BACKGROUND: Cervical vestibular evoked myogenic potential (cVEMP) testing is a strong tool that enables objective determination of balance functions in humans. However, it remains unknown whether cVEMP correctly expresses vestibular disorder in mice. OBJECTIVE: In this study, correlations of cVEMP with scores for balance-related behavior tests including rotarod, beam, and air-righting reflex tests were determined in ICR mice with vestibular disorder induced by 3,3'-iminodipropiontrile (IDPN) as a mouse model of vestibular disorder. METHODS: Male ICR mice at 4 weeks of age were orally administered IDPN in saline (28 mmol/kg body weight) once. Rotarod, beam crossing, and air-righting reflex tests were performed before and 3-4 days after oral exposure one time to IDPN to determine balance functions. The saccule and utricles were labeled with fluorescein phalloidin. cVEMP measurements were performed for mice in the control and IDPN groups. Finally, the correlations between the scores of behavior tests and the amplitude or latency of cVEMP were determined with Spearman's rank correlation coefficient. Two-tailed Student's t test and Welch's t test were used to determine a significant difference between the two groups. A difference with p < 0.05 was considered to indicate statistical significance. RESULTS: After oral administration of IDPN at 28 mmol/kg, scores of the rotarod, beam, and air-righting reflex tests in the IDPN group were significantly lower than those in the control group. The numbers of hair cells in the saccule, utricle, and cupula were decreased in the IDPN group. cVEMP in the IDPN group was significantly decreased in amplitude and increased in latency compared to those in the control group. cVEMP amplitude had significant correlations with the numbers of hair cells as well as scores for all of the behavior tests in mice. CONCLUSIONS This study demonstrated impaired cVEMP and correlations of cVEMP with imbalance determined by behavior tests in a mouse model of vestibular disorder.
Animals ; Behavior, Animal ; drug effects ; physiology ; Disease Models, Animal ; Hair Cells, Vestibular ; pathology ; Male ; Mice ; Mice, Inbred ICR ; Nitriles ; adverse effects ; Postural Balance ; drug effects ; physiology ; Saccule and Utricle ; pathology ; Sensation Disorders ; chemically induced ; physiopathology ; Vestibular Diseases ; chemically induced ; diagnosis ; pathology ; physiopathology ; Vestibular Evoked Myogenic Potentials ; drug effects ; physiology ; Vestibular Function Tests

Hypotension is one of the potential causes of dizziness. In this review, we summarize the studies published in recent years about the electrophysiological and pharmacological mechanisms of hypotension-induced dizziness and the role of the vestibular system in the control of blood pressure in response to hypotension. It is postulated that ischemic excitation of the peripheral vestibular hair cells as a result of a reduction in blood flow to the inner ear following hypotension leads to excitation of the central vestibular nuclei, which in turn may produce dizziness after hypotension. In addition, excitation of the vestibular nuclei following hypotension elicits the vestibulosympathetic reflex, and the reflex then regulates blood pressure by a dual-control (neurogenic and humoral control) mechanism. In fact, recent studies have shown that peripheral vestibular receptors play a role in the control of blood pressure through neural reflex pathways. This review illustrates the dual-control mechanism of peripheral vestibular receptors in the regulation of blood pressure following hypotension.
Blood Pressure* ; Dizziness ; Ear, Inner ; Epinephrine ; Glutamic Acid ; Hair Cells, Vestibular ; Hypotension* ; Reflex ; Vestibular Nuclei

The present study was aimed to investigate the effect of hydrogen peroxide (H₂O₂, oxygen free radical donator) on the current of large-conductance calcium-activated potassium channels (BK(Ca) channels) in isolated outer hair cells of old guinea pig cochlea, and to explore the underlying mechanism. Outer hair cells of old guinea pig cochlea were acutely enzyme-isolated, and currents were recorded by whole-cell patch clamp. The results showed that, rapid activation and non-deactivation electric currents with a string of large amplitude were recorded. Activation voltage of the current was above -40 - -30 mV. The amplitude of current was increased continuously with the rising of membrane potential. The current showed characteristics of outward rectification without "rundown" phenomenon. IbTX (100 nmol/L) could completely block the activity of channel, which confirmed BK(Ca) channel's current. BK(Ca) current amplitude and peak current density increased with the increment of H₂O₂ concentration (1, 2, 4 μmol/L), showing concentration-dependent activation by H₂O₂. Our results suggest that oxygen free radical/BK(Ca) pathway may be able to adjust the balance of intracellular calcium in outer hair cells.
Animals ; Calcium ; metabolism ; Cochlea ; cytology ; Guinea Pigs ; Hair Cells, Vestibular ; drug effects ; Hydrogen Peroxide ; pharmacology ; Large-Conductance Calcium-Activated Potassium Channels ; metabolism ; Membrane Potentials

