Thymoquinone ameliorates age-related hearing loss in C57BL/6J mice by modulating Sirt1 activity and Bak1 expression.

Age-related hearing loss (AHL) is the most common sensory disorder of aged population. Currently, one of the most important sources of experimental medicine for AHL is medicinal plants. This study performed the first investigation of the effect of thymoquinone (TQ), a potent antioxidant, on AHL. Here, we used inbred C57BL/6J mice (B6 mice) as a successful experimental model of the early onset of AHL. The behavioral assessment of hearing revealed that the injection of a high dose of TQ (40 mg/kg; TQ40) significantly improved the auditory sensitivity of B6 mice at all tested frequencies (8, 16 and 22 kHz). Histological sections of cochlea from B6 mice injected with a low dose (20 mg/kg; TQ20) and high dose showed relatively less degenerative signs in the modiolus, hair cells and spiral ligaments, the main constituents of the cochlea. In addition, TQ40 completely restored the normal pattern of hair cells in B6 mice, as shown in scanning electron micrographs. Our data indicated that TQ20 and TQ40 reduced levels of Bak1-mediated apoptosis in the cochlea of B6 mice. Interestingly, the level of Sirt1, a positive regulator of autophagy, was significantly increased in B6 mice administered TQ40. In conclusion, TQ relieves the symptoms of AHL by downregulating Bak1 and activating Sirt1 in the cochlea of B6 mice.

Expression of Na/K-ATPase subunits in the human cochlea: a confocal and super-resolution microscopy study with special reference to auditory nerve excitation and cochlear implantation.

Background: For the first time the expression of the ion transport protein sodium/potassium-ATPase and its isoforms was analyzed in the human cochlea using light- and confocal microscopy as well as super-resolution structured illumination microscopy. It may increase our understanding of its role in the propagation and processing of action potentials in the human auditory nerve and how electric nerve responses are elicited from auditory prostheses. Material and methods: Archival human cochlear sections were obtained from trans-cochlear surgeries. Antibodies against the Na/K-ATPase ß1 isoform together with α1 and α3 were used for immunohistochemistry. An algorithm was applied to assess the expression in various domains. Results: Na/K ATPase ß1 subunit was expressed, mostly combined with the α1 isoform. Neurons expressed the ß1 subunit combined with α3, while satellite glial cells expressed the α1 isoform without recognized association with ß1. Types I and II spiral ganglion neurons and efferent fibers expressed the Na/K-ATPase α3 subunit. Inner hair cells, nerve fibers underneath, and efferent and afferent fibers in the organ of Corti also expressed α1. The highest activity of Na/K-ATPase ß1 was at the inner hair cell/nerve junction and spiral prominence. Conclusion: The human auditory nerve displays distinct morphologic features represented in its molecular expression. It was found that electric signals generated via hair cells may not go uninterrupted across the spiral ganglion, but are locally processed. This may be related to particular filtering properties in the human acoustic pathway.

Spontaneous otoacoustic emissions in teiid lizards.

SOAE from the last major lizard family not yet systematically investigated, the teiids, were collected from the genera Callopistes, Tupinambis and Cnemidophorus. Although their papillae show characteristics of the family Teiidae, the papillae differ both in their size and in the arrangement of uni- and bi-directional hair-cell areas. Among these three genera, Callopistes showed few (2 or 3) SOAE peaks, whereas the other two genera showed more (up to 6 per ear). In the absence of knowledge of the tonotopic maps, however, it was not possible to clearly relate the spectral patterns to the differences in papillar anatomy, suggesting that the determinants of these patterns may be more subtle than anticipated.

Ultrastructural Identification and Colocalization of Interacting Proteins in the Murine Cochlea by Post-Embedding Immunogold Transmission Electron Microscopy.

Verification of the presence and location of a protein within tissue can be accomplished by western blotting and immunohistochemistry, using either paraffin or frozen sections. Affinity purification by reciprocal coimmunoprecipitations using the tissue of interest can demonstrate the existence of an interacting pair of proteins. Ultimately, the ability to visualize the interaction at the cellular level is desired. Precise location(s) of interacting proteins in situ can be accomplished by ultrastructural localization with high-quality primary antibodies and small-particle-size Au-conjugated secondary antibodies. Visualization can be obtained with a transmission electron microscope fitted with a high-resolution camera permitting magnifications that exceed 2 × 10(5), and, to date, resolution capability of 20+ Mpixels, thus enabling localization of the target protein to within nanometers of the actual location. Here, we report the method by which immunolocalization at the level of the electron microscope is accomplished using the post-embedding technique, i.e., performing antibody labeling of proteins on ultrathin sections of tissue embedded in acrylic resin.

