Sheep as a large animal ear model: Middle-ear ossicular velocities and intracochlear sound pressure.

Animals are frequently used for the development and testing of new hearing devices. Dimensions of the middle ear and cochlea differ significantly between humans and commonly used animals, such as rodents or cats. The sheep cochlea is anatomically more like the human cochlea in size and number of turns. This study investigated the middle-ear ossicular velocities and intracochlear sound pressure (ICSP) in sheep temporal bones, with the aim of characterizing the sheep as an experimental model for implantable hearing devices. Measurements were made on fresh sheep temporal bones. Velocity responses of the middle ear ossicles at the umbo, long process of the incus and stapes footplate were measured in the frequency range of 0.25-8 kHz using a laser Doppler vibrometer system. Results were normalized by the corresponding sound pressure level in the external ear canal (PEC). Sequentially, ICSPs at the scala vestibuli and tympani were then recorded with custom MEMS-based hydrophones, while presenting identical acoustic stimuli. The sheep middle ear transmitted most effectively around 4.8 kHz, with a maximum stapes velocity of 0.2 mm/s/Pa. At the same frequency, the ICSP measurements in the scala vestibuli and tympani showed the maximum gain relative to the PEC (24 dB and 5 dB, respectively). The greatest pressure difference across the cochlear partition occurred between 4 and 6 kHz. A comparison between the results of this study and human reference data showed middle-ear resonance and best cochlear sensitivity at higher frequencies in sheep. In summary, sheep can be an appropriate large animal model for research and development of implantable hearing devices.

Retroauricular Approach for Targeted Cochlear Therapy Experiments in Wistar Albino Rats.

BACKGROUND: As the idea of stem cell technology in the treatment of sensorial hearing loss has emerged over the past decades, the need for in vivo models for related experiments has become explicit. One of the most common experimental models for inner ear stem cell delivery experiments is the Wistar albino rat. AIMS: To investigate the surgical anatomy of the temporal bone of the Wistar albino rat with respect to the dissection steps, operative techniques and potential pitfalls of surgery. STUDY DESIGN: Animal experimentation. METHODS: Adult Wistar albino rats were operated on via the retroauricular approach under an operation microscope. The anatomy of the temporal bone, the surgical route to the temporal bulla and the inner ear were investigated. Technical details of surgical steps, complications and potential pitfalls during the surgery were noted. RESULTS: The study group consisted of 40 adult Wistar albino rats. The mean times to reach the bulla and to achieve cochleostomy were 4.3 (2-13 min) and 7.5 min (3.5-22 min), respectively. The mean width of the facial nerve was 0.84 mm (0.42-1.25 mm). The stapedial artery lay nearly perpendicular to the course of the facial nerve (88-93 °C). There were three major complications: two large cochleostomies and one massive bleed from the stapedial artery. CONCLUSION: The facial nerve was the key anatomical landmark in locating the bulla. By retrograde tracing of the facial nerve, it was possible to find the bulla ventral (inferior) to the main trunk. The facial nerve trunk was the upper limit when drilling the bulla. By dissecting the main trunk of the facial nerve and retracting cranially, a large drilling space could be achieved. Our results suggest that the retroauricular approach is an effective, feasible route for inner ear drug delivery experiments in Wistar albino rats.

Relationship Between Electrode-to-Modiolus Distance and Current Levels for Adults With Cochlear Implants.