BACKGROUND AND OBJECTIVES: Gentamicin (GM) is well known for its vestibulotoxicity. There have been many reports about vestibulotoxicity, however, its mechanism is still unclear. So far, it is known that GM affects the voltage-dependent K+ current and nitric oxide (NO) production. Epigallocatechin-3-gallate (EGCG) is the major component of green tea and is known to have anti-oxidative and anti-toxic effect. This study was undertaken to investigate the protective effect of EGCG against gentamicin on vestibular hair cell (VHC). MATERIALS AND METHOD: White guinea pigs (200-250 g) were rapidly decapitated and the temporal bones were immediately removed. Under a dissecting microscope, the crista ampullaris was obtained. The dissociated VHCs were transferred into a recording chamber mounted onto an inverted microscope. Whole-cell membrane currents and potentials were recorded using standard patch-clamp techniques. In addition, measurements of NO production were obtained using the NO-sensitive dye, 4,5-diamino-fluorescein diacetate (DAF-2DA). RESULTS: Type I VHCs Voltage-dependent K+ current was activated from low depolarizing stimulation. As the stimulation increased, higher current was detected. Voltage-dependent K+ current in type I VHCs was decreased when GM (200 microM) was administrated and GM effects of K+ current inhibition was significantly blocked by EGCG. Extracellular GM-induced an increase in DAF-2DA fluorescence, which thus indicates NO production in VHCs. Also, the GMinduced NO production was inhibited by EGCG. CONCLUSION: GM inhibits voltage-dependent K+ current by releasing NO in isolated type I VHCs. EGCG blocks this inhibitory effects, suggesting a protective role on GM vestibulotoxicity.
Animals ; Fluorescence ; Gentamicins* ; Guinea Pigs* ; Hair Cells, Vestibular* ; Membranes ; Nitric Oxide ; Patch-Clamp Techniques ; Semicircular Ducts ; Tea ; Temporal Bone

Bilateral loss of vestibular sensation from injuries of vestibular hair cells causes individuals suffering poor vision during head movement, postural instability, chronic disequilibrium, and cognitive distraction. A vestibular prosthesis analogous to cochlear implants but designed to modulate vestibular nerve activity during head movement should improve quality of life for these chronically dizzy individuals. An implantable prosthesis that partly restores normal activity on branches of the vestibular nerve should improve quality of life for individuals disabled by this disorder. There have been many efforts to develop and restore 3-dimensional angular vestibule-ocular reflex and the Johns Hopkins vestibular neuro-engineering laboratory has been developing a head-mounted multichannel vestibular prosthesis that restores sufficient semicircular canal function to partially recreate a normal 3-dimensional angular vestibulo-ocular reflex. In this review, their results are described.
Cochlear Implants ; Dizziness ; Ear, Inner ; Hair Cells, Vestibular ; Head Movements ; Prostheses and Implants ; Quality of Life ; Reflex ; Reflex, Vestibulo-Ocular ; Semicircular Canals ; Sensation ; Stress, Psychological ; Vestibular Nerve ; Vision, Ocular