Association of Caffeine and Hearing Recovery After Acoustic Overstimulation Events in a Guinea Pig Model.

IMPORTANCE: Noise-induced hearing loss is an increasingly worrisome problem. Although caffeine intake is common in people involved in noise-related environments, the effect of caffeine on the recovery of hearing after a temporary threshold shift requires further understanding. OBJECTIVES: To determine whether caffeine impairs hearing recovery in a guinea pig model exposed to acoustic overstimulation. DESIGN, SETTING, AND SUBJECTS: This experiment at the McGill University Auditory Sciences Laboratory used 24 female albino guinea pigs (age, 6 months; weight, 500-600 g) divided randomly into 3 groups of 8 animals each. Group 1 was exposed to caffeine; group 2, acoustic overstimulation events (AOSEs); and group 3, both. Data were collected from July 1, 2013, to March 30, 2014, and analyzed from April 1 to August 1, 2014. INTERVENTIONS: Daily caffeine dose for groups 1 and 3 consisted of 25 mg/kg administered intraperitoneally for 15 days. The AOSEs were administered on days 1 and 8 and consisted of 1 hour of 110-dB pure-tone sound. MAIN OUTCOMES AND MEASURES: Serial auditory brainstem response (ABR) tests to determine the audiological threshold shift and recovery were obtained at baseline and on days 1 (1 hour after the first AOSE), 4, 8 (before and 1 hour after the second AOSE), 11, and 15. Scanning electron and light microscopy of the cochleas were performed to determine morphologic changes. RESULTS: The day 1 post-AOSE measurement resulted in a similar threshold shift in all animals in groups 2 and 3 at all frequencies tested (8, 16, 20, and 25 kHz). The maximum threshold shift was at 16 kHz, with a mean of 66.12 dB. By day 8, the threshold shift in group 2 recovered completely at all frequencies except 20 kHz, where a mean threshold shift of 20.63 dB of sound pressure level (SPL) was present. Hearing impairment in group 3 persisted in 8-, 16-, and 25-kHz frequencies with thresholds of 21.88, 28.13, and 26.25 dB SPL, respectively (P = .001). After a second AOSE at day 8, similar threshold shift and outcome were recorded on day 15 compared with day 8, with a mean threshold shift at 20 kHz of 29.38 dB SPL in group 2 and mean threshold shifts at 8, 16, 20, and 25 kHz of 29.38, 35.63, 40.63, and 38.75 dB SPL, respectively, in group 3. The difference in ABR threshold recovery was in concordance with scanning electronic and light microscopy findings for each group. CONCLUSIONS AND RELEVANCE: A daily dose of caffeine was found to impair the recovery of hearing after an AOSE.

Thyroid hormone is required for pruning, functioning and long-term maintenance of afferent inner hair cell synapses.

Functional maturation of afferent synaptic connections to inner hair cells (IHCs) involves pruning of excess synapses formed during development, as well as the strengthening and survival of the retained synapses. These events take place during the thyroid hormone (TH)-critical period of cochlear development, which is in the perinatal period for mice and in the third trimester for humans. Here, we used the hypothyroid Snell dwarf mouse (Pit1(dw)) as a model to study the role of TH in afferent type I synaptic refinement and functional maturation. We observed defects in afferent synaptic pruning and delays in calcium channel clustering in the IHCs of Pit1(dw) mice. Nevertheless, calcium currents and capacitance reached near normal levels in Pit1(dw) IHCs by the age of onset of hearing, despite the excess number of retained synapses. We restored normal synaptic pruning in Pit1(dw) IHCs by supplementing with TH from postnatal day (P)3 to P8, establishing this window as being critical for TH action on this process. Afferent terminals of older Pit1(dw) IHCs showed evidence of excitotoxic damage accompanied by a concomitant reduction in the levels of the glial glutamate transporter, GLAST. Our results indicate that a lack of TH during a critical period of inner ear development causes defects in pruning and long-term homeostatic maintenance of afferent synapses.

