Anion Transport Proteins ; physiology ; Hair Cells, Auditory, Outer ; physiology ; Humans

BACKGROUND AND OBJECTIVES: This study aims to verify that one of the causes of tinnitus is the malfunction of outer hair cells and, on the basis of this, to investigate the usefulness of otoacoustic emissions by performing transient evoked otoacoustic emissions (TEOAE) and distor-tion product otoacoustic emissions (DPOAE). SUBJECTS AND METHOD: Included in the study were forty-one patients who had normal hearing in the range from 0.5 to 8 kHz, and complained of unilateral tinnitus. In these patients, hearing in bilateral ears, TEOAE, DPOAE, as well as the frequency & amplitude of their tinnitus were measured. RESULTS: No statistically significant difference was found in bilateral hearing in patients who complained of unilateral tinnitus. However, TEOAE and DPOAE showed a statistically significant difference with their p-values at 0.04 and 0.004, respectively. CONCLUSION: The results of this study suggested that TEOAE testing and DPOAE testing provide an important clue for verifying that the loss of outer hair cells contributed to the development of symptoms suffered by tinnitus patients with normal hearing.
Ear ; Hair Cells, Auditory, Outer ; Hearing* ; Humans ; Methods ; Tinnitus*

Inhibition of K⁺ outward currents by linopirdine in the outer hair cells (OHCs) of circling mice (homozygous (cir/cir) mice), an animal model for human deafness (DFNB6 type), was investigated using a whole cell patch clamp technique. Littermate heterozygous (+/cir) and ICR mice of the same age (postnatal day (P) 0 –P6) were used as controls. Voltage steps from –100 mV to 40 mV elicited small inward currents (–100 mV~–70 mV) and slow rising K⁺ outward currents (–60 mV ~40 mV) which activated near –50 mV in all OHCs tested. Linopirdine, a known blocker of K⁺ currents activated at negative potentials (I(K,n)), did cause inhibition at varying degree (severe, moderate, mild) in K⁺ outward currents of heterozygous (+/cir) or homozygous (cir/cir) mice OHCs in the concentration range between 1 and 100 µM, while it was apparent only in one ICR mice OHC out of nine OHCs at 100 µM. Although the half inhibition concentrations in heterozygous (+/cir) or homozygous (cir/cir) mice OHCs were close to those reported in I(K,n), biophysical and pharmacological properties of K⁺ outward currents, such as the activation close to –50 mV, small inward currents evoked by hyperpolarizing steps and TEA sensitivity, were not in line with I(K,n) reported in other tissues. Our results show that the delayed rectifier type K⁺ outward currents, which are not similar to I(K,n) with respect to biophysical and pharmacological properties, are inhibited by linopirdine in the developing (P0~P6) homozygous (cir/cir) or heterozygous (+/cir) mice OHCs.
Animals ; Deafness ; Hair Cells, Auditory, Outer* ; Humans ; Mice* ; Mice, Inbred ICR ; Models, Animal ; Tea

The cochlear auditory epithelium contains two types of sound receptors, inner hair cells (IHCs) and outer hair cells (OHCs). Mouse models for labelling juvenile and adult IHCs or OHCs exist; however, labelling for embryonic and perinatal IHCs or OHCs are lacking. Here, we generated a new knock-in Fgf8^P2A-3×GFP/+ (Fgf8^GFP/+) strain, in which the expression of a series of three GFP fragments is controlled by endogenous Fgf8 cis-regulatory elements. After confirming that GFP expression accurately reflects the expression of Fgf8, we successfully obtained both embryonic and neonatal IHCs with high purity, highlighting the power of Fgf8^GFP/+. Furthermore, our fate-mapping analysis revealed, unexpectedly, that IHCs are also derived from inner ear progenitors expressing Insm1, which is currently regarded as an OHC marker. Thus, besides serving as a highly favorable tool for sorting early IHCs, Fgf8^GFP/+ will facilitate the isolation of pure early OHCs by excluding IHCs from the entire hair cell pool.
Animals ; Mice ; Hair Cells, Auditory, Inner ; Cochlea/metabolism* ; Hair Cells, Auditory, Outer/metabolism* ; Disease Models, Animal ; Fibroblast Growth Factor 8/metabolism*

