Basilar Membrane and Tectorial Membrane Stiffness in the CBA/CaJ Mouse

The mouse has become an important animal model in understanding cochlear function. Structures, such as the tectorial membrane or hair cells, have been changed by gene manipulation, and the resulting effect on cochlear function has been studied. To contrast those findings, physical properties of the basilar membrane (BM) and tectorial membrane (TM) in mice without gene mutation are of great importance. Using the hemicochlea of CBA/CaJ mice, we have demonstrated that tectorial membrane (TM) and basilar membrane (BM) revealed a stiffness gradient along the cochlea. While a simple spring mass resonator predicts the change in the characteristic frequency of the BM, the spring mass model does not predict the frequency change along the TM. Plateau stiffness values of the TM were 0.6 ± 0.5, 0.2 ± 0.1, and 0.09 ± 0.09 N/m for the basal, middle, and upper turns, respectively. The BM plateau stiffness values were 3.7 ± 2.2, 1.2 ± 1.2, and 0.5 ± 0.5 N/m for the basal, middle, and upper turns, respectively. Estimations of the TM Young’s modulus (in kPa) revealed 24.3 ± 25.2 for the basal turns, 5.1 ± 4.5 for the middle turns, and 1.9 ± 1.6 for the apical turns. Young’s modulus determined at the BM pectinate zone was 76.8 ± 72, 23.9 ± 30.6, and 9.4 ± 6.2 kPa for the basal, middle, and apical turns, respectively. The reported stiffness values of the CBA/CaJ mouse TM and BM provide basic data for the physical properties of its organ of Corti.     

The Spatial Buildup of Compression and Suppression in the Mammalian Cochlea

We recorded responses of the gerbil basilar membrane (BM) to wideband tone complexes. The intensity of one component was varied and the effects on the amplitude and phase of the others were assessed. This suppression paradigm enabled us to vary probe frequency and suppressor frequency independently, allowing the use of simple scaling arguments to analyze the spatial buildup of the nonlinear interaction between traveling waves. Most suppressors had the same effects on probe amplitude and phase as did wideband intensity increments. The main exception were suppressors above the characteristic frequency (CF) of the recording location, for which the frequency range of most affected probes was not constant, but shifted upward with suppressor frequency. BM displacement reliably predicted the effectiveness of low-side suppressors, but not high-side suppressors. We found “anti-suppression” of probes well below CF, i.e., suppressor-induced enhancement of probe response amplitude. Large (>1 cycle) phase effects occurred for above-CF probes. Phase shifts varied nonmonotonically, but systematically, with suppressor level, probe frequency, and suppressor frequency, reconciling apparent discrepancies in the literature. The analysis of spatial buildup revealed an accumulation of local effects on the propagation of the traveling wave, with larger BM displacement reducing the local forward gain. The propagation speed of the wave was also affected. With larger BM displacement, the basal portion of the wave slowed down, while the apical part sped up. This framework of spatial buildup of local effects unifies the widely different effects of overall intensity, low-side suppressors, and high-side suppressors on BM responses.     

Energy Flux in the Cochlea: Evidence Against Power Amplification of the Traveling Wave

Traveling waves in the inner ear exhibit an amplitude peak that shifts with frequency. The peaking is commonly believed to rely on motile processes that amplify the wave by inserting energy. We recorded the vibrations at adjacent positions on the basilar membrane in sensitive gerbil cochleae and tested the putative power amplification in two ways. First, we determined the energy flux of the traveling wave at its peak and compared it to the acoustic power entering the ear, thereby obtaining the net cochlear power gain. For soft sounds, the energy flux at the peak was 1 ± 0.6 dB less than the middle ear input power. For more intense sounds, increasingly smaller fractions of the acoustic power actually reached the peak region. Thus, we found no net power amplification of soft sounds and a strong net attenuation of intense sounds. Second, we analyzed local wave propagation on the basilar membrane. We found that the waves slowed down abruptly when approaching their peak, causing an energy densification that quantitatively matched the amplitude peaking, similar to the growth of sea waves approaching the beach. Thus, we found no local power amplification of soft sounds and strong local attenuation of intense sounds. The most parsimonious interpretation of these findings is that cochlear sensitivity is not realized by amplifying acoustic energy, but by spatially focusing it, and that dynamic compression is realized by adjusting the amount of dissipation to sound intensity.     