Noise induced reversible changes of cochlear ribbon synapses contribute to temporary hearing loss in mice.

CONCLUSION: Noise exposure can cause a decline in cochlear ribbon synapses and result in consequent hearing loss. The reduction of synaptic puncta appears reversible and may contribute to hearing restoration in mice after noise exposure. OBJECTIVE: To detect whether noise induced reversible changes of cochlear ribbon synapses contribute to temporary hearing loss in C57BL/6J mice. METHODS: The mice were assigned randomly to five groups and exposed to white noise at 110 dB SPL for 2 h except the control group. ABR thresholds were acquired before noise exposure (control), immediately following exposure (Day 0), or on Days 4, 7, or 14 after noise exposure. Light microscopy, scanning emission microscopy, and whole mounts examination was utilized to study whether there is morphology change of outer hair cells (OHC), inner hair cells (IHC), or spiral ganglion cells (SGN) due to the 110 dB white noise. Moreover, experimental approaches, including immunostaining and confocal microcopy, were used to detect whether ribbon synapses were the primary targets of noise-induced temporary hearing loss. RESULT: Exposure to 110 dB white noise for 2 h induced TTS in mice, with the maximal ABR threshold elevations seen on the 4(th) day after noise exposure. There were no significant morphological changes in the cochlea. Paralleled changes of pre-synaptic ribbons in both the number and post-synaptic density (PSDs) during this noise exposure were detected. The number of pre-synaptic ribbon, post-synaptic density (PSDs), and co-localized puncta correlated with the shifts of ABR thresholds. Moreover, a complete recovery of ABR thresholds and synaptic puncta was seen on the 14(th) day after the noise stimulations.

Auto-inflammatory challenge of the endolymphatic sac--Cochlear damage measured by distortion product oto-acoustic emissions.

CONCLUSION: Twenty-five rats were challenged by an immunologic attack of the endolymphatic sac. After 6 months, distortion product oto-acoustic emissions (DPOAE) revealed a dysfunction of the outer hair cells and immunological active cells were observed in the endolymphatic sac. This information could contribute to the understanding of Ménière's disease. OBJECTIVES: This study investigated if an autoimmune challenge of the endolymphatic sac could affect DPOAE output measurements in rats. Also, a potential autoimmune cell infiltration of the endolymphatic sac was investigated. METHODS: Eighteen Lewis rats were immunized with a crude endolymphatic sac extract in complete Freund's adjuvant. Seven control animals were injected with Freund's adjuvant in saline. Cochlear damage was estimated by DPOAE dynamics 3 weeks and 6 months after the immunization. Infiltrative cells in the endolymphatic sac were investigated with transmission electron microscopy. RESULTS: The hearing assessment 6 months after immunization revealed a reduction of the DPOAE, on the full range of frequencies (2-63 kHz) in an average of the mean, of 2 dB ± 1.1 in the immunized group compared to the controls (p < 0.05). The same test showed a 2.5 dB decrease from 2 to 5 kHz (p < 0.01). Immunological active cells were observed in the endolymphatic sac in most of the immunized rats.

Electron microscopic examination of effects of bogma raki and walnut on cochlea: an experimental study.

Illegal alcohol beverages known as bogma raki in our country are consumed widely in our region. The studies investigating the relationship between alcohol consumption and hearing ability report different results. In this study, we aimed to investigate the toxic effects of bogma raki that contains neurotoxic substances on cochlea by electron microscopy. To the best of our knowledge, this study is the first in the literature. A total of 48 Wistar male albino rats (aged 12-16 weeks and weighing 200-240 g) were used in the study. The rats were divided into 4 groups with 12 animals in each group. The groups include control, bogma raki, walnut, and walnut + bogma raki groups. Bogma raki (30% v/v, 9.2 ml kg(-1) day(-1)) is added to drinking water of rats in bogma raki group (n = 12) for 4 weeks. Walnut group rats (n = 12) are fed with standard rat food and walnut without limitation (10 g kg(-1) day(-1)). Bogma raki + walnut group rats (n = 12) are fed with standard rat food and walnut and bogma raki is added to drinking water. The cochleas were dissected and removed en bloc and examined by electron microscopy. Perineuronal oedema around neurons of spiral ganglion and hairy cells of organ of Corti were present in the bogma raki group, walnut group and bogma raki + walnut group under electron microscopic examination. Comparing these three groups, there were no differences in the ultrastructural pathological changes. In the ultrastructural examination of the myelinated axons forming cochlear nerve, no ultrastructural pathology was detected in all the groups.

