Inner hair cell stereocilia displacement in response to focal stimulation of the basilar membrane in the ex vivo gerbil cochlea.

The inner hair cells in the mammalian cochlea transduce mechanical signals to electrical signals that provide input to the auditory nerve. The spatial-temporal displacement of the inner hair cell stereocilia (IHCsc) relative to basilar membrane (BM) displacement is central to characterizing the transduction process. This study specifically focuses on measuring displacement of the stereocilia hair bundles in the radial dimensions where they are most sensitive. To simplify the mechanical response of the cochlear partition, a mechanical probe was used to drive the BM. Optical imaging was used to measure radial displacement of the inner hair cell stereocilia local to the probe in ex vivo gerbil cochleae. The mechanical probe displaced the BM in the transverse direction using sinusoidal stimuli with frequencies ranging from 10 Hz to 42.5 kHz. IHCsc displacement measurements were made in the radial dimension as a function of their longitudinal location along the length of the BM. The results were used to quantify the frequency response, longitudinal space coupling, traveling wave velocity, and wavelength of the radial displacement of the stereocilia. The measurements were centered at two best frequency locations along the BM: Proximal to the round window (first turn), and in the second turn. At both locations, frequency tuning was seen that was consistent with published place maps. At both locations, traveling waves were observed simultaneously propagating basal and apical from the probe. The velocity of the traveling waves at the center frequency (CF) of the location was higher in the first turn than in the second. As the stimulus frequency increased and approached CF for a location, the traveling wavelength decreased. Differential motion of the BM and IHCsc was observed in the second turn as the stimulus frequency increased toward CF. The longitudinal coupling measured in this study was longer than observed in previous studies. In summary the results suggest that the shape of the wave patterns present on the BM are not sufficient to characterize the displacement of the IHCsc.

Significance of the Microfluidic Flow Inside the Organ of Corti.

We study the vibration modes of a short section in the middle turn of the gerbil cochlea including both longitudinal and radial interstitial fluid spaces between the pillar cells (PC) and the sensory hair cells to determine the role of the interstitial fluid flow within the organ of corti (OoC). Three detailed finite element (FE) models of the cochlear short section (CSS) are studied. In model 1, the CSS is without fluids; model 2 includes the OoC fluid, but not the exterior scalae fluids; and model 3 is the CSS with both scalae and OoC fluids. We find that: (1) the fundamental mode shape of models 1 or 3 is similar to the classical basilar membrane (BM) bending mode that includes pivoting of the arch of corti, and hence determines the low frequency vibrational mode shape of the cochlea in the presence of the cochlear wave. (2) The fundamental mode shape of model 2 is characterized by a cross-sectional shape change similar to the passive response of the cochlea. This mode shape includes a tilting motion of the inner hair cell (IHC) region, a fluid motion within the tunnel of corti (ToC) in the radial direction and along the OoC, and a bulging motion of the reticular lamina (RL) above the outer hair cell (OHC). Each of these motions provides a plausible mode of excitation of the sensory hair cells. (3) The higher vibrational modes of model 1 are similar to the electrically evoked response within the OoC and suggests that the higher vibrational modes are responsible for the active response of the cochlea. We also observed that the fluid flow through the OoC interstitial space is significant, and the model comparison suggests that the OoC fluid contributes to the biphasic BM motion seen in electrical stimulation experiments. The effect of fluid viscosity on cilium deflection was assessed by performing a transient analysis to calculate the cilium shearing gain. The gain values are found to be within the range of experimentally measured values reported by Dallos et al. (1996, The Cochlea, Springer-Verlag, New York).

A model and experimental approach to the middle ear transfer function related to hearing in the humpback whale (Megaptera novaeangliae).

At present, there are no direct measures of hearing for any baleen whale (Mysticeti). The most viable alternative to in vivo approaches to simulate the audiogram is through modeling outer, middle, and inner ear functions based on the anatomy and material properties of each component. This paper describes a finite element model of the middle ear for the humpback whale (Megaptera novaeangliae) to calculate the middle ear transfer function (METF) to determine acoustic energy transmission to the cochlea. The model was developed based on high resolution computed tomography imaging and direct anatomical measurements of the middle ear components for this mysticete species. Mechanical properties for the middle ear tissues were determined from experimental measurements and published values. The METF for the humpback whale predicted a better frequency range between approximately 15 Hz and 3 kHz or between 200 Hz and 9 kHz based on two potential stimulation locations. Experimental measures of the ossicular chain, tympanic membrane, and tympanic bone velocities showed frequency response characteristics consistent with the model. The predicted best sensitivity hearing ranges match well with known vocalizations of this species.

Simulated masking of right whale sounds by shipping noise: incorporating a model of the auditory periphery.