OBJECTIVE: To explore the function of outer hair cells and its influencing factors in the workers of occupational exposure to low frequency electric and magnetic fields. METHOD: 502 high-strength LF EMFs-exposed workers were taken as the study subjects. The field intensity indicator and noise analyzer were employed for the examination of the electromagnetic energy intensity and noise value at the working sites. Self-administered questionnaires were adopted. Universal hearing screening by distortion product otoacoustic emission (DPOAE) and pure tone audiometry (PTA) were done. 100 workers who had abnormal POAE were taken as the observation group and 100 workers who had normal DPOAE were taken as the control group. The workers need be confirmed with ABR testing when the pure tone threshold of two consecutive frequencies were above 20 dB HL or DPOAE detection in any frequency did not pass. Multiple factor analysis with logistic regression was performed for the risk factors. RESULT: The average electric power density in job locations was 21-38 KV/m, which was higher than national standard (< 5 kv/m). Average noise-level in job locations was 52-65 dBHL, which was within the standard (< 85 dB(A). Questionnaire presented that low frequency electric and magnetic fields might increase the incidence of headache, insomnia and tinnitus in the observed group. The incidence of abnormal DPOAE was higher in the subjects of the observed group (P < 0.01). At 676, 933, 3616, 5 130, 7253 Hz, the DPOAE amplitudes of the observed group workers were significantly lower than the control group. Multivariate analysis showed that the risk factors of abnormal DPOAE included exposure of EMF, length of service, daily exposure time, conservatory measures might be protective factors. CONCLUSION The changes of DPOAE indicated that the exposure to low frequency electric and magnetic fields had a subtle, discreet and localized impairing effects on outer hair cells. Exposure of low frequency electric and magnetic fields, length of service, daily exposure time might be risk factors, conservatory measures might be protective factors.
Auditory Threshold ; Case-Control Studies ; Electromagnetic Fields ; adverse effects ; Hair Cells, Vestibular ; pathology ; Humans ; Magnetic Fields ; adverse effects ; Male ; Noise ; adverse effects ; Occupational Exposure ; Surveys and Questionnaires

OBJECTIVETo investigate the mechanism of intense noise-induced apoptosis of vestibular hair cells in guinea pigs and the effect of phosphorylated c-Jun N-terminal kinase (JNK) signal transduction pathway in intense noise-induced apoptosis of vestibular hair cells.

METHODSThirty-two guinea pigs were randomly and equally divided into 1, 5, and 15 d experimental groups and control group. The guinea pigs in the experimental groups were exposed to 4 kHz narrow-band noise at 120 dB SPL for 4 h and then subjected to measurement of auditory brainstem response at 1, 5, or 15 d after noise exposure. In each group, four guinea pigs were used to prepare paraffin sections of vestibular hair cells, and the rest for extraction of total protein from vestibular hair cells. The apoptosis of vestibular hair cells was detected by terminal deoxynucleotidyl transferase (TdT)-mediated d-UTP nick-end labeling (TUNEL). The expression levels of p-JNK and pc-Jun were measured by immunohistochemistry and Western blot.

RESULTSTUNEL-positive cells were found in the vestibular hair cells in the experimental groups, most in the 1 d experimental group and least in the 15 d experimental group, but no positive cells were found in the control group. The immunohistochemical results showed that p-JNK and pc-Jun were detected in the cell nuclei in the experimental groups, but no p-JNK- and pc-Jun-positive cells were found in the control group. The Western blot showed that p-JNK and pc-Jun were increased and activated quickly at 1d after noise exposure, reached the peak levels at 5 d after noise exposure, and were then decreased gradually, but they were still at relatively high levels at 15 d after noise exposure.

CONCLUSIONIntense noise can cause injury to vestibular hair cells by inducing cell apoptosis, and p-JNK marks the activation of JNK signal transduction pathway, suggesting that JNK signal transduction pathway plays an important role in intense noise-induced apoptosis of vestibular hair cells in guinea pigs.

Animals ; Apoptosis ; Guinea Pigs ; Hair Cells, Vestibular ; cytology ; MAP Kinase Kinase 4 ; metabolism ; Noise ; adverse effects ; Phosphorylation ; Signal Transduction