HYPOTHESIS: Electrode-to-modiolus distance is correlated with clinically programmed stimulation levels. BACKGROUND: Conventional wisdom has long supposed a significant relationship between cochlear implant (CI) stimulation levels and electrode-to-modiolus distance; however, to date, no such formal investigation has been completed. Thus, the purpose of this project was to investigate the relationship between stimulation levels and electrode-to-modiolus distance. A strong correlation between the two would suggest that stimulation levels might be used to estimate electrode-to-modiolus geometry. METHODS: Electrode-to-modiolus distance was determined via CT imaging using validated CI position analysis software in 137 implanted ears from the three manufacturers holding FDA approval in the United States. Analysis included 2,365 total electrodes, with 1,472 from precurved arrays. Distances were compared to clinically programmed C/M levels that were converted to charge units. RESULTS: Mean modiolar distance with perimodiolar and lateral wall electrodes was 0.47 and 1.15 mm, respectively. Mean suprathreshold charge values were significantly different between each manufacturer. When combining all data, we found a moderate positive correlation (r = 0.367, p < 0.01) that was driven both by the different charge values across companies, and that the company with the highest mean charge values only offers straight electrode arrays. When grouped by electrode type, however, we found a weak correlation (r = 0.12, p < 0.01) for perimodiolar array electrodes only. When considering a single array type from any one manufacturer, only one was observed where distance mildly predicted charge. CONCLUSION: Our results suggest that electrode distance minimally contributes to the current level required for suprathreshold stimulation.

[Effects of electroacupuncture on cochlea morphology and expression of aquaporins in guinea pigs with endolymphatic hydrops].

OBJECTIVE: To observe the effects of electroacupuncture (EA) on cochlea morphology and expression of aquaporin 1 (AQP1) in guinea pigs with endolymphatic hydrops, so as to explore the possible mechanism of EA on endolymphatic hydrops. METHODS: Forty guinea pigs were randomly divided into a blank group, a model group, a medication group and an EA group, 10 guinea pigs in each one. Model of endolymphatic hydrops was established by using intraperitoneal injection of aldosterone. Guinea pigs in the blank group and model group were treated with identical immobilization as EA group but no treatment was given; guinea pigs in the medication group were treated with intragastric administration of hydrochlorothiazide at a dose of 5 mg/kg, once a day for consecutive 10 days; guinea pigs in the EA group were treated with' EA at "Baihui" (GV 20) and "Tinggong"(SI 19), once a day for consecutive 10 days. The serum ionic concentration in each group was tested by turbidimetric method; hematoxylin-eosin staining was used to measure the severity of cochlea hydrops; immunohistochemical method was used to observe the expression of AQP1 in the cochlea. RESULTS: (1) There was no endolymphatic hydrops in the blank group, moderate-severe endolymphatic hydrops in the model group and slight endolymphatic hydrops in the EA group and medication group. (2) The concentration of K+ and Ca2+ in the EA group was higher than that in the model group and medication group (all P<0. 01); the concentration of Na+ was lower than that in the model group (P< 0. 01) but higher than that in the medication group (P<0. 01); the concentration of Cl- was higher than that in the medication group (P<0. 01), but not significantly different from the model group (P>0. 05). (3) The ratio of expression area of AQP1 in the model group was lower than that in the blank group (P<0. 01); the ratio of expression area of AQP1 in the EA group was higher than that in the model group (P<0. 01), and lower than that in the medication group without significant difference (P>0. 05). CONCLUSION: EA could relieve the endolymphatic hydrops in guinea pigs; the mechanism is likely to be related with up-regulating the expression of AQP1 in cochlea and ion concentration might be an important factor involved.

Modeling the utility of binaural cues for underwater sound localization.

The binaural cues used by terrestrial animals for sound localization in azimuth may not always suffice for accurate sound localization underwater. The purpose of this research was to examine the theoretical limits of interaural timing and level differences available underwater using computational and physical models. A paired-hydrophone system was used to record sounds transmitted underwater and recordings were analyzed using neural networks calibrated to reflect the auditory capabilities of terrestrial mammals. Estimates of source direction based on temporal differences were most accurate for frequencies between 0.5 and 1.75 kHz, with greater resolution toward the midline (2°), and lower resolution toward the periphery (9°). Level cues also changed systematically with source azimuth, even at lower frequencies than expected from theoretical calculations, suggesting that binaural mechanical coupling (e.g., through bone conduction) might, in principle, facilitate underwater sound localization. Overall, the relatively limited ability of the model to estimate source position using temporal and level difference cues underwater suggests that animals such as whales may use additional cues to accurately localize conspecifics and predators at long distances.