[Lack of protection against gentamicin ototoxicity by auditory conditioning with noise]. / Falta de proteção contra a ototoxicidade da gentamicina pelo condicionamentoauditivo com ruído.

INTRODUCTION: Auditory conditioning consists of the pre-exposure to low levels of a potential harmful agent to protect against a subsequent harmful exposure. OBJECTIVE: To confirm if conditioning with an agent different from that used to cause the trauma can also be effective. METHODS: This was an experimental study with 17 guinea pigs, divided into three groups: an ototoxic control group (Cont) that received intramuscular administration of gentamicin 160 mg/kg/day for ten consecutive days, but no sound exposure; a sound control group (Sound) that was exposed to 85 dB broadband noise centered at 4 kHz, 30 min each day for ten consecutive days, but received no ototoxic medications; and an experimental group (Expt) that received sound exposure identical to the Sound group and after each noise presentation, received gentamicin similarly to Cont group. The animals were evaluated by distortion product otoacoustic emissions (DPOAEs), brainstem auditory evoked potentials (BAEPs), and scanning electron microscopy. RESULTS: The animals that were conditioned with noise did not show any protective effect compared with the ones that received only the ototoxic gentamicin administration. This lack of protection was observed functionally and morphologically. CONCLUSION: Conditioning with 85 dB broadband noises, 30 min a day for ten consecutive days does not protect against an ototoxic gentamicin administration of 160 mg/kg/day for ten consecutive days in the guinea pig.

Lack of protection against gentamicin ototoxicity by auditory conditioning with noise / Falta de proteção contra a ototoxicidade da gentamicina pelo condicionamento auditivo com ruído

INTRODUCTION: Auditory conditioning consists of the pre-exposure to low levels of a potential harmful agent to protect against a subsequent harmful presentation. OBJECTIVE: To confirm if conditioning with an agent different from the used to cause the trauma can also be effective. METHOD: Experimental study with 17 guinea pigs divided as follows: group Som: exposed to 85 dB broadband noise centered at 4 kHz, 30 minutes a day for 10 consecutive days; group Cont: intramuscular administration of gentamicin 160 mg/kg a day for 10 consecutive days; group Expt: conditioned with noise similarly to group Som and, after each noise presentation, received gentamicin similarly to group Cont. The animals were evaluated by distortion product otoacoustic emissions (DPOAEs), brainstem auditory evoked potentials (BAEPs) and scanning electron microscopy. RESULTS: The animals that were conditioned with noise did not show any protective effect compared to the ones that received only the ototoxic gentamicin administration. This lack of protection was observed functionally and morphologically. CONCLUSION: Conditioning with 85 dB broadband noise, 30 min a day for 10 consecutive days does not protect against an ototoxic gentamicin administration of 160 mg/kg a day for 10 consecutive days in the guinea pig. .

INTRODUÇÃO: O condicionamento auditivo consiste da pré-exposição de um agente lesivo em baixos níveis para proteger contra uma posterior apresentação lesiva. OBJETIVO: Confirmar se o condicionamento com um agente diferente do utilizado para causar o trauma pode ser efetivo. MÉTODO: Estudo experimental com 17 cobaias albinas divididas como a seguir- grupo Som: exposto a um ruído branco de 85 dB centrado em 4 kHz, 30 minutos por dia, por 10 dias consecutivos; grupo Cont: administração intramuscular de gentamicina 160 mg/kg por dia, por 10 dias consecutivos; grupo Expt: condicionado com ruído como o grupo Som. Após cada exposição ao ruído, recebeu gentamicina similarmente ao grupo Cont. Os animais foram avaliados por emissões otoacústicas produto de distorção (EOAPDs), potencial evocado auditivo de tronco encefálico (PEATE) e microscopia eletrônica de varredura (MEV). RESULTADOS: Os animais que foram condicionados com ruído não mostraram qualquer efeito protetor quando comparados com os que receberam apenas a gentamicina em doses ototóxicas. Esta ausência de proteção foi observada tanto funcionalmente quanto morfologicamente. CONCLUSÃO: Os autores concluíram que o condicionamento com ruído branco a 85 dB por 30 minutos, por dia por 10 dias consecutivos, não protege contra uma administração de gentamicina 160 mg/kg/dia, por 10 dias consecutivos. .

