ABSTRACT
The hair bundle of cochlear hair cells comprises specialized microvilli, the stereocilia, which fulfil the role of mechanotransduction. Genetic defects and environmental noise challenge the maintenance of hair bundle structure, critically contributing to age-related hearing loss. Stereocilia fusion is a major component of the hair bundle pathology in mature hair cells, but its role in hearing loss and its molecular basis are poorly understood. Here, we utilized super-resolution expansion microscopy to examine the molecular anatomy of outer hair cell stereocilia fusion in mouse models of age-related hearing loss, heightened endoplasmic reticulum stress and prolonged noise exposure. Prominent stereocilia fusion in our model of heightened endoplasmic reticulum stress, Manf (Mesencephalic astrocyte-derived neurotrophic factor)-inactivated mice in a background with Cadherin 23 missense mutation, impaired mechanotransduction and calcium balance in stereocilia. This was indicated by reduced FM1-43 dye uptake through the mechanotransduction channels, reduced neuroplastin/PMCA2 expression and increased expression of the calcium buffer oncomodulin inside stereocilia. Sparse BAIAP2L2 and myosin 7a expression was retained in the fused stereocilia but mislocalized away from their functional sites at the tips. These hair bundle abnormalities preceded cell soma degeneration, suggesting a sequela from stereociliary molecular perturbations to cell death signalling. In the age-related hearing loss and noise-exposure models, stereocilia fusion was more restricted within the bundles, yet both models exhibited oncomodulin upregulation at the fusion sites, implying perturbed calcium homeostasis. We conclude that stereocilia fusion is linked with the failure to maintain cellular proteostasis and with disturbances in stereociliary calcium balance. KEY POINTS: Stereocilia fusion is a hair cell pathology causing hearing loss. Inactivation of Manf, a component of the endoplasmic reticulum proteostasis machinery, has a cell-intrinsic mode of action in triggering outer hair cell stereocilia fusion and the death of these cells. The genetic background with Cadherin 23 missense mutation contributes to the high susceptibility of outer hair cells to stereocilia fusion, evidenced in Manf-inactivated mice and in the mouse models of early-onset hearing loss and noise exposure. Endoplasmic reticulum stress feeds to outer hair cell stereocilia bundle pathology and impairs the molecular anatomy of calcium regulation. The maintenance of the outer hair cell stereocilia bundle cohesion is challenged by intrinsic and extrinsic stressors, and understanding the underlying mechanisms will probably benefit the development of interventions to promote hearing health.
Subject(s)
Cadherins , Hair Cells, Auditory, Outer , Mechanotransduction, Cellular , Stereocilia , Animals , Stereocilia/metabolism , Stereocilia/pathology , Hair Cells, Auditory, Outer/metabolism , Hair Cells, Auditory, Outer/pathology , Mice , Cadherins/metabolism , Cadherins/genetics , Endoplasmic Reticulum Stress , Mice, Inbred C57BL , Male , Calcium/metabolism , Myosin VIIa/metabolism , Female , Hearing Loss/pathology , Hearing Loss/genetics , Hearing Loss/metabolism , Mutation, Missense , Calcium-Binding ProteinsABSTRACT
Cell cycle exit and acquirement of a postmitotic state is essential for the proper development of organs. In the present review, we examine the role of the cell cycle control in the sensory epithelia of the mammalian inner ear. We describe the roles of the core cell cycle regulators in the proliferation of prosensory cells and in the initiation and maintenance of terminal mitosis of the sensory epithelia. We also discuss how other intracellular signalling may influence the cell cycle. Finally, we address the question of whether manipulations of the cell cycle may have the potential to create replacement cells for the damaged inner sensory epithelia.
Subject(s)
Cell Cycle Proteins/genetics , Cell Cycle/genetics , Hair Cells, Auditory/physiology , Labyrinth Supporting Cells/physiology , Receptors, Notch/genetics , Animals , Cell Cycle Proteins/metabolism , Cell Differentiation , Cell Proliferation , Gene Expression Regulation, Developmental , Hair Cells, Auditory/cytology , Humans , Labyrinth Supporting Cells/cytology , Morphogenesis/physiology , Receptors, Notch/metabolism , Regeneration , Signal TransductionABSTRACT
Failure in the structural maintenance of the hair cell stereocilia bundle and ribbon synapse causes hearing loss. Here, we have studied how ER stress elicits hair cell pathology, using mouse models with inactivation of Manf (mesencephalic astrocyte-derived neurotrophic factor), encoding an ER-homeostasis-promoting protein. From hearing onset, Manf deficiency caused disarray of the outer hair cell stereocilia bundle and reduced cochlear sound amplification capability throughout the tonotopic axis. In high-frequency outer hair cells, the pathology ended in molecular changes in the stereocilia taper region and in strong stereocilia fusion. In high-frequency inner hair cells, Manf deficiency degraded ribbon synapses. The altered phenotype strongly depended on the mouse genetic background. Altogether, the failure in the ER homeostasis maintenance induced early-onset stereociliopathy and synaptopathy and accelerated the effect of genetic causes driving age-related hearing loss. Correspondingly, MANF mutation in a human patient induced severe sensorineural hearing loss from a young age onward. Thus, we present MANF as a novel protein and ER stress as a mechanism that regulate auditory hair cell maintenance in both mice and humans.