In order to elucidate the mechanism underlying the attenuation of streptomycin ototoxicity by tetramethylpyrazine (TMP), the present study investigated the effect of TMP on the outward K(+) current in the outer hair cells of guinea pig cochlea. Sixty guinea pigs were divided into 6 groups randomly. Auditory brainstem response (ABR) was used to observe the change in thresholds and to evaluate ototoxicity induced by streptomycin. Whole-cell patch-clamp technique was used to observe the effect of TMP on outward K(+) current in isolated outer hair cells. The results showed that TMP attenuated the threshold shift caused by streptomycin and increased the amplitudes of Ca(2+)-sensitive K(+) current [I(K(Ca))] in the outer hair cells. The present data suggest that TMP displays anti-ototoxicity induced by streptomycin. The augmented amplitudes of I(K(Ca)) of the outer hair cells induced by TMP may be one of the mechanisms underlying its ototoxicity-attenuating effect.
Animals ; Auditory Threshold ; Cochlea ; cytology ; Evoked Potentials, Auditory, Brain Stem ; Guinea Pigs ; Hair Cells, Auditory, Outer ; drug effects ; Patch-Clamp Techniques ; Potassium Channels ; metabolism ; Pyrazines ; Streptomycin ; toxicity

AIMThe comparison of several methods which were used to isolate cochlear outer hair cells and the observations of morphology were researched.

METHODSThree different separating methods of outer hair cells in cochlea were adopted; the morphology of outer hair cells in cochlea and the morphology of cochlear stretched preparation in Silver Nitrate staining were also investigated.

RESULTSSingle alive OHC in cochlea was disassociated by all methods, with microscope and cochlear stretched preparation's staining. We could also observe appearances and distributions of OHC in cochlea.

CONCLUSIONIt is successful to isolate single alive cochlear OHC, and it will be very important to investigate deeply normal physiological functions and changes of functions and morphology in some pathologic status in cochlear OHCs.

Animals ; Cell Separation ; methods ; Cochlea ; cytology ; physiology ; Female ; Guinea Pigs ; Hair Cells, Auditory, Outer ; cytology ; physiology ; Male ; Silver Staining

OBJECTIVETo explore the quantitative relationship between the reduction of distortion product otoacoustic emission (DPOAE) and the percentage of outer hair cell loss.

METHODSCoadministration of cisplatin (0.2 mg/kg) and ethacrynic acid (40 mg/kg) were used to establish a cochlear lesion model in chinchillas. DPOAE was measured before and 1 week, 2 weeks, and 3 weeks later respectively after cisplatin and ethacrynic acid treatment. Animals were terminated 3 weeks after the treatment. Cochlear surface preparations were performed, and the cochlear hair cells were counted through entire length of the cochlea. The correlation between DPOAE reduction and outer hair cell missing was analyzed using Pearson correlation analysis.

RESULTSCisplatin and ethacrynic acid treatment induced cochlear hair cell lesion that the outer hair cell loss in the cochlea developed in a stereotypic pattern; damage began in the base of the cochlea and progressed towards the apex. Reduction of DPOAE was relatively consistent with outer hair cells loss. On the average, 1% outer hair cells loss may result in 0.24 dB reduction in DPOAE levels. Pearson analysis showed a positive correlation between the reduction in DPOAE and missing of outer hair cells (r = 0.796, P < 0.05).

CONCLUSIONSIt may be helpful to evaluate missing percentage of outer hair cells from reduction in DPOAE levels.

Animals ; Cell Count ; Chinchilla ; Cisplatin ; Disease Models, Animal ; Ethacrynic Acid ; Hair Cells, Auditory, Outer ; cytology ; pathology ; Otoacoustic Emissions, Spontaneous

OBJECTIVETo investigate the pathway and mechanism of noise exposure induced out hair cells (OHC) apoptosis.