Attenuation of noise-induced hearing loss using methylene blue.

The overproduction of reactive oxygen species (ROS) and reactive nitrogen species (RNS) has been known to contribute to the pathogenesis of noise-induced hearing loss. In this study, we discovered that in BALB/c mice pretreatment with methylene blue (MB) for 4 consecutive days significantly protected against cochlear injury by intense broad-band noise for 3 h. It decreased both compound threshold shift and permanent threshold shift and, further, reduced outer hair cell death in the cochlea. MB also reduced ROS and RNS formation after noise exposure. Furthermore, it protected against rotenone- and antimycin A-induced cell death and also reversed ATP generation in the in vitro UB-OC1 cell system. Likewise, MB effectively attenuated the noise-induced impairment of complex IV activity in the cochlea. In addition, it increased the neurotrophin-3 (NT-3) level, which could affect the synaptic connections between hair cells and spiral ganglion neurons in the noise-exposed cochlea, and also promoted the conservation of both efferent and afferent nerve terminals on the outer and inner hair cells. These findings suggest that the amelioration of impaired mitochondrial electron transport and the potentiation of NT-3 expression by treatment with MB have a significant therapeutic value in preventing ROS-mediated sensorineural hearing loss.

Tonotopic morphometry of the lamina reticularis of the guinea pig cochlea with associated microstructures and related mechanical implications.

Morphometry of the lamina reticularis of the guinea pig cochlea was performed using scanning electron microscopy. Seventy-four geometrical parameters of the lamina reticularis, the bundles of stereocilia, and individual stereocilia, in all rows of hair cells and within the individual hair cells, were measured at ten equally spaced locations along the longitudinal direction of the cochlea. Variations of the parameters versus the longitudinal coordinate were statistically analyzed and fitted with polynomials (constant, linear, or quadratic). Our data show that a unique set of geometrical parameters of inner and outer hair cells is typical for every frequency-dependent position at the lamina reticularis. Morphology of the outer hair cell structures varies more than respective parameters of the inner hair cells. Mechanical modeling using the obtained geometrical parameters provides a novel glance at the mechanical characteristics with respect to the cochlear tonotopy.

Otoprotection in guinea pigs exposed to pesticides and ginkgo biloba.

UNLABELLED: Pesticides are widely used in agriculture, despite the risk of hearing loss related to the exposure to their chemical components. This study looks into protective drugs to counteract the ototoxicity of pesticides. OBJECTIVE: This study aims to analyze the effect ginkgo biloba extract may have in protecting against possible cochlear damage caused by organophosphate pesticides (methamidophos). Anatomic changes are assessed through surface and electron microscopy. MATERIALS AND METHODS: This is a prospective experimental study. Twenty-one guinea pigs were given saline solution, pesticide, and ginkgo biloba alone or combined for seven consecutive days. Then their cochleas were removed and examined in a scanning electron microscope. RESULTS: Pesticide-exposed guinea pigs had morphological alterations in their cochleas and injuries in the three turns analyzed through electron microscopy. Injury intensity varied according to the dosages of the agents given to the test subjects. Guinea pigs treated with pesticide and ginkgo biloba maintained the architecture of their outer hair cells in all cochlear turns. CONCLUSION: The antioxidant properties found in the ginkgo biloba extract protected guinea pigs from pesticide ototoxicity.