Subject(s)
Cochlea/metabolism , Hair Cells, Auditory, Inner/metabolism , Hair Cells, Auditory, Outer/metabolism , Nerve Growth Factors/genetics , Stereocilia/metabolism , Synapses/metabolism , Disease Susceptibility , Homeostasis , Nerve Growth Factors/metabolismABSTRACT
Sensory hair cells and supporting cells of the mammalian cochlea and vestibular (balance) organs exit the cell cycle during embryogenesis and do not proliferate thereafter. Here, we have studied the mechanisms underlying the maintenance of the postmitotic state and the proliferative capacity of these cells. We provide the first evidence of the role of cyclin D1 in cell cycle regulation in these cells. Cyclin D1 expression disappeared from embryonic hair cells as differentiation started. The expression was transiently upregulated in cochlear hair cells early postnatally, paralleling the spatiotemporal pattern of unscheduled cell cycle re-entry of cochlear hair cells from the p19(Ink4d)/p21(Cip1) compound mutant mice. Cyclin D1 misexpression in vitro in neonatal vestibular HCs from these mutant mice triggered S-phase re-entry. Thus, cyclin D1 suppression is important for hair cell's quiescence, together with the maintained expression of cyclin-dependent kinase inhibitors. In contrast to hair cells, cyclin D1 expression was maintained in supporting cells when differentiation started. The expression continued during the neonatal period when supporting cells have been shown to re-enter the cell cycle upon stimulation with exogenous mitogens. Thereafter, the steep decline in supporting cell's proliferative activity paralleled with cyclin D1 downregulation. Thus, cyclin D1 critically contributes to the proliferative plasticity of supporting cells. These data suggest that targeted cyclin D1 induction in supporting cells might be an avenue for proliferative regeneration in the inner ear.
Subject(s)
Cell Cycle , Cyclin D1/physiology , Cyclin-Dependent Kinase Inhibitor Proteins/physiology , Ear, Inner/embryology , Hair Cells, Auditory/cytology , Animals , Cell Proliferation , Cochlea/chemistry , Cyclin-Dependent Kinase Inhibitor p19/analysis , Cyclin-Dependent Kinase Inhibitor p19/physiology , Cyclin-Dependent Kinase Inhibitor p21/analysis , Cyclin-Dependent Kinase Inhibitor p21/physiology , Cyclin-Dependent Kinase Inhibitor p27/analysis , Cyclin-Dependent Kinase Inhibitor p27/physiology , Cyclin-Dependent Kinase Inhibitor p57/analysis , Cyclin-Dependent Kinase Inhibitor p57/physiology , Ki-67 Antigen/analysis , Mice , Signal Transduction , Wnt Proteins/physiology , beta Catenin/physiologyABSTRACT
Previous studies have shown that the maintenance of post-mitotic state is critical for the life-long survival of the inner ear mechanosensory cells, the hair cells. A general concept is that differentiated, post-mitotic cells rapidly die following cell cycle re-entry. Here we have compared the response of postnatal cochlear (auditory) and utricular (balance) hair cells to forced cell cycle reactivation and p53 up-regulation. Forced S-phase entry was triggered through the human papillomavirus-16 E7 oncogene misexpression in explant cultures. It induced DNA damage and p53 induction in cochlear outer hair cells and these cells were rapidly lost, before entry into mitosis. The death was attenuated by p53 inactivation. In contrast, despite DNA damage and p53 induction, utricular hair cells showed longer term survival and a proportion of them progressed into mitosis. Consistently, pharmacological elevation of p53 levels by nutlin-3a led to a death-prone phenotype of cochlear outer hair cells, while other hair cell populations were death-resistant. These data have important clinical implications as they show the importance of p53 in sensory cells that are essential in hearing function.