Bone conduction hearing sensitivity in normal-hearing subjects: transcutaneous stimulation at BAHA vs BCI position.

OBJECTIVE: Bone conduction (BC) stimulation closer to the cochlea has previously been shown to give higher cochlear promontory acceleration measured by laser Doppler vibrometry (LDV). This study is investigating whether stimulation closer to the cochlea also gives improved hearing sensitivity. Furthermore, the study compares shifts in hearing sensitivity (BC thresholds) and ear-canal sound pressure (ECSP). DESIGN: BC hearing thresholds and ECSP have been measured for stimulation at two positions: the existing bone-anchored hearing aid (BAHA) position, and a new bone conduction implant (BCI) position that is located closer to the cochlea. STUDY SAMPLE: The measurements were made on 20 normal-hearing subjects. RESULTS: Depending on frequency, the ipsilateral hearing threshold was 3-14 dB better, and the ipsilateral ECSP was 2-12 dB higher for the BCI than for the BAHA position, with no significant differences between threshold and ECSP shifts at group level for most frequencies, and individually only for some subjects. CONCLUSIONS: It was found that both the objective ECSP and the subjective hearing threshold measurements gave similar improvement as previous LDV measurements for stimulation closer to the cochlea. One exception was that the LDV measurements did not show the improved sensitivity for frequencies below 500 Hz found here.

Age-related hearing loss: GABA, nicotinic acetylcholine and NMDA receptor expression changes in spiral ganglion neurons of the mouse.

Age-related hearing loss - presbycusis - is the number one communication disorder and most prevalent neurodegenerative condition of our aged population. Although speech understanding in background noise is quite difficult for those with presbycusis, there are currently no biomedical treatments to prevent, delay or reverse this condition. A better understanding of the cochlear mechanisms underlying presbycusis will help lead to future treatments. Objectives of the present study were to investigate GABAA receptor subunit α1, nicotinic acetylcholine (nACh) receptor subunit ß2, and N-methyl-d-aspartate (NMDA) receptor subunit NR1 mRNA and protein expression changes in spiral ganglion neurons (SGN) of the CBA/CaJ mouse cochlea, that occur in age-related hearing loss, utilizing quantitative immunohistochemistry and semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) techniques. We found that auditory brainstem response (ABR) thresholds shifted over 40dB from 3 to 48kHz in old mice compared to young adults. DPOAE thresholds also shifted over 40dB from 6 to 49kHz in old mice, and their amplitudes were significantly decreased or absent in the same frequency range. SGN density decreased with age in basal, middle and apical turns, and SGN density of the basal turn declined the most. A positive correlation was observed between SGN density and ABR wave 1amplitude. mRNA and protein expression of GABAAR α1 and AChR ß2 decreased with age in SGNs in the old mouse cochlea. mRNA and protein expression of NMDAR NR1 increased with age in SGNs of the old mice. These findings demonstrate that there are functionally-relevant age-related changes of GABAAR, nAChR, NMDAR expression in CBA mouse SGNs reflecting their degeneration, which may be related to functional changes in cochlear synaptic transmission with age, suggesting biological mechanisms for peripheral age-related hearing loss.

Nonlinear time-domain cochlear model for transient stimulation and human otoacoustic emission.

This paper describes the implementation and performance of a nonlinear time-domain model of the cochlea for transient stimulation and human otoacoustic emission generation. The nonlinearity simulates compressive growth of measured basilar-membrane impulse responses. The model accounts for reflection and distortion-source otoacoustic emissions (OAEs) and simulates spontaneous OAEs through manipulation of the middle-ear reflectance. The model was calibrated using human psychoacoustical and otoacoustic tuning parameters. It can be used to investigate time-dependent properties of cochlear mechanics and the generator mechanisms of otoacoustic emissions. Furthermore, the model provides a suitable preprocessor for human auditory perception models where realistic cochlear excitation patterns are desired.