Otoproteção em cobaias expostas a agrotóxico e ginkgo biloba / Otoprotection in guinea pigs exposed to pesticides and ginkgo biloba

Os agrotóxicos são amplamente utilizados na agricultura e, atualmente, fazem parte do grupo de agentes químicos que podem levar à perda auditiva. A identificação de drogas que, associadas aos ototóxicos, possam atuar como otoprotetores é objeto de estudo. OBJETIVO: Analisar a existência de efeito otoprotetor do extrato de Ginkgo biloba aos possíveis danos cocleares causados pelo agrotóxico do grupo dos organofosforados - metamidofós, avaliando-se as alterações anatômicas por meio da microscopia eletrônica de superfície. MATERIAL E MÉTODO: Estudo experimental prospectivo utilizando 21 cobaias, que sofreram ação da administração de soro fisiológico, agrotóxico e ginkgo biloba isoladamente e associadas, durante sete dias consecutivos. Após, as cócleas foram removidas e avaliadas anatomicamente pela microscopia eletrônica de varredura. RESULTADOS: As cobaias submetidas ao agrotóxico apresentaram alterações morfológicas cocleares, com lesões nas três espiras analisadas na microscopia eletrônica, intensificadas de acordo com a dosagem recebida do agente. As cobaias tratadas com agrotóxico e Ginkgo biloba apresentaram uma manutenção da arquitetura ciliar nas células ciliadas externas em todas as espiras da cóclea. CONCLUSÃO: O extrato de Ginkgo biloba, por sua ação antioxidante, atuou como fator otoprotetor à ototoxicidade pelo agrotóxico em cobaias.

Pesticides are widely used in agriculture, despite the risk of hearing loss related to the exposure to their chemical components. This study looks into protective drugs to counteract the ototoxicity of pesticides. OBJECTIVE: This study aims to analyze the effect ginkgo biloba extract may have in protecting against possible cochlear damage caused by organophosphate pesticides (methamidophos). Anatomic changes are assessed through surface and electron microscopy. MATERIALS AND METHODS: This is a prospective experimental study. Twenty-one guinea pigs were given saline solution, pesticide, and ginkgo biloba alone or combined for seven consecutive days. Then their cochleas were removed and examined in a scanning electron microscope. RESULTS: Pesticide-exposed guinea pigs had morphological alterations in their cochleas and injuries in the three turns analyzed through electron microscopy. Injury intensity varied according to the dosages of the agents given to the test subjects. Guinea pigs treated with pesticide and ginkgo biloba maintained the architecture of their outer hair cells in all cochlear turns. CONCLUSION: The antioxidant properties found in the ginkgo biloba extract protected guinea pigs from pesticide ototoxicity.

Temporal aspects of suppression in distortion-product otoacoustic emissions.

This study examined the time course of cochlear suppression using a tone-burst suppressor to measure decrement of distortion-product otoacoustic emissions (DPOAEs). Seven normal-hearing subjects with ages ranging from 19 to 28 yr participated in the study. Each subject had audiometric thresholds ≤ 15 dB HL [re ANSI (2004) Specifications for Audiometers] for standard octave and inter-octave frequencies from 0.25 to 8 kHz. DPOAEs were elicited by primary tones with f(2) = 4.0 kHz and f(1) = 3.333 kHz (f(2)/f(1) = 1.2). For the f(2), L(2) combination, suppression was measured for three suppressor frequencies: One suppressor below f(2) (3.834 kHz) and two above f(2) (4.166 and 4.282 kHz) at three levels (55, 60, and 65 dB SPL). DPOAE decrement as a function of L(3) for the tone-burst suppressor was similar to decrements obtained with longer duration suppressors. Onset- and setoff- latencies were ≤ 4 ms, in agreement with previous physiological findings in auditory-nerve fiber studies that suggest suppression results from a nearly instantaneous compression of the waveform. Persistence of suppression was absent for the below-frequency suppressor (f(3) = 3.834 kHz) and was ≤ 3 ms for the two above-frequency suppressors (f(3) = 4.166 and 4.282 kHz).

Distribution of standing-wave errors in real-ear sound-level measurements.

Standing waves can cause measurement errors when sound-pressure level (SPL) measurements are performed in a closed ear canal, e.g., during probe-microphone system calibration for distortion-product otoacoustic emission (DPOAE) testing. Alternative calibration methods, such as forward-pressure level (FPL), minimize the influence of standing waves by calculating the forward-going sound waves separate from the reflections that cause errors. Previous research compared test performance (Burke et al., 2010) and threshold prediction (Rogers et al., 2010) using SPL and multiple FPL calibration conditions, and surprisingly found no significant improvements when using FPL relative to SPL, except at 8 kHz. The present study examined the calibration data collected by Burke et al. and Rogers et al. from 155 human subjects in order to describe the frequency location and magnitude of standing-wave pressure minima to see if these errors might explain trends in test performance. Results indicate that while individual results varied widely, pressure variability was larger around 4 kHz and smaller at 8 kHz, consistent with the dimensions of the adult ear canal. The present data suggest that standing-wave errors are not responsible for the historically poor (8 kHz) or good (4 kHz) performance of DPOAE measures at specific test frequencies.