Subject(s)
Cell Cycle/physiology , Ear, Inner/cytology , Mechanoreceptors/physiology , Tumor Suppressor Protein p53/metabolism , Up-Regulation/physiology , Adenoviridae/physiology , Animals , Animals, Newborn , Calbindins , Cell Cycle/drug effects , Cell Differentiation/drug effects , Cell Differentiation/genetics , DNA Damage/physiology , Deoxyuridine/analogs & derivatives , Deoxyuridine/metabolism , Green Fluorescent Proteins/genetics , Imidazoles/pharmacology , Ki-67 Antigen/metabolism , Mice , Mice, Knockout , Organ Culture Techniques , Parvalbumins/genetics , Parvalbumins/metabolism , Piperazines/pharmacology , S100 Calcium Binding Protein G/genetics , S100 Calcium Binding Protein G/metabolism , Time Factors , Tumor Suppressor Protein p53/deficiency , Up-Regulation/drug effects , Up-Regulation/genetics , Viral Tropism/genetics , Viral Tropism/physiologyABSTRACT
INTRODUCTION: Tinnitus can become a strong stressor for some individuals, leading to imbalance of the autonomous nervous system with reduction of parasympathetic activity. It can manifest itself as sleep disturbances, anxiety and even depression. This condition can be reversed by bioelectrical vagal nerve stimulation (VNS). Conventional invasive VNS is an approved treatment for epilepsy and depression. Transcutaneous VNS (taVNS) stimulating the auricular branch of the vagus nerve has been shown to activate the vagal pathways similarly as an implanted VNS. Therefore, taVNS might also be a therapeutic alternative in health conditions such as tinnitus-related mental stress (TRMS). This retrospective study in 171 TRMS patients reports the clinical features, psychophysiological characteristics, and results of the heart rate variability (HRV) tests before and after test-taVNS. This study also reports the therapy outcomes of 113 TRMS patients treated with taVNS, in combination with standard tinnitus therapy. METHODS: Diagnostic tinnitus and hearing profiles were defined. To detect possible cardiac adverse effects, test-taVNS with heart rate monitoring as well as pre- and post-stimulation HRV tests were performed. Daily taVNS home therapy was prescribed thereafter. To assess therapeutic usefulness of taVNS, 1-year follow-up outcome was studied. Results of HRV tests were retrospectively analyzed and correlated to diagnostic data. RESULTS: The large majority of patients with TRMS suffer from associated symptoms such as sleep disturbances and anxiety. Baseline HRV data showed that more than three quarters of the 171 patients had increased sympathetic activity before test-taVNS. Test-taVNS shifted mean values of different HRV parameters toward increased parasympathetic activity in about 80% of patients. Test-taVNS did not cause any cardiac or other side effects. No significant adverse effects were reported in follow-up questionnaires. CONCLUSION: TRMS is an example of a stress condition in which patients may benefit from taVNS. As revealed by HRV, test-taVNS improved parasympathetic function, most efficiently in patients with a low starting HRV level. Our tinnitus treatment program, including taVNS, effectively alleviated tinnitus stress and handicap. For wider clinical use, there is a great need for more knowledge about the optimal methodology and parameters of taVNS.
ABSTRACT
The non-conventional neurotrophic factor mesencephalic astrocyte-derived neurotrophic factor (MANF) is an endoplasmic reticulum (ER)-resident protein that promotes ER homeostasis. MANF has a cytoprotective function, shown in the central nervous system neurons and pancreatic beta cells. Here, we report that MANF is expressed in the hair cells and neurons and in selected non-sensory cells of the cochlea and that Manf inactivation triggers upregulation of the ER chaperones in these cells. However, Manf inactivation resulted in the death of only outer hair cells (OHCs), the cells responsible for sound amplification in the cochlea. All OHCs were formed in Manf-inactivated mice, but progressive OHC death started soon after the onset of hearing function. The robust OHC loss was accompanied by strongly elevated hearing thresholds. Conditional Manf inactivation demonstrated that MANF has a local function in the cochlea. Immunostainings revealed the upregulation of CHOP, the pro-apoptotic component of the unfolded protein response (UPR), in Manf-inactivated OHCs, linking the UPR to the loss of these cells. The phenotype of Manf-inactivated OHCs was distinctly dependent on the mouse strain, such that the strains characterized by early-onset age-related hearing loss (C57BL/6J and CD-1) were affected. These results suggest that Manf deficiency becomes detrimental when accompanied by gene mutations that predispose to hearing loss, by intensifying ER dyshomeostasis. Together, MANF is the first growth factor shown to antagonize ER stress-mediated OHC death. MANF might serve as a therapeutic candidate for protection against hearing loss induced by the ER-machinery-targeting stressors.
Subject(s)
Cochlea/metabolism , Endoplasmic Reticulum Stress , Hair Cells, Auditory, Outer/metabolism , Hearing Loss/metabolism , Hearing , Nerve Growth Factors/deficiency , Animals , Auditory Threshold , Cell Death , Cochlea/pathology , Cochlea/physiopathology , Evoked Potentials, Auditory, Brain Stem , Hair Cells, Auditory, Outer/pathology , Hearing Loss/genetics , Hearing Loss/pathology , Hearing Loss/physiopathology , Mice, Inbred C57BL , Mice, Knockout , Nerve Growth Factors/genetics , Species SpecificityABSTRACT
Inner ear hair cells and supporting cells arise from common precursors and, in mammals, do not show phenotypic conversion. Here, we studied the role of the homeodomain transcription factor Prox1 in the inner ear sensory epithelia. Adenoviral-mediated Prox1 transduction into hair cells in explant cultures led to strong repression of Atoh1 and Gfi1, two transcription factors critical for hair cell differentiation and survival. Luciferase assays showed that Prox1 can repress transcriptional activity of Gfi1 independently of Atoh1. Prox1 transduction into cochlear outer hair cells resulted in degeneration of these cells, consistent with the known phenotype of Gfi1-deficient mice. These results together with the widespread expression of endogenous Prox1 within the population of inner ear supporting cells point to the role for Prox1 in antagonizing the hair cell phenotype in these non-sensory cells. Further, in vivo analyses of hair cells from Gfi1-deficient mice suggest that the cyclin-dependent kinase inhibitor p57(Kip2) mediates the differentiation- and survival-promoting functions of Gfi1. These data reveal novel gene interactions and show that these interactions regulate cellular differentiation within the inner ear sensory epithelia. The data point to the tight regulation of phenotypic characteristics of hair cells and supporting cells.