Analysis of the cochlear microphonic to a low-frequency tone embedded in filtered noise.

The cochlear microphonic was recorded in response to a 733 Hz tone embedded in noise that was high-pass filtered at 25 different frequencies. The amplitude of the cochlear microphonic increased as the high-pass cutoff frequency of the noise increased. The amplitude growth for a 60 dB SPL tone was steeper and saturated sooner than that of an 80 dB SPL tone. The growth for both signal levels, however, was not entirely cumulative with plateaus occurring at about 4 and 7 mm from the apex. A phenomenological model of the electrical potential in the cochlea that included a hair cell probability function and spiral geometry of the cochlea could account for both the slope of the growth functions and the plateau regions. This suggests that with high-pass-filtered noise, the cochlear microphonic recorded at the round window comes from the electric field generated at the source directed towards the electrode and not down the longitudinal axis of the cochlea.

Effects of hyperbaric oxygen and dexamethasone on proinflammatory cytokines of rat cochlea in noise-induced hearing loss.

HYPOTHESIS: To investigate effects of dexamethasone and hyperbaric oxygen therapy (HBOT) on proinflammatory cytokines and hearing levels in the noise-exposed cochlea of rats. BACKGROUND: There is an arising concern about negative effects of early initiation of HBOT on hearing in noise-induced hearing loss. Furthermore, effects of HBOT and dexamethasone on cochlear cytokines are not fully elucidated. METHODS: Twenty-six rats were divided into 3 groups: control, noise, and treatment groups. Five rats served as control group. White noise at 115 dB sound pressure level was applied to the noise group of 4 rats for 10 days. This group was assigned to a positive control group as it was equivalent to treatment groups. The treatment group of 17 rats underwent the same noise exposure, and then, they were divided into 3 groups based on treatment protocol: 5 and 6 rats received HBOT at the third hour and 24th hour after the noise, respectively, and 6 rats received dexamethasone. Auditory brain stem response threshold was measured in all groups before being assigned to the groups, after the noise exposure and right before being killed. Cytokine levels at the cochlear soft tissues were measured using enzyme-linked immunoassay. RESULTS: Final thresholds (10 dB and 5 dB nHL-normal hearing level) of HBOT-24th hour and dexamethasone groups were significantly better than that of untreated noise group (22.5 dB nHL) (p < 0.05). There was no significant difference between HBOT-24th hour group (10 dB nHL) and dexamethasone group (5 dB nHL) (p > 0.05). IL-6 and IL-1ß of HBOT-third hour group (2.30 ng/mg and 185.43 pg/mg) were significantly higher than those of the noise group (0.91 ng/mg and 131.40 pg/mg), dexamethasone group (1.19 ng/mg and 112.29 pg/mg) and HBOT-24th hour group (1.34 ng/mg and 106.69 pg/mg) (p < 0.05). There was no significant difference in IL-6 and IL-1ß of HBOT-24th hour group, dexamethasone group, noise group, and control group (p > 0.05). There was no significant difference in TNF-α of the 3 treatment groups, noise group, and control group (p > 0.05). CONCLUSION: The results showed that the most effective method in the treatment of noise-induced hearing loss was early initiation of dexamethasone therapy. There could be negative effects of HBOT on hearing if it is commenced early after the noise (first 3 h). HBOT treatment, which was started at the 24th hour, was found to be an effective method.

Instrumentation for studies of cochlear mechanics: from von Békésy forward.