Many species of large, mysticete whales are known to produce low-frequency communication sounds. These low-frequency sounds are susceptible to communication masking by shipping noise, which also tends to be low frequency in nature. The size of these species makes behavioral assessment of auditory capabilities in controlled, captive environments nearly impossible, and field-based playback experiments are expensive and necessarily limited in scope. Hence, it is desirable to produce a masking model for these species that can aid in determining the potential effects of shipping and other anthropogenic noises on these protected animals. The aim of this study was to build a model that combines a sophisticated representation of the auditory periphery with a spectrogram-based decision stage to predict masking levels. The output of this model can then be combined with a habitat-appropriate propagation model to calculate the potential effects of noise on communication range. For this study, the model was tested on three common North Atlantic right whale communication sounds, both to demonstrate the method and to probe how shipping noise affects the detection of sounds with varying spectral and temporal characteristics.

Elastic modulus of cetacean auditory ossicles.

In order to model the hearing capabilities of marine mammals (cetaceans), it is necessary to understand the mechanical properties, such as elastic modulus, of the middle ear bones in these species. Biologically realistic models can be used to investigate the biomechanics of hearing in cetaceans, much of which is currently unknown. In the present study, the elastic moduli of the auditory ossicles (malleus, incus, and stapes) of eight species of cetacean, two baleen whales (mysticete) and six toothed whales (odontocete), were measured using nanoindentation. The two groups of mysticete ossicles overall had lower average elastic moduli (35.2 ± 13.3 GPa and 31.6 ± 6.5 GPa) than the groups of odontocete ossicles (53.3 ± 7.2 GPa to 62.3 ± 4.7 GPa). Interior bone generally had a higher modulus than cortical bone by up to 36%. The effects of freezing and formalin-fixation on elastic modulus were also investigated, although samples were few and no clear trend could be discerned. The high elastic modulus of the ossicles and the differences in the elastic moduli between mysticetes and odontocetes are likely specializations in the bone for underwater hearing.

A prediction of the minke whale (Balaenoptera acutorostrata) middle-ear transfer function.

The lack of baleen whale (Cetacea Mysticeti) audiograms impedes the assessment of the impacts of anthropogenic noise on these animals. Estimates of audiograms, which are difficult to obtain behaviorally or electrophysiologically for baleen whales, can be made by simulating the audiogram as a series of components representing the outer, middle, and inner ear (Rosowski, 1991; Ruggero and Temchin, 2002). The middle-ear portion of the system can be represented by the middle-ear transfer function (METF), a measure of the transmission of acoustic energy from the external ear to the cochlea. An anatomically accurate finite element model of the minke whale (Balaenoptera acutorostrata) middle ear was developed to predict the METF for a mysticete species. The elastic moduli of the auditory ossicles were measured by using nanoindentation. Other mechanical properties were estimated from experimental stiffness measurements or from published values. The METF predicted a best frequency range between approximately 30 Hz and 7.5 kHz or between 100 Hz and 25 kHz depending on stimulation location. Parametric analysis found that the most sensitive parameters are the elastic moduli of the glove finger and joints and the Rayleigh damping stiffness coefficient ß. The predicted hearing range matches well with the vocalization range.

Analysis of the cochlear amplifier fluid pump hypothesis.

We use analysis of a realistic three-dimensional finite-element model of the tunnel of Corti (ToC) in the middle turn of the gerbil cochlea tuned to the characteristic frequency (CF) of 4 kHz to show that the anatomical structure of the organ of Corti (OC) is consistent with the hypothesis that the cochlear amplifier functions as a fluid pump. The experimental evidence for the fluid pump is that outer hair cell (OHC) contraction and expansion induce oscillatory flow in the ToC. We show that this oscillatory flow can produce a fluid wave traveling in the ToC and that the outer pillar cells (OPC) do not present a significant barrier to fluid flow into the ToC. The wavelength of the resulting fluid wave launched into the tunnel at the CF is 1.5 mm, which is somewhat longer than the wavelength estimated for the classical traveling wave. This fluid wave propagates at least one wavelength before being significantly attenuated. We also investigated the effect of OPC spacing on fluid flow into the ToC and found that, for physiologically relevant spacing between the OPCs, the impedance estimate is similar to that of the underlying basilar membrane. We conclude that the row of OPCs does not significantly impede fluid exchange between ToC and the space between the row of OPC and the first row of OHC-Dieter's cells complex, and hence does not lead to excessive power loss. The BM displacement resulting from the fluid pumped into the ToC is significant for motion amplification. Our results support the hypothesis that there is an additional source of longitudinal coupling, provided by the ToC, as required in many non-classical models of the cochlear amplifier.

Basilar membrane tension calculations for the gerbil cochlea.

Anatomical studies suggest that the basilar membrane (BM) supports a radial tension, which is potentially important in cochlear mechanics. Assuming that the tension exists, we have calculated its magnitude from measurements of BM stiffness, longitudinal coupling, and geometry using a BM model. Results for the gerbil cochlea show that the tension decreases from the base to the apex of the cochlea and generates a tensile stress that is comparable in magnitude to the stress generated in other physiological systems. The model calculations are augmented by experiments that investigate the source of BM tension. The experimental results suggest that BM tension is maintained by the spiral ligament.