Subject(s)
Basic Helix-Loop-Helix Transcription Factors/metabolism , Cell Differentiation/physiology , DNA-Binding Proteins/metabolism , Ear, Inner/embryology , Epithelial Cells/physiology , Homeodomain Proteins/physiology , Transcription Factors/metabolism , Tumor Suppressor Proteins/physiology , Adenoviridae/genetics , Animals , Apoptosis/physiology , Cell Differentiation/genetics , Cells, Cultured , Cochlea/cytology , Cochlea/embryology , Cochlea/metabolism , Cyclin-Dependent Kinase Inhibitor p57/genetics , Cyclin-Dependent Kinase Inhibitor p57/metabolism , Ear, Inner/cytology , Epithelial Cells/cytology , Gene Expression Regulation, Developmental , Gene Transfer Techniques , Genes, Reporter , Hair Cells, Auditory/cytology , Hair Cells, Auditory/metabolism , Hair Cells, Auditory/virology , Homeodomain Proteins/genetics , Homeodomain Proteins/metabolism , Immunohistochemistry , Mice , Mice, Knockout , NIH 3T3 Cells , Organ Culture Techniques , Saccule and Utricle/cytology , Saccule and Utricle/embryology , Saccule and Utricle/metabolism , Tumor Suppressor Proteins/genetics , Tumor Suppressor Proteins/metabolismABSTRACT
CD2-associated protein (CD2AP) is an adapter protein that is involved in various signaling and vesicular trafficking processes and also functions as a linker between plasma membrane proteins and the actin cytoskeleton. The protein is known to have important functions in T cells and glomerular podocytes, but it is also expressed by many other adult-type tissues and cells. Here we analyzed the expression of the protein during early embryonic development and organogenesis of the mouse. The results showed differential tissue-specific regulation of CD2AP in developing and maturing organs. In oocytes and pre-implantation embryos, CD2AP was located diffusely in the cytoplasm, whereas in late blastocysts it was concentrated to the intercellular contacts. During organogenesis, CD2AP was distinctly upregulated upon, e.g., the pretubular aggregation of metanephric mesenchyme cells and the appearance of the osteoblastic rim around cartilages during endochondral ossification. High CD2AP expression was also observed during epithelial-like conversion of some highly specialized secretory cell types such as the odontoblasts, the cells of the choroid plexus and the decidualized cells of the endometrial stroma. In other instances, such as the development of the proximal tubuli of the kidney and the flat alveolar epithelium of the lung, the protein was downregulated upon differentiation and maturation of the cells. Finally, certain cells, e.g., glomerular podocytes, those forming the collecting ducts of the kidney, and the urothelium of the kidney pelvis, expressed CD2AP throughout their differentiation and maturation. Multiple molecules and complex pathways regulate embryogenesis, and scaffolding proteins apparently have pivotal roles in targeting and finetuning, e.g., growth factor- or hormone-induced processes. The cell-type specific spatio-temporal regulation of CD2AP during development suggests that this adapter protein is a key regulatory partner in many signaling pathways and cellular processes governing differentiation and morphogenesis.
Subject(s)
Adaptor Proteins, Signal Transducing/physiology , Cytoskeletal Proteins/physiology , Embryonic Development/physiology , Gene Expression Regulation, Developmental , Organogenesis/physiology , Adaptor Proteins, Signal Transducing/biosynthesis , Adaptor Proteins, Signal Transducing/genetics , Animals , Biological Transport/genetics , Biological Transport/physiology , Blastocyst/metabolism , Cell Differentiation , Cytoskeletal Proteins/biosynthesis , Cytoskeletal Proteins/genetics , Cytoskeleton/ultrastructure , Embryonic Development/genetics , Female , Germ Layers/metabolism , Gestational Age , Male , Mice , Oocytes/metabolism , Organ Specificity , Organogenesis/genetics , Ovary/metabolism , Placenta/metabolism , Pregnancy , Signal Transduction/genetics , Signal Transduction/physiology , Testis/metabolismABSTRACT
The mammalian auditory sensory epithelium, the organ of Corti, comprises the hair cells and supporting cells that are pivotal for hearing function. The origin and development of their precursors are poorly understood. Here we show that loss-of-function mutations in mouse fibroblast growth factor receptor 1 (Fgfr1) cause a dose-dependent disruption of the organ of Corti. Full inactivation of Fgfr1 in the inner ear epithelium by Foxg1-Cre-mediated deletion leads to an 85% reduction in the number of auditory hair cells. The primary cause appears to be reduced precursor cell proliferation in the early cochlear duct. Thus, during development, FGFR1 is required for the generation of the precursor pool, which gives rise to the auditory sensory epithelium. Our data also suggest that FGFR1 might have a distinct later role in intercellular signaling within the differentiating auditory sensory epithelium.