Effects of hyperbaric oxygen treatment on auditory hair cells after acute noise damage.

Acute acoustic trauma (AAT) is a sudden sensorineural hearing loss caused by exposure of the hearing organ to acoustic overstimulation, typically an intense sound impulse, hyperbaric oxygen therapy (HOT), which favors repair of the microcirculation, can be potentially used to treat it. Hence, this study aimed to assess the effects of HOT on guinea pigs exposed to acoustic trauma. Fifteen guinea pigs were exposed to noise in the 4-kHz range with intensity of 110 dB sound level pressure for 72 h. They were assessed by brainstem auditory evoked potential (BAEP) and by distortion product otoacoustic emission (DPOAE) before and after exposure and after HOT at 2.0 absolute atmospheres for 1 h. The cochleae were then analyzed using scanning electron microscopy (SEM). There was a statistically significant difference in the signal-to-noise ratio of the DPOAE amplitudes for the 1- to 4-kHz frequencies and the SEM findings revealed damaged outer hair cells (OHC) after exposure to noise, with recovery after HOT (p = 0.0159), which did not occur on thresholds and amplitudes to BAEP (p = 0.1593). The electrophysiological BAEP data did not demonstrate effectiveness of HOT against AAT damage. However, there was improvement of the anatomical pattern of damage detected by SEM, with a significant reduction of the number of injured cochlear OHC and their functionality detected by DPOAE.

Expression and localization of 11beta-hydroxysteroid dehydrogenase (11betaHSD) in the rat endolymphatic sac.

CONCLUSIONS: 11beta-Hydroxysteroid dehydrogenase type 2 (11bHSD-2) enables aldosterone to bind to mineralocorticoid receptors (MRs) selectively by converting cortisol (corticosterone) into inactive metabolites. Its expression in the endolymphatic sac (ES) suggests that aldosterone may selectively act on the ES through its binding to MRs by the action of 11betaHSD-2, and supports the notion that ES is an aldosterone target organ. We propose that 11betaHSD-2 is a dominant isoform of 11betaHSDs in the ES, and the ES (especially the intermediate portion of the ES) may be the main aldosterone target in the inner ear. OBJECTIVE: The purpose of this study was to examine 11bHSD isoform expression in the rat inner ear, mainly 11betaHSD-2 in the ES. MATERIALS AND METHODS: In the ES and whole cochlea, 11betaHSDs were examined by RT-PCR using highly specific ES RNA by laser capture microdissection. In addition, 11betaHSD-2 localization in the rat ES was determined by immunohistochemistry. RESULTS: RT-PCR demonstrated 11betaHSD-2 expressed in the rat ES. In addition, its localization was observed mainly in the intermediate portion and a faint immune positive signal was observed in other parts of the ES. In contrast, 11bHSD-1 was undetectable in the ES by RT-PCR. Both types of 11betaHSDs were expressed in rat cochlea.

Smad5 haploinsufficiency leads to hair cell and hearing loss.

The Smads are a group of related intracellular proteins critical for transmitting the signals to the nucleus from the transforming growth factor-beta superfamily at the cell surface. Knockout of the Smad5 is embryonic lethal. However, the Smad5 knockout of single allele (+/-) could survive. We used Smad5 heterozygous knockout (+/-) to determine the role of Smad5 in the development of inner ear morphology and function. In situ hybridization showed that Smad5 was expressed predominantly in hair cells, spiral ganglion, and supporting cells. Measurements of hearing thresholds using auditory brainstem response showed that Smad5 defect resulted in progressive hearing loss between 4 and 24 weeks after birth. Morphological examination revealed apoptosis in the inner ear, with significant loss of outer hair cells in adult Smad5 mutant mice. Our results indicated that deficiency in the Smad5-mediated signaling resulted in apoptosis of hair cells, suggesting Smad5 is a gene that may be related with presbycusis.