Point-impedance characterization of the basilar membrane in a three-dimensional cochlea model

In this paper we derive which impedance definition appropriately represents the basilar membrane in a simple three-dimensional model (de Boer's 'block model') of the cochlea. The starting point of our considerations is a system of parallel visco-elastic beams as characterization of the basilar membrane. It is possible to transform this representation into an impedance concept by observing that the membrane velocity is to a very good approximation distributed over the width as a centred half cosine function, independent of the pressure distribution. The ensuing impedance definition is more accurate than the one proposed by de Boer (Hearing Res. 4, 53-77, 1981). The improvement to the model solutions is moderate, however, as appears from numerical calculations of the basilar membrane velocity.     

Basilar membrane tuning properties in the specialised cochlea of the CF-bat, Rhinolophus ferrumequinum

The greater horseshoe bat has greatly expanded frequency mapping, and morphological specialisations, in the first half turn of its cochlea and a sudden transition to normal mapping. Amplitude and phase of vibration have been measured on various structures in the expanded and normal regions and have not revealed any sharply tuned responses. Amplitudes are much lower than those found in other species and frequently show a deep notch in the 77–84 kHz region. The high-frequency cut-off frequencies are tonotopically organised but deviate from the Bruns map, so that hair-cell tuning appears to occur at a frequency at which basilar membrane vibration is small. In the basal region, phase differences were frequently found between the inner and outer parts of the basilar membrane. These appear to be due to interaction between two components of motion and are probably not indicative of a further filtering mechanism. There is no evidence for reflection of the travelling wave at the transition.     

Response Characteristics in the Apex of the Gerbil Cochlea Studied Through Auditory Nerve Recordings

In this study, we analyze the processing of low-frequency sounds in the cochlear apex through responses of auditory nerve fibers (ANFs) that innervate the apex. Single tones and irregularly spaced tone complexes were used to evoke ANF responses in Mongolian gerbil. The spike arrival times were analyzed in terms of phase locking, peripheral frequency selectivity, group delays, and the nonlinear effects of sound pressure level (SPL). Phase locking to single tones was similar to that in cat. Vector strength was maximal for stimulus frequencies around 500 Hz, decreased above 1 kHz, and became insignificant above 4 to 5 kHz. We used the responses to tone complexes to determine amplitude and phase curves of ANFs having a characteristic frequency (CF) below 5 kHz. With increasing CF, amplitude curves gradually changed from broadly tuned and asymmetric with a steep low-frequency flank to more sharply tuned and asymmetric with a steep high-frequency flank. Over the same CF range, phase curves gradually changed from a concave-upward shape to a concave-downward shape. Phase curves consisted of two or three approximately straight segments. Group delay was analyzed separately for these segments. Generally, the largest group delay was observed near CF. With increasing SPL, most amplitude curves broadened, sometimes accompanied by a downward shift of best frequency, and group delay changed along the entire range of stimulus frequencies. We observed considerable across-ANF variation in the effects of SPL on both amplitude and phase. Overall, our data suggest that mechanical responses in the apex of the cochlea are considerably nonlinear and that these nonlinearities are of a different character than those known from the base of the cochlea.     