Subject(s)
Cell Communication/genetics , Cell Differentiation/genetics , Cell Division/genetics , Hair Cells, Auditory/abnormalities , Mutation/genetics , Receptor Protein-Tyrosine Kinases/deficiency , Receptors, Fibroblast Growth Factor/deficiency , Stem Cells/metabolism , Animals , Basic Helix-Loop-Helix Transcription Factors , Calbindins , Cell Death/genetics , DNA-Binding Proteins/genetics , Female , Fetus , Fibroblast Growth Factor 2/metabolism , Forkhead Transcription Factors , Gene Dosage , Gene Expression Regulation, Developmental/physiology , Hair Cells, Auditory/metabolism , Hair Cells, Auditory/ultrastructure , Integrases/genetics , Male , Mice , Mice, Mutant Strains , Nerve Tissue Proteins/genetics , Protein Isoforms/genetics , RNA, Messenger/metabolism , Receptor Protein-Tyrosine Kinases/genetics , Receptor, Fibroblast Growth Factor, Type 1 , Receptors, Fibroblast Growth Factor/genetics , S100 Calcium Binding Protein G/metabolism , Signal Transduction/genetics , Stem Cells/ultrastructure , Transcription Factors/metabolism , Viral Proteins/geneticsABSTRACT
Sensory hair cells of the auditory organ are generated during embryogenesis and remain postmitotic throughout life. Previous work has shown that inactivation of the cyclin-dependent kinase inhibitor (CKI) p19(Ink4d) leads to progressive hearing loss attributable to inappropriate DNA replication and subsequent apoptosis of hair cells. Here we show the synergistic action of another CKI, p21(Cip1), on cell cycle reactivation. The codeletion of p19(Ink4d) and p21(Cip1) triggered profuse S-phase entry of auditory hair cells during a restricted period in early postnatal life, leading to the transient appearance of supernumerary hair cells. In addition, we show that aberrant cell cycle reentry leads to activation of a DNA damage response pathway in these cells, followed by p53-mediated apoptosis. The majority of hair cells were absent in adult cochleas. These data, together with the demonstration of changing expression patterns of multiple CKIs in auditory hair cells during the stages of early postnatal maturation, show that the maintenance of the postmitotic state is an active, tissue-specific process, cooperatively regulated by several CKIs, and is critical for the lifelong survival of these sensory cells.
Subject(s)
Apoptosis/physiology , Cochlea/pathology , Cyclin-Dependent Kinase Inhibitor p19/physiology , Cyclin-Dependent Kinase Inhibitor p21/physiology , Genes, cdc , Hair Cells, Auditory/pathology , S Phase , Animals , Animals, Newborn , Animals, Suckling , Cell Count , Cochlea/growth & development , Crosses, Genetic , Cyclin-Dependent Kinase Inhibitor Proteins/biosynthesis , Cyclin-Dependent Kinase Inhibitor Proteins/genetics , Cyclin-Dependent Kinase Inhibitor p19/deficiency , Cyclin-Dependent Kinase Inhibitor p19/genetics , Cyclin-Dependent Kinase Inhibitor p21/deficiency , Cyclin-Dependent Kinase Inhibitor p21/genetics , DNA Damage , DNA Repair/genetics , Disease Progression , Hair Cells, Auditory/metabolism , HeLa Cells , Humans , Mice , Mice, Inbred C57BL , Mice, Knockout , Mitosis , Organ Specificity , Organ of Corti/pathology , Transfection , Tumor Suppressor Protein p53/physiologyABSTRACT
Various stressors, such as loud sounds and the effects of aging, impair the function and viability of the cochlear sensory cells, the hair cells. Stressors trigger pathophysiological changes in the cochlear non-sensory cells as well. We have here studied the stress response mounted in the lateral wall of the cochlea during acute noise stress and during age-related chronic stress. We have used the activation of JNK/c-Jun, ERK, and NF-κB pathways as a readout of the stress response, and the expression of the FoxO3 transcription factor as a possible additional player in cellular stress. In the aging cochlea, NF-κB transcriptional activity was strongly induced in the stria vascularis of the lateral wall. This induction was linked with the atrophy of the stria vascularis, suggesting a role for NF-κB signaling in mediating age-related strial degeneration. Acutely following noise exposure, the JNK/c-Jun, ERK, and NF-κB pathways were activated in the spiral ligament of the lateral wall of CBA/Ca mice. This activation was concomitant with the morphological transformation of macrophages, suggesting that the upregulation of stress signaling leads to macrophage activation. In contrast, C57BL/6J mice lacked these responses. Only the combination of noise exposure and a systemic stressor, lipopolysaccharide, exceeded the threshold for the activation of stress signaling in the lateral wall of C57BL/6J mice. In addition, we found that, at the young adult age, outer hair cells of CBA/Ca mice are much more vulnerable to loud sounds compared to these cells of C57BL/6J mice. These results suggest that the differential stress response in the lateral wall of the two mouse strains underlies, in part, the differential noise vulnerability of their outer hair cells. Together, we propose that the molecular stress response in the lateral wall modulates the outcome of the stressed cochlea.