Georg von Békésy designed the instruments needed for his research. He also created physical models of the cochlea allowing him to manipulate the parameters (such as volume elasticity) that could be involved in controlling traveling waves. This review is about the specific devices that he used to study the motion of the basilar membrane thus allowing the analysis that lead to his Nobel Prize Award. The review moves forward in time mentioning the subsequent use of von Békésy's methods and later technologies important for motion studies of the organ of Corti. Some of the seminal findings and the controversies of cochlear mechanics are mentioned in relation to the technical developments.

Traveling waves on the organ of corti of the chinchilla cochlea: spatial trajectories of inner hair cell depolarization inferred from responses of auditory-nerve fibers.

Spatial magnitude and phase profiles for inner hair cell (IHC) depolarization throughout the chinchilla cochlea were inferred from responses of auditory-nerve fibers (ANFs) to threshold- and moderate-level tones and tone complexes. Firing-rate profiles for frequencies ≤2 kHz are bimodal, with the major peak at the characteristic place and a secondary peak at 3-5 mm from the extreme base. Response-phase trajectories are synchronous with peak outward stapes displacement at the extreme cochlear base and accumulate 1.5 period lags at the characteristic places. High-frequency phase trajectories are very similar to the trajectories of basilar-membrane peak velocity toward scala tympani. Low-frequency phase trajectories undergo a polarity flip in a region, 6.5-9 mm from the cochlear base, where traveling-wave phase velocity attains a local minimum and a local maximum and where the onset latencies of near-threshold impulse responses computed from responses to near-threshold white noise exhibit a local minimum. That region is the same where frequency-threshold tuning curves of ANFs undergo a shape transition. Since depolarization of IHCs presumably indicates the mechanical stimulus to their stereocilia, the present results suggest that distinct low-frequency forward waves of organ of Corti vibration are launched simultaneously at the extreme base of the cochlea and at the 6.5-9 mm transition region, from where antiphasic reflections arise.

Probing cochlear tuning and tonotopy in the tiger using otoacoustic emissions.

Otoacoustic emissions (sound emitted from the ear) allow cochlear function to be probed noninvasively. The emissions evoked by pure tones, known as stimulus-frequency emissions (SFOAEs), have been shown to provide reliable estimates of peripheral frequency tuning in a variety of mammalian and non-mammalian species. Here, we apply the same methodology to explore peripheral auditory function in the largest member of the cat family, the tiger (Panthera tigris). We measured SFOAEs in 9 unique ears of 5 anesthetized tigers. The tigers, housed at the Henry Doorly Zoo (Omaha, NE), were of both sexes and ranged in age from 3 to 10 years. SFOAE phase-gradient delays are significantly longer in tigers--by approximately a factor of two above 2 kHz and even more at lower frequencies--than in domestic cats (Felis catus), a species commonly used in auditory studies. Based on correlations between tuning and delay established in other species, our results imply that cochlear tuning in the tiger is significantly sharper than in domestic cat and appears comparable to that of humans. Furthermore, the SFOAE data indicate that tigers have a larger tonotopic mapping constant (mm/octave) than domestic cats. A larger mapping constant in tiger is consistent both with auditory brainstem response thresholds (that suggest a lower upper frequency limit of hearing for the tiger than domestic cat) and with measurements of basilar-membrane length (about 1.5 times longer in the tiger than domestic cat).

Békésy's contributions to our present understanding of sound conduction to the inner ear.

In our daily lives we hear airborne sounds that travel primarily through the external and middle ear to the cochlear sensory epithelium. We also hear sounds that travel to the cochlea via a second sound-conduction route, bone conduction. This second pathway is excited by vibrations of the head and body that result from substrate vibrations, direct application of vibrational stimuli to the head or body, or vibrations induced by airborne sound. The sensation of bone-conducted sound is affected by the presence of the external and middle ear, but is not completely dependent upon their function. Measurements of the differential sensitivity of patients to airborne sound and direct vibration of the head are part of the routine battery of clinical tests used to separate conductive and sensorineural hearing losses. Georg von Békésy designed a careful set of experiments and pioneered many measurement techniques on human cadaver temporal bones, in physical models, and in human subjects to elucidate the basic mechanisms of air- and bone-conducted sound. Looking back one marvels at the sheer number of experiments he performed on sound conduction, mostly by himself without the aid of students or research associates. Békésy's work had a profound impact on the field of middle-ear mechanics and bone conduction fifty years ago when he received his Nobel Prize. Today many of Békésy's ideas continue to be investigated and extended, some have been supported by new evidence, some have been refuted, while others remain to be tested.