Development of the Gerbil Inner Ear Observed in the Hemicochlea

A frequency-dependent change in hearing sensitivity occurs during maturation in the basal gerbil cochlea. This change takes place during the first week after the onset of hearing. It has been argued that the mass of a given cochlear segment decreases during development and thus increases the best frequency. Changes in mass during cochlear maturation have been estimated previously by measuring the changes in cochlear dimensions. Fixed, dehydrated, embedded, or sputter-coated tissues were used in such work. However, dehydration of the tissue, a part of most histological techniques, results in severe distortion of some aspects of cochlear morphology. The present experiments, using a novel preparation, the hemicochlea, show that hydrated structures, such as the tectorial membrane and the basilar membrane hyaline matrix, are up to 100% larger than estimated previous studies. Therefore, the hemicochlea was used to study the development of cochlear morphology in the gerbil between the day of birth and postnatal day 19. We used no protocols that would have resulted in severe distortion of cochlear elements. Consequently, a detailed study of cochlear morphology yields several measures that differ from previously published data. Our experiments confirm growth patterns of the cochlea that include a period of remarkably rapid change between postnatal day 6 and 8. The accelerated growth starts in the middle of the cochlea and progresses toward the base and the apex. In particular, the increase in height of Deiters' cells dominated the change, "pushing" the tectorial membrane toward scala vestibuli. This resulted in a shape change of the tectorial membrane and the organ of Corti. The tectorial membrane was properly extended above the outer hair cells by postnatal day 12. This time coincides with the onset of hearing. The basilar membrane hyaline matrix increased in thickness, whereas the multilayered tympanic cover layer cells decreased to a single band of cells by postnatal day 19. Before and after the period of rapid growth, the observed gross morphological changes are rather small. It is unlikely that dimensional changes of cochlear structures between postnatal days 12 and 19 contribute significantly in the remapping of the frequency-place code in the base of the cochlea. Instead, structural changes affecting the stiffness of the cochlear partition might be responsible for the shift in best frequency.     

Structure of the barn owl's (Tyto alba) inner ear

The basilar papilla of the Barn Owl's (Tyto alba) cochlea was found to be 9.5-11.5 mm long. Histological examination revealed that the sensory hair cells had a characteristic distribution: The proximal half contained mostly typical short cells; tall hair cells were present only on the distal half along with many short cells. Lenticular short cells occupied the proximal tip of the papilla. Another unusual feature of the proximal part was a dense fibrous mass in the basilar membrane. This was absent from the distal one-fourth.     

Phase-Locked Responses to Tones of Chinchilla Auditory Nerve Fibers: Implications for Apical Cochlear Mechanics

Responses to tones with frequency ≤ 5 kHz were recorded from auditory nerve fibers (ANFs) of anesthetized chinchillas. With increasing stimulus level, discharge rate–frequency functions shift toward higher and lower frequencies, respectively, for ANFs with characteristic frequencies (CFs) lower and higher than ∼0.9 kHz. With increasing frequency separation from CF, rate–level functions are less steep and/or saturate at lower rates than at CF, indicating a CF-specific nonlinearity. The strength of phase locking has lower high-frequency cutoffs for CFs >4 kHz than for CFs < 3 kHz. Phase–frequency functions of ANFs with CFs lower and higher than ∼0.9 kHz have inflections, respectively, at frequencies higher and lower than CF. For CFs >2 kHz, the inflections coincide with the tip-tail transitions of threshold tuning curves. ANF responses to CF tones exhibit cumulative phase lags of 1.5 periods for CFs 0.7–3 kHz and lesser amounts for lower CFs. With increases of stimulus level, responses increasingly lag (lead) lower-level responses at frequencies lower (higher) than CF, so that group delays are maximal at, or slightly above, CF. The CF-specific magnitude and phase nonlinearities of ANFs with CFs < 2.5 kHz span their entire response bandwidths. Several properties of ANFs undergo sharp transitions in the cochlear region with CFs 2–5 kHz. Overall, the responses of chinchilla ANFs resemble those in other mammalian species but contrast with available measurements of apical cochlear vibrations in chinchilla, implying that either the latter are flawed or that a nonlinear “second filter” is interposed between vibrations and ANF excitation.     

Stiffness of sensory-cell hair bundles in the isolated guinea pig cochlea

Stiffness of hair bundles on cochlear hair cells was measured in turns 2, 3 and 4 of isolated preparations of the guinea-pig organ of Corti maintained in tissue culture medium. Defined as the force required to produce a linear 1.0 micron deflection of the hair-bundle tip, stiffness is greater for deflection in the excitatory than in the inhibitory direction. The excitatory-to-inhibitory ratio for inner hair cells (IHC) is significantly lower than the ratio for outer hair cells (OHC). Hair-bundle stiffness decreases radially from the first to third rows of OHC. Over the measurement range of 9.0-18.0 mm from the stapes hair-bundle stiffness decreases much more for OHC (88-97%) than for IHC (50%). Although an increase in hair-bundle length with distance from the stapes accounts for some of the observed stiffness decrease, the major decrease is due to an increase in compliance of the sensory-hair attachment to the hair-cell surface.     