Subject(s)
Aging/metabolism , Cochlea/metabolism , Noise/adverse effects , Proto-Oncogene Proteins c-jun/metabolism , Stress, Physiological , Animals , Cochlea/immunology , Cochlea/radiation effects , Forkhead Box Protein O3/metabolism , Lipopolysaccharides , MAP Kinase Signaling System , Mice, Inbred C57BL , Mice, Inbred CBA , Mice, Transgenic , NF-kappa B/metabolismABSTRACT
CONCLUSION: Music-induced acute acoustic trauma is not inevitably linked to hearing dysfunction as validated by conventional pure tone audiometry. Tinnitus is often in combination with hyperacusis. Our results point at 'silent hearing loss' as the underlying pathology, having afferent nerve terminal damage rather than hair cell loss as the structural correlate. OBJECTIVES: Exposure to loud music is one of the most common causes of acute acoustic trauma, which adolescents and teenagers experience by voluntary exposure to loud music of sound levels up to 110 dB(A). METHODS: The clinical and psychophysical data of 104 consecutive patients with music-induced hearing disorder (MIHD) were analyzed to construct individual hearing and tinnitus profiles. In all cases, tinnitus was the presenting symptom. RESULTS: Hearing abilities were normal in about two-thirds of the tinnitus patients. Tinnitus was experienced most often as a high-frequency tone (83%). The Tinnitus Handicap Inventory (THI) scores ranged from 0 to 94 with an average score of 43.1. Visual analog scales (VAS) were used to assess tinnitus loudness (average 42.4) and annoyance (average 54.2), and tinnitus awareness was estimated (average 60.3). All VAS values correlated strongly with the THI. Hyperacusis was present in 65% and 71% of the patients reported sleeping disorders.
Subject(s)
Music , Tinnitus/etiology , Adolescent , Adult , Audiometry, Pure-Tone , Auditory Threshold , Female , Hearing Loss, Noise-Induced/diagnosis , Hearing Loss, Noise-Induced/etiology , Humans , Hyperacusis/etiology , Male , Middle Aged , Surveys and Questionnaires , Tinnitus/diagnosis , Young AdultABSTRACT
CONCLUSION: In occupational noise-induced hearing loss (NIHL) reports, many tinnitus sufferers probably remain undetected and untreated at present. Attention should be focused on tinnitus, as well as threshold shifts resulting from NIHL. OBJECTIVES: Occupational NIHL is frequent among workers in industrialized countries and it is one of the greatest occupational health hazards. Hearing conservation programs have led to a reduction in the numbers of severe occupational NIHL. Our objectives were to analyze the severity of occupational NIHL reported in Finland, identify risk occupations, and investigate the occurrence of tinnitus among the reported cases. MATERIALS AND METHODS: We examined the records of 857 NIHL cases with an identifiable disability category of the total 858 NIHL cases reported in 2000. We sent tinnitus questionnaires to 366 of these NIHL cases. RESULTS: The degree of speech-frequency hearing loss was generally low, and a mention of tinnitus was reported in only 34 cases (4.0%). However, 88.7% of the patients actually had unreported tinnitus.
Subject(s)
Hearing Loss, Noise-Induced/epidemiology , Occupational Diseases/epidemiology , Tinnitus/epidemiology , Adolescent , Adult , Aged , Aged, 80 and over , Disability Evaluation , Female , Finland/epidemiology , Humans , Male , Middle Aged , Occupational Exposure/adverse effects , Occupations/statistics & numerical data , Registries , Surveys and QuestionnairesABSTRACT
CONCLUSION: Transcutaneous vagal nerve stimulation (tVNS) might offer a targeted, patient-friendly, and low-cost therapeutic tool for tinnitus patients with sympathovagal imbalance. OBJECTIVES: Conventionally, VNS has been performed to treat severe epilepsy and depression with an electrode implanted to the cervical trunk of vagus nerve. This study investigated the acute effects of tVNS on autonomic nervous system (ANS) imbalance, which often occurs in patients with tinnitus-triggered stress. METHODS: This study retrospectively analysed records of 97 patients who had undergone ANS function testing by heart rate variability (HRV) measurement immediately before and after a 15-60 min tVNS stimulation. RESULTS: The pre-treatment HRV recording showed sympathetic preponderance/reduced parasympathetic activity in about three quarters (73%) of patients. Active tVNS significantly increased variability of R-R intervals in 75% of patients and HRV age was decreased in 70% of patients. Either the variability of R-R intervals was increased or the HRV age decreased in 90% of the patients. These results indicate that tVNS can induce a shift in ANS function from sympathetic preponderance towards parasympathetic predominance. tVNS caused no major morbidity, and heart rate monitoring during the tVNS treatment showed no cardiac or circulatory effects (e.g. bradycardia) in any of the patients.