Evidence for outer hair cell driven oscillatory fluid flow in the tunnel of corti.

Outer hair cell (OHC) somatic motility plays a key role in mammalian cochlear frequency selectivity and hearing sensitivity, but the mechanism of cochlear amplification is not well understood and remains a matter of controversy. We have visualized and quantified the effects of electrically evoked OHC somatic motility within the gerbil organ of Corti using an excised cochlear preparation. We found that OHC motility induces oscillatory motion of the medial olivocochlear fibers where they cross the tunnel of Corti (ToC) in their course to innervate the OHCs. We show that this motion is present at physiologically relevant frequencies and remains at locations distal to the OHC excitation point. We interpret this fiber motion to be the result of oscillatory fluid flow in the ToC. We show, using a simple one-dimensional hydromechanical model of the ToC, that a fluid wave within the tunnel can travel without significant attenuation for distances larger than the wavelength of the cochlear traveling wave at its peak. This ToC fluid wave could interact with the cochlear traveling wave to amplify the motion of the basilar membrane. The ToC wave could also provide longitudinal coupling between adjacent sections of the basilar membrane, and such coupling may be critical for cochlear amplification.

Imaging electrically evoked micromechanical motion within the organ of corti of the excised gerbil cochlea.

The outer hair cell (OHC) of the mammalian inner ear exhibits an unusual form of somatic motility that can follow membrane-potential changes at acoustic frequencies. The cellular forces that produce this motility are believed to amplify the motion of the cochlear partition, thereby playing a key role in increasing hearing sensitivity. To better understand the role of OHC somatic motility in cochlear micromechanics, we developed an excised cochlea preparation to visualize simultaneously the electrically-evoked motion of hundreds of cells within the organ of Corti (OC). The motion was captured using stroboscopic video microscopy and quantified using cross-correlation techniques. The OC motion at approximately 2-6 octaves below the characteristic frequency of the region was complex: OHC, Deiter's cell, and Hensen's cell motion were hundreds of times larger than the tectorial membrane, reticular lamina (RL), and pillar cell motion; the inner rows of OHCs moved antiphasic to the outer row; OHCs pivoted about the RL; and Hensen's cells followed the motion of the outer row of OHCs. Our results suggest that the effective stimulus to the inner hair cell hair bundles results not from a simple OC lever action, as assumed by classical models, but by a complex internal motion coupled to the RL.

A hydromechanical biomimetic cochlea: experiments and models.

The construction, measurement, and modeling of an artificial cochlea (ACochlea) are presented in this paper. An artificial basilar membrane (ABM) was made by depositing discrete Cu beams on a piezomembrane substrate. Rather than two fluid channels, as in the mammalian cochlea, a single fluid channel was implemented on one side of the ABM, facilitating the use of a laser to detect the ABM vibration on the other side. Measurements were performed on both the ABM and the ACochlea. The measurement results on the ABM show that the longitudinal coupling on the ABM is very strong. Reduced longitudinal coupling was achieved by cutting the membrane between adjacent beams using a laser. The measured results from the ACochlea with a laser-cut ABM demonstrate cochlear-like features, including traveling waves, sharp high-frequency rolloffs, and place-specific frequency selectivity. Companion computational models of the mechanical devices were formulated and implemented using a circuit simulator. Experimental data were compared with simulation results. The simulation results from the computational models of the ABM and the ACochlea are similar to their experimental counterparts.

Progression of hair cell ejection and molecular markers of apoptosis in the avian cochlea following gentamicin treatment.

Aminoglycoside treatment induces caspase-dependent apoptotic death in inner ear sensory hair cells. The timing of apoptotic signaling in sensory hair cells following systemic aminoglycoside treatment has not been characterized in vivo. We administered a single subcutaneous injection of the aminoglycoside gentamicin (300 mg/kg) to 12-16-day-old chicks and used immunocytochemical techniques to document the following responses in affected hair cells: T-cell restricted intracellular antigen-related protein (TIAR) translocation from the nucleus to the cytoplasm, cytochrome c release from the mitochondria, caspase-3 activation, nuclear condensation, and an orderly progression of hair cell ejection from the proximal end of the basilar papilla. Hair cells in the proximal tip exhibited TIAR translocation from the nucleus and aggregation into punctate granules in the cytoplasm 12 hours after injection and the response progressed distally. Cytochrome c release from the mitochondria into the cytoplasm and caspase-3 activation were observed in affected hair cells immediately prior to and during ejection. Hair cell ejection occurred between 30 and 54 hours after injection, beginning in the proximal tip and progressing distally. Nuclear condensation accompanied ejection while the loss of: 1) membrane integrity; 2) phalloidin labeling of F-actin; and 3) TO-PRO-1 labeling of nuclear contents occurred within 48 hours following ejection. Our results present a timeline of aminoglycoside-induced inner ear sensory hair cell apoptotic death that includes an 18-hour window between the initial apoptotic response and the later stages of programmed death signaling that accompany ejection and a gradual breakdown of hair cells following ejection.