Contributions of von Békésy to psychoacoustics.

This paper reviews the contributions of von Békésy to psychoacoustics, comparing his findings and interpretations to those that have emerged since his work. The areas covered include the perception of pitch for pure tones and complex tones, the effect of frequency on the apparent location of pure tones, estimation of the velocity of the traveling wave on the basilar membrane using judgments of lateralization, and the relative loudness of monaural and diotic sounds. While subsequent research has failed to replicate some of his findings, other findings have stood the test of time. There is no doubt that von Békésy made very substantial contributions to psychoacoustic research.

[Effect of intervention of granules of eliminating phlegm and removing blood stasis in cochlear morphology of diabetic rats].

OBJECTIVE: To study the effectiveness of the granules of eliminating phlegm and removing blood stasis (GEPRB) on glucose metabolism for diabetic rats induced by streptozotocin and that on early diabetic rats cochlear histopathology. METHOD: Intraperitoneally inject streptozotocin disposable of dose of 55 mg x kg(-1), so that a model of diabetic rats is created, and treated by, and simultaneously treated by Duxil as matched groups for positive, then observe the blood glucose of each group, the histopathology of cochlear, and any circumstantial change of their microstructures. RESULT: GEPRB can reduce the cumulating thickness of the basilar membrane and that of the vascular pattern, so as to tighten up the sparsity of the spiral ganglion cells. CONCLUSION: GEPRB is rather effective on hypoglycemic for the diabetic rat model, and therapeutic goal can be achieved by improving the pathological changes of the cochlea damaged. The improvement by GEPRB for microangiopathic change of the cochlea capillary is probably the pathological basis of relieving hearing loss.

Brightness-compensated 3-D optical flow algorithm for monitoring cochlear motion patterns.

A method for three-dimensional motion analysis designed for live cell imaging by fluorescence confocal microscopy is described. The approach is based on optical flow computation and takes into account brightness variations in the image scene that are not due to motion, such as photobleaching or fluorescence variations that may reflect changes in cellular physiology. The 3-D optical flow algorithm allowed almost perfect motion estimation on noise-free artificial sequences, and performed with a relative error of <10% on noisy images typical of real experiments. The method was applied to a series of 3-D confocal image stacks from an in vitro preparation of the guinea pig cochlea. The complex motions caused by slow pressure changes in the cochlear compartments were quantified. At the surface of the hearing organ, the largest motion component was the transverse one (normal to the surface), but significant radial and longitudinal displacements were also present. The outer hair cell displayed larger radial motion at their basolateral membrane than at their apical surface. These movements reflect mechanical interactions between different cellular structures, which may be important for communicating sound-evoked vibrations to the sensory cells. A better understanding of these interactions is important for testing realistic models of cochlear mechanics.

Correlation of early auditory potentials and intracochlear electrode insertion properties: an animal model featuring near real-time monitoring.