Subject(s)
Sympathetic Nervous System/physiopathology , Tinnitus/therapy , Vagus Nerve Stimulation , Adolescent , Adult , Aged , Female , Heart Rate , Humans , Male , Middle Aged , Retrospective Studies , Tinnitus/etiology , Tinnitus/physiopathology , Young AdultABSTRACT
Wound healing in the inner ear sensory epithelia is performed by the apical domains of supporting cells (SCs). Junctional F-actin belts of SCs are thin during development but become exceptionally thick during maturation. The functional significance of the thick belts is not fully understood. We have studied the role of F-actin belts during wound healing in the developing and adult cochlea of mice in vivo. We show that the thick belts serve as intracellular scaffolds that preserve the positions of surviving cells in the cochlear sensory epithelium. Junctions associated with the thick F-actin belts did not readily disassemble during wound healing. To compensate for this, basolateral membranes of SCs participated in the closure of surface breach. Because not only neighboring but also distant SCs contributed to wound healing by basolateral protrusions, this event appears to be triggered by contact-independent diffusible signals. In the search for regulators of wound healing, we inactivated RhoA in SCs, which, however, did not limit wound healing. RhoA inactivation in developing outer hair cells (OHCs) caused myosin II delocalization from the perijunctional domain and apical cell-surface enlargement. These abnormalities led to the extrusion of OHCs from the epithelium. These results demonstrate the importance of stability of the apical domain, both in wound repair by SCs and in development of OHCs, and that only this latter function is regulated by RhoA. Because the correct cytoarchitecture of the cochlear sensory epithelium is required for normal hearing, the stability of cell apices should be maintained in regenerative and protective interventions.
Subject(s)
Cochlea , Cytoskeleton/metabolism , Gene Expression Regulation, Developmental/genetics , Hair Cells, Vestibular/metabolism , Wound Healing/genetics , rhoA GTP-Binding Protein/metabolism , Actins/metabolism , Age Factors , Animals , Animals, Newborn , Cochlea/cytology , Cochlea/embryology , Cochlea/growth & development , Embryo, Mammalian , Epithelium/embryology , Epithelium/growth & development , Epithelium/metabolism , Epithelium/ultrastructure , Female , Hair Cells, Vestibular/ultrastructure , Mice , Mice, Inbred C57BL , Mice, Transgenic , Microscopy, Electron, Scanning , Myosin Type II/metabolism , Pregnancy , Receptor, Fibroblast Growth Factor, Type 3/genetics , Receptor, Fibroblast Growth Factor, Type 3/metabolism , rhoA GTP-Binding Protein/geneticsABSTRACT
Prevention of auditory hair cell death offers therapeutic potential to rescue hearing. Pharmacological blockade of JNK/c-Jun signaling attenuates injury-induced hair cell loss, but with unsolved mechanisms. We have characterized the c-Jun stress response in the mouse cochlea challenged with acoustic overstimulation and ototoxins, by studying the dynamics of c-Jun N-terminal phosphorylation. It occurred acutely in glial-like supporting cells, inner hair cells, and the cells of the cochlear ion trafficking route, and was rapidly downregulated after exposures. Notably, death-prone outer hair cells lacked c-Jun phosphorylation. As phosphorylation was triggered also by nontraumatic noise levels and none of the cells showing this activation were lost, c-Jun phosphorylation is a biomarker for cochlear stress rather than an indicator of a death-prone fate of hair cells. Preconditioning with a mild noise exposure before a stronger traumatizing noise exposure attenuated the cochlear c-Jun stress response, suggesting that the known protective effect of sound preconditioning on hearing is linked to suppression of c-Jun activation. Finally, mice with mutations in the c-Jun N-terminal phosphoacceptor sites showed partial, but significant, hair cell protection. These data identify the c-Jun stress response as a paracrine mechanism that mediates outer hair cell death.