OBJECTIVES: Aminoglycosides are commonly used antibiotic agents, and they are known to generate free oxygen radicals within the inner ear and to cause vestibulo-cochlear toxicity and permanent damage to the sensory hair cells and neurons. Melatonin, a pineal secretory product, has the properties of being a powerful direct and indirect antioxidant. The aim of the present study was to prove the antioxidant effect of melatonin against gentamicin-induced ototoxicty. METHODS: The utricular maculae of Sprague-Dawley rats were prepared from postnatal day 2-4, and these maculae were were divided into 6 groups as follows: 1) control, 2) melatonin only, 3) gentamicin only, and 4), 5), and 6) gentamicin plus melatonin (10, 50, and 100 micrometer, respectively). To count the number of hair cells, 5 utricles from each group were stained with phalloidin-FITC on the 1st, 4th, and 7th days after drug administration. Reactive oxygen species (ROS) was assessed by using the fluorescent probe hydrofluorescent diacetate acetyl ester. The caspase-3 activity was also examined with using the fluorescent caspase-3 substrate and performing Western blotting. RESULTS: The result of this study showed that gentamicin induced the loss of utricular hair cells, and this loss of hair cells was significantly attenuated by co-administration of melatonin. Melatonin reduced ROS production and caspase-3 activation in the gentamicin treated utricular hair cells. CONCLUSION: Our findings conclusively reveal that melatonin has protective effects against gentamicin-induced hair cell loss in the utricles of rat by inhibiting both ROS production and caspase-3 activity.
Aminoglycosides ; Animals ; Antioxidants ; Blotting, Western ; Caspase 3 ; Ear, Inner ; Gentamicins ; Hair ; Hair Cells, Vestibular ; Melatonin ; Neurons ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; Saccule and Utricle

BACKGROUND AND OBJECTIVES: The purpose of this study was to investigate the effects of low level laser for the prevention and treatment of aminoglycoside-induced vestibular ototoxicity. MATERIALS AND METHOD: An organotypic culture of 2 to 4 days old rat utricular maculae was established. Rats were divided into 6 groups according to the treadtment of the utricles: G (gentamicintreated), L (laser-irradiated), LG (laser-irradiated and gentamicin-treated), GL (gentamicin-treated and laser-irradiated), LGL (gentamicin-treated during laser-irradiated) and C (control). After organotypic culture, the utricles of 6 groups were examined by confocal laser scanning electron microscope and scanning electron microscope. The results of each group were compared with each other by statistical methods. RESULTS: The number of vestibular hair cells of the group G was smaller compared to that of the group C. The group L had no difference compared with the group C. The groups LG and GL showed more vestibular hair cells compared with the group G. The group LG showed more vestibular hair cells than the group GL. The group LGL showed most vestibular hair cells compared to that of the groups G, LG, and GL. CONCLUSION: The most effective treatment of aminoglycosideinduced vestibular otoxicity is the irradiation of low level laser before and after the insult of the aminoglycoside. Further clinical studies using low level laser were needed to prevent aminoglycoside-induced ototoxicity and to promote the regeneration of vestibular hair cells.
Animals ; Electrons ; Gentamicins ; Hair Cells, Vestibular ; Rats ; Regeneration ; Saccule and Utricle

BACKGROUND AND OBJECTIVES: Voltage dependent calcium channel (VDCC) mediates calcium ion influx and controls neurotransmitter release in excitable cells. Hair cells in vertebrates cochlea are known to express L-type VDCC. The purpose of this study was to measure calcium current from hair cells to investigate basic activity and characteristics of VDCC. MATERIALS AND METHOD: We measured calcium current in hair cells of the chicken's auditory organ, the basilar papilla analogous to the mammalian cochlea, in whose L-type, dihydropyridinesensitive calcium channels predominate and in vestibular hair cells from cristae. Calcium currentthrough VDCC was isolated in voltage-clamp recording using Cesium, Tetraethylammonium, 4- aminopyridine and apamin to block the much larger potassium currents. Various concentrations of internal calcium buffer, ethylene glycol tetraacetic acid (EGTA) or 1,2-bis (o-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid (BAPTA) were used. RESULTS: The higher the buffer concentration, the larger the current size were ; they were significantly larger in 10 mM of calcium buffer concentration (ANOVA, p< 0.05). There was no difference in calcium current between cochlear and vestibular hair cells. CONCLUSION: We could successfully isolate stable inward calcium current from chick hair cells. This experiment can be used as a basic method to understand neurotransmission process between hair cells and afferent neurons.
Apamin ; Calcium ; Calcium Channels ; Calcium Channels, L-Type ; Cesium ; Cochlea ; Egtazic Acid ; Ethylenes ; Hair ; Hair Cells, Vestibular ; Neurons, Afferent ; Neurotransmitter Agents ; Organ of Corti ; Potassium ; Synaptic Transmission ; Tetraethylammonium ; Vertebrates