OBJECTIVE: The goal of this work was to assess electrophysiologic response changes to acoustic stimuli as an intracochlear electrode impacted cochlear structures in an animal model of hearing preservation cochlear implantation. The ultimate goal is to develop efficient procedures for assessing the status of cochlear physiology for intraoperative use. METHODS: Sixteen gerbils and 18 ears were tested. A rigid electrode was inserted through a basal turn cochleostomy and directed toward the basilar membrane/osseous spiral lamina complex. We recorded acoustically evoked early auditory potentials including cochlear microphonics (CMs) and compound action potentials (CAPs) to a short stimulation sequence consisting of one stimulus frequency and intensity as the electrode was advanced. A microendoscope was used to visualize the electrode insertion progress and to identify the site of electrode impact. After each experiment, the site of intracochlear trauma was confirmed using whole mount preparations. RESULTS: Electrophysiologic changes correlated well with the degree and location of trauma. We observed four distinct patterns. In addition, the endoscope in conjunction with the short recording sequence allowed for the detection of response changes that were reversible when the electrode was retracted. These cases were associated with less than full-thickness damage on histology. CONCLUSION: The short recording sequence to obtain acoustically evoked intracochlear potentials and the microendoscope allowed us to detect various levels of cochlear trauma including minor and reversible damage. Recordings of this type are potentially available using current implant technology. Future improvements in the measurements can be expected to improve the efficiency of the recording paradigm to produce a clinically useful tool.

Animal model of cochlear third window in the scala vestibuli or scala tympani.

HYPOTHESIS: The auditory impact of a cochlear third window differs by its location in the scala vestibuli or scala tympani. BACKGROUND: Pathologic third window has been investigated primarily in the vestibular apparatus of animals and humans. Dehiscence of the superior semicircular canal is the clinical model. METHODS: Fat sand rats (n = 11) have a unique inner-ear anatomy that allows easy surgical access. A window was drilled in the bony labyrinth over the scala vestibuli in 1 group (12 ears) and over the scala tympani in another (7 ears) while preserving the membranous labyrinth. Auditory brain stem responses to high- and low-frequency stimuli delivered by air and bone conduction were recorded before and after the procedure. RESULTS: Scala vestibuli group: preoperative air-conduction thresholds to clicks and tone-bursts averaged 8.3 and 9.6 dB, respectively, and bone-conduction thresholds, 4.6 and 3.3 dB, respectively; after fenestration, air-conduction thresholds averaged 40.4 and 41.8 dB, respectively, and bone-conduction thresholds, -1 and 5.6 dB, respectively. Scala tympani group: preoperative air-conduction thresholds to clicks and tone-bursts averaged 8.6 dB each, and bone-conduction thresholds, 7.9 dB and 7.1 dB, respectively; after fenestration, air-conduction thresholds averaged 11.4 and 9.3 dB, respectively, and bone-conduction thresholds, 9.3 and 4.2 dB, respectively. The changes in air- (p = 0.0001) and bone-conduction (p = 0.04) thresholds were statistically significant only in the scala vestibuli group. CONCLUSION: The presence of a cochlear third window over the scala vestibuli, but not over the scala tympani, causes a significant increase in air-conduction auditory thresholds. These results agree with the theoretic model and clinical findings and contribute to our understanding of vestibular dehiscence.

Calcium imaging of inner ear hair cells within the cochlear epithelium of mice using two-photon microscopy.

Mice are an excellent model for studying mammalian hearing and transgenic mouse models of human hearing, loss are commonly available. However, the mouse cochlea is substantially smaller than other animal models routinely used to study cochlear physiology. This makes study of their hair cells difficult. We develop a novel methodology to optically image calcium within living hair cells left undisturbed within the excised mouse cochlea. Fresh cochleae are harvested, left intact within their otic capsule bone, and fixed in a recording chamber. The bone overlying the cochlear epithelium is opened and Reissner's membrane is incised. A fluorescent calcium indicator is applied to the preparation. A custom-built upright two-photon microscope was used to image the preparation using 3-D scanning. We are able to image about one third of a cochlear turn simultaneously, in either the apical or basal regions. Within one hour of animal sacrifice, we find that outer hair cells demonstrate increased fluorescence compared with surrounding supporting cells. This methodology is then used to visualize hair cell calcium changes during mechanotransduction over a region of the epithelium. Because the epithelium is left within the cochlea, dissection trauma is minimized and artifactual changes in hair cell physiology are expected to be reduced.