Subject(s)
Biomarkers/metabolism , Hair Cells, Vestibular/metabolism , Hearing Loss, Noise-Induced/pathology , JNK Mitogen-Activated Protein Kinases/metabolism , Vestibulocochlear Nerve Injuries/pathology , Animals , Animals, Newborn , Apoptosis , Cell Death/drug effects , Cell Death/physiology , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Disease Models, Animal , Female , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Kanamycin/toxicity , Male , Mice , Mice, Inbred CBA , Mice, Transgenic , Noise/adverse effects , Protein Synthesis Inhibitors/toxicity , Time Factors , Transcription Factors/genetics , Transcription Factors/metabolism , Vestibulocochlear Nerve Injuries/chemically inducedABSTRACT
Hair cells of the organ of Corti (OC) of the cochlea exhibit distinct planar polarity, both at the tissue and cellular level. Planar polarity at tissue level is manifested as uniform orientation of the hair cell stereociliary bundles. Hair cell intrinsic polarity is defined as structural hair bundle asymmetry; positioning of the kinocilium/basal body complex at the vertex of the V-shaped bundle. Consistent with strong apical polarity, the hair cell apex displays prominent actin and microtubule cytoskeletons. The Rho GTPase Cdc42 regulates cytoskeletal dynamics and polarization of various cell types, and, thus, serves as a candidate regulator of hair cell polarity. We have here induced Cdc42 inactivation in the late-embryonic OC. We show the role of Cdc42 in the establishment of planar polarity of hair cells and in cellular patterning. Abnormal planar polarity was displayed as disturbances in hair bundle orientation and morphology and in kinocilium/basal body positioning. These defects were accompanied by a disorganized cell-surface microtubule network. Atypical protein kinase C (aPKC), a putative Cdc42 effector, colocalized with Cdc42 at the hair cell apex, and aPKC expression was altered upon Cdc42 depletion. Our data suggest that Cdc42 together with aPKC is part of the machinery establishing hair cell planar polarity and that Cdc42 acts on polarity through the cell-surface microtubule network. The data also suggest that defects in apical polarization are influenced by disturbed cellular patterning in the OC. In addition, our data demonstrates that Cdc42 is required for stereociliogenesis in the immature cochlea.
ABSTRACT
BACKGROUND: Ears of Brn3c null mutants develop immature hair cells, identifiable only by certain molecular markers, and undergo apoptosis in neonates. This partial development of hair cells could lead to enough neurotrophin expression to sustain sensory neurons through embryonic development. We have therefore investigated in these mutants the patterns of innervation and of expression of known neurotrophins. RESULTS: At birth there is a limited expression of BDNF and NT-3 in the mutant sensory epithelia and DiI tracing shows no specific reduction of afferents or efferents that resembles neurotrophin null mutations. At postnatal day 7/8 (P7/8), innervation is severely reduced both qualitatively and quantitatively. 1% of myosin VIIa-positive immature hair cells are present in the mutant cochlea, concentrated in the base. Around 20% of immature hair cells exist in the mutant vestibular sensory epithelia. Despite more severe loss of hair cells (1% compared to 20%), the cochlea retains many more sensory neurons (46% compared to 15%) than vestibular epithelia. Even 6 months old mutant mice have some fibers to all vestibular sensory epithelia and many more to the cochlear apex which lacks MyoVIIa positive hair cells. Topologically organized central cochlea projections exist at least until P8, suggesting that functional hair cells are not required to establish such projections. CONCLUSION: The limited expression of neurotrophins in the cochlea of Brn3c null mice suffices to support many sensory neurons, particularly in the cochlea, until birth. The molecular nature of the long term survival of apical spiral neurons remains unclear.
Subject(s)
Ear, Inner/innervation , Ear, Inner/pathology , Gene Expression Regulation, Developmental/genetics , Hair Cells, Auditory, Inner/metabolism , Transcription Factor Brn-3C/deficiency , Age Factors , Animals , Animals, Newborn , Brain-Derived Neurotrophic Factor/metabolism , Carbocyanines , Cell Count/methods , Dyneins/metabolism , Ear, Inner/growth & development , Embryo, Mammalian , Homeodomain Proteins , Immunohistochemistry/methods , In Situ Hybridization/methods , Mice , Mice, Knockout , Microscopy, Confocal/methods , Myosin VIIa , Myosins/metabolism , Neurons, Afferent/metabolism , Neurons, Afferent/physiology , Neurotrophin 3/metabolism , RNA, Messenger/metabolism , Spiral Ganglion/cytologyABSTRACT
The ototoxic action of aminoglycoside antibiotics leading to the loss of hair cells of the inner ear is well documented. However, the molecular mechanisms are poorly defined. We have previously shown that in neomycin-exposed organotypic cultures of the cochlea, the c-Jun N-terminal kinase (JNK) pathway--associated with stress, injury and apoptosis--is activated in hair cells and leads to their death. We have also shown that hair cell death can be attenuated by CEP-1347, an inhibitor of JNK signalling [Pirvola et al., J. Neurosci. 20 (2000) 43-50]. In the present study, we demonstrate that gentamicin-induced ototoxicity leads to JNK activation and apoptosis in the inner ear hair cells in vivo. We also show that systemic administration of CEP-1347 attenuates gentamicin-induced decrease of auditory sensitivity and cochlear hair cell damage. In addition, CEP-1347 treatment reduces the extent of hair cell loss in the ampullary cristae after gentamicin intoxication. Particularly, the inner hair cells of the cochlea and type I hair cells of the vestibular organs are protected. We have previously shown that also acoustic overstimulation leads to apoptosis of cochlear hair cells and that CEP-1347 can attenuate noise-induced sensory cell loss. These results suggest that activation of the JNK cascade may be a common molecular outcome of cellular stress in the inner ear sensory epithelia, and that attenuation of the lesion can be provided by inhibiting JNK activation.