Mechanics of the basilar membrane in Caiman crocodilus

Vibration measurements were made at a number of positions near the proximal (basal) end of the basilar membrane, and on the columella footplate, of Caiman crocodilus using a capacitive probe. The measurements established the existence of a mechanical travelling wave in this species. They showed no significant change of mechanical tuning with temperature, and were highly significantly different from previous reports of neural temperature sensitivity (Smolders, J. and Klinke, R. (1984): J. Comp. Physiol. 155, 19-30). Thus the neural sensitivity to temperature change appears not to depend upon basilar membrane mechanics. One interpretation of this is that the basilar membrane passively precedes an active temperature-sensitive filter. It was also found that the limbus supporting the basilar membrane had a measurable, but unturned, vibration and that the effect of draining scala tympani for the measurements was to increase the basilar membrane tuning frequency by a factor of about 1.5.     

Backward Propagation of Otoacoustic Emissions

IntroductionsSounds impinging the eardrum are transmitted viamiddle ear ossicles to the oval window. Stapes vibra-tion creates pressure difference between the scala tym-pani and the scala vestibuli. This pressure differencecauses a movement of cochlear partition and adjacentcochlear fluids. Basilar membrane vibrations result indeflection of hair cell stereocilia, which gate ion chan-nels on their tips. This mechanical-to-electrical trans-duction process converts mechanical vibrations intoelec…     

Low-Frequency Finite-Element Modeling of the Gerbil Middle Ear

The gerbil is a popular species for experimental middle-ear research. The goal of this study is to develop a 3D finite-element model to quantify the mechanics of the gerbil middle ear at low frequencies (up to about 1 kHz). The 3D reconstruction is based on a magnetic resonance imaging dataset with a voxel size of about 45 μm, and an x-ray micro-CT dataset with a voxel size of about 5.5 μm, supplemented by histological images. The eardrum model is based on moiré shape measurements. Each individual structure in the model was assumed to be homogeneous with isotropic, linear, and elastic material properties derived from a priori estimates in the literature. The behavior of the finite-element model in response to a uniform acoustic pressure on the eardrum of 1 Pa is analyzed. Sensitivity tests are done to evaluate the significance of the various parameters in the finite-element model. The Young’s modulus and the thickness of the pars tensa have the most significant effect on the load transfer between the eardrum and the ossicles and, along with the Young’s modulus of the pedicle and stapedial annular ligament, on the displacements of the stapes. Overall, the model demonstrates good agreement with low-frequency experimental data. For example, (1) the maximum footplate displacement is about 35 nm; (2) the umbo/stapes displacement ratio is found to be about 3.5; (3) the motion of the stapes is predominantly piston-like; and (4) the displacement pattern of the eardrum shows two points of maximum displacement, one in the posterior region and one in the anterior region. The effects of removing or stiffening the ligaments are comparable to those observed experimentally.     

Otoacoustic Estimation of Cochlear Tuning: Validation in the Chinchilla

We analyze published auditory-nerve and otoacoustic measurements in chinchilla to test a network of hypothesized relationships between cochlear tuning, cochlear traveling-wave delay, and stimulus-frequency otoacoustic emissions (SFOAEs). We find that the physiological data generally corroborate the network of relationships, including predictions from filter theory and the coherent-reflection model of OAE generation, at locations throughout the cochlea. The results support the use of otoacoustic emissions as noninvasive probes of cochlear tuning. Developing this application, we find that tuning ratios—defined as the ratio of tuning sharpness to SFOAE phase-gradient delay in periods—have a nearly species-invariant form in cat, guinea pig, and chinchilla. Analysis of the tuning ratios identifies a species-dependent parameter that locates a transition between “apical-like” and “basal-like” behavior involving multiple aspects of cochlear physiology. Approximate invariance of the tuning ratio allows determination of cochlear tuning from SFOAE delays. We quantify the procedure and show that otