal drawing technique for SMPs to fabricate Shape Memory Polymer Fibers (SMPFs) tailored for medical applications, featuring programmable stiffness and shape control is developed. Rheological and differential scanning calorimetry analyses are conducted to assess SMP's compatibility with the proposed thermal drawing process and applications, leading to the production of multilumen, multimaterial SMPFs activated at body temperature. Different properties of SMPFs are investigated in three medical devices: stiffness‐adjustable catheters, softening neural interface, and shape‐programmable cochlear implants. Comprehensive characterization of these devices demonstrates the potential of thermally drawn SMPs to be employed in a wide range of applications demanding programmable mechanical properties.

Associative learning links sensory signals to their behavioral meaning by combining bottom-up inputs with top-down contextual information, enabling decision-making based on expected outcomes. It triggers plastic changes of neuronal responses in both excitatory and inhibitory cell populations. However, the role of inhibition in shaping this plasticity remains debated. Here we used chronic extracellular electrophysiology and optogenetic manipulation of inhibitory neurons and top-down inputs in mice learning an auditory discrimination task. We found that learning enhances stimulus selectivity in a subset of neurons in the primary auditory cortex. Interestingly, somatostatin-expressing (SST) interneurons decrease their response to the rewarded cue and bidirectionally regulate associative learning. More specifically, an increased activity of SST neurons impairs learning by altering bottom-up signaling, whereas the reduction of their activity facilitates learning by gating top-down inputs from the orbitofrontal cortex. These findings demonstrate that inhibition plays a critical role in gating top-down inputs to primary sensory cortices involved in associative learning.

Transitive inference allows people to infer new relations between previously experienced premises. It has been hypothesized that this logical thinking relies on a mental schema that spatially organizes elements, facilitating inferential insights. However, recent evidence challenges the need for these complex cognitive processes. To dig into the neural substrate driving TI cognitive processes, we examine the role of beta and gamma local field potential bands in the prefrontal cortex of 2 monkeys. During the inferential problem-solving period, we discover a tight link between beta and gamma bands modulation and TI complexity. This correlation diminishes its strength before initiating the motor response, indicating the chosen item. Notably, while the beta band maintains a constant relationship with TI performance throughout the trial, the gamma band shows a flexible relationship. This research highlights the role of beta and gamma interplay in cognitive computations when solving TI problems.

GJB2 encodes connexin 26 (Cx26), the most commonly mutated gene causing hereditary non‐syndromic hearing loss. Cx26 is mainly expressed in supporting cells (SCs) and fibrocytes in the mammalian cochlea. Gene therapy is currently considered the most promising strategy for eradicating genetic diseases. However, there have been no significant effects of gene therapy for GJB2 gene mutation‐associated deafness because deficiency of Cx26 leads to expanded sensory epithelial damage. In this study, the AAV2.7m8 serotype combined with the gfaABC1D promoter targeted infection of SCs is identified. It is found that Gjb2 gene replacement therapy in wild‐type mice results in sensory hair cells (HCs) deficits, excessive inflammatory responses, and hearing loss. This may be one of the key factors contributing to the hardship of GJB2 gene replacement therapy. Dexamethasone (DEX) shows promising results in inhibiting macrophage recruitment, with a protective effect against HC damage. Further, the combination of AAV2.7m8‐ Gjb2 with DEX shows a synergistic effect and enhances the gene therapy effect in a conditional Cx26 null mice model. These results indicate that the combination of gene therapy and medication will provide a new strategy for the treatment of hereditary deafness associated with GJB2 defects.

Ovarian-derived estrogen can signal non-canonically at membrane-associated receptors in the brain to rapidly regulate neuronal function. Early alcohol drinking confers greater risk for alcohol use disorder in women than men, and binge alcohol drinking is correlated with high estrogen levels, but a causal role for estrogen in driving alcohol drinking has not been established. We found that female mice displayed greater binge alcohol drinking and reduced avoidance when estrogen was high during the estrous cycle than when it was low. The pro-drinking, but not anxiolytic, effect of high endogenous estrogen occurred via rapid signaling at membrane-associated estrogen receptor alpha in the bed nucleus of the stria terminalis, which promoted synaptic excitation of corticotropin-releasing factor neurons and facilitated their activity during alcohol drinking. Thus, this study demonstrates a rapid, nongenomic signaling mechanism for ovarian-derived estrogen in the brain controlling behavior in gonadally intact females.

Deafness is the most common form of sensory impairment in humans and frequently caused by defects in hair cells of the inner ear. Here we demonstrate that in male mice which model recessive non-syndromic deafness (DFNB6), inactivation of Tmie in hair cells disrupts gene expression in the neurons that innervate them. This includes genes regulating axonal pathfinding and synaptogenesis, two processes that are disrupted in the inner ear of the mutant mice. Similar defects are observed in mouse models for deafness caused by mutations in other genes with primary functions in hair cells. Gene therapy targeting hair cells restores hearing and inner ear circuitry in DFNB6 model mice. We conclude that hair cell function is crucial for the establishment of peripheral auditory circuitry. Treatment modalities for deafness thus need to consider restoration of the function of both hair cells and neurons, even when the primary defect occurs in hair cells.

Alcohol use disorder (AUD) is a chronic relapsing brain disorder characterized by an impaired ability to stop or control alcohol consumption despite adverse social, occupational, or health consequences. AUD affects nearly one-third of adults at some point during their lives, with an associated cost of approximately $249 billion annually in the U.S. alone. The effects of alcohol consumption are expected to increase significantly during the COVID-19 pandemic, with alcohol sales increasing by approximately 54%, potentially exacerbating health concerns and risk-taking behaviors. Unfortunately, existing pharmacological and behavioral therapies for AUD are associated with poor success rates, with approximately 40% of individuals relapsing within three years of treatment.Pre-clinical studies have shown that chronic alcohol consumption leads to significant changes in synaptic function within the dorsal medial striatum (DMS), one of the brain regions associated with AUD and responsible for mediating goal-directed behavior. Specifically, chronic alcohol consumption has been associated with hyperactivity of dopamine receptor 1 (D1) medium spiny neurons (MSN) and hypoactivity of dopamine receptor 2 (D1) MSNs within the DMS. Optogenetic, chemogenetic, and transgenic approaches have demonstrated that reducing the D1/D2 MSN signaling imbalance decreases alcohol self-administration in rodent models of AUD.Here, we present an electrical stimulation approach that uses ultra-low (≤ 1 Hz) frequency (ULF) spike-timing-dependent plasticity (STDP) in mouse models of AUD to reduce DMS D1/D2 MSN signaling imbalances by stimulating D1-MSN afferents into the GPi and ACC glutamatergic projections to the DMS in a time-locked stimulation sequence. Our data suggest that GPi/ACC ULF-STDP selectively decreases DMS D1-MSN hyperactivity leading to reduced alcohol consumption without evoking undesired affective behaviors using electrical stimulation rather than approaches requiring genetic modification. This work represents a step towards fulfilling the unmet need for a reliable method of treating severe AUD through cell-type-specific control with clinically available neuromodulation tools.

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