METHODSThe cochleae of control and noise exposure group were dissected. The activity of caspase 3, an important mediator of apoptosis, in OHC, was examined with carboxyfluorescein-labeled fluoromethyl ketone (FMK)-peptide inhibitors. The apoptosis inducing factor (AIF) translocation from mitochondria in OHC were further examined by immunohistology method. The nuclei were labeled with PI and the mitochondrion was labeled with Mito-tracker. Whole mount organ of Corti was prepared. Morphological and fluorescent change was observed use confocal microscope.

RESULTSIn the normal OHC, AIF is distributed where the mitochondria were located and no activated caspase 3 was observed. After the animals exposed to broadband noise at 122 dB in 4 h/day for 2 days, both apoptosis and necrosis were appeared in OHC. AIF translocated from mitochondrion to nuclei in apoptotic and necrotic OHC following noise exposure. The noise exposure triggered activation of caspase 3 in apoptic hair cells. But no caspase 3 activation appeared in necrotic OHC.

CONCLUSIONSThese findings indicated that the caspase-dependent pathway is an important pathway in noise exposure induced apoptosis. And AIF also involves OHC death pathway following noise exposure.

Animals ; Apoptosis ; Apoptosis Inducing Factor ; metabolism ; Caspase 3 ; metabolism ; Female ; Guinea Pigs ; Hair Cells, Auditory, Outer ; metabolism ; pathology ; Male ; Noise

OBJECTIVE: We used electrophysiological methods to study that whether GABA could elicit OHCs outward currents provide evidence for exsitence of GABA-A receptor and investige the relationship between the effect of GABA and the development of OHCs. METHOD: We used whole-cell recording OHCs at current-clamp or voltage-clamp to verify the function of GABA receptor on OHCs. Then we counteds the responsive cells vs. total number cells, and according to results to study the relationships between the GABA receptor and development of OHCs. RESULT: OHC was elicited outward current or hyperpolarized by GABA and the responsive cells were decreased with development. CONCLUSION The result of GABA receptor decreasing with development suggested that the receptor may draw efferents to OHCs or facilitate the MOC-OHC synapse formation.
Animals ; Electrophysiological Phenomena ; Hair Cells, Auditory, Outer ; physiology ; Patch-Clamp Techniques ; Rats ; Receptors, GABA-A ; physiology ; gamma-Aminobutyric Acid ; physiology

In mammals, the piezoelectric protein, Prestin, endows the outer hair cells (OHCs) with electromotility (eM), which confers the capacity to change cellular length in response to alterations in membrane potential. Together with basilar membrane resonance and possible stereociliary motility, Prestin-based OHC eM lays the foundation for enhancing cochlear sensitivity and frequency selectivity. However, it remains debatable whether Prestin contributes to ultrahigh-frequency hearing due to the intrinsic nature of the cell's low-pass features. The low-pass property of mouse OHC eM is based on the finding that eM magnitude dissipates within the frequency bandwidth of human speech. In this study, we examined the role of Prestin in sensing broad-range frequencies (4-80 kHz) in mice that use ultrasonic hearing and vocalization (to >100 kHz) for social communication. The audiometric measurements in mice showed that ablation of Prestin did not abolish hearing at frequencies >40 kHz. Acoustic associative behavior tests confirmed that Prestin-knockout mice can learn ultrahigh-frequency sound-coupled tasks, similar to control mice. Ex vivo cochlear Ca²⁺ imaging experiments demonstrated that without Prestin, the OHCs still exhibit ultrahigh-frequency transduction, which in contrast, can be abolished by a universal cation channel blocker, Gadolinium. In vivo salicylate treatment disrupts hearing at frequencies <40 kHz but not ultrahigh-frequency hearing. By pharmacogenetic manipulation, we showed that specific ablation of the OHCs largely abolished hearing at frequencies >40 kHz. These findings demonstrate that cochlear OHCs are the target cells that support ultrahigh-frequency transduction, which does not require Prestin.
Animals ; Cochlea/metabolism* ; Hair Cells, Auditory, Outer/metabolism* ; Hearing ; Humans ; Mammals/metabolism* ; Mice ; Mice, Knockout ; Molecular Motor Proteins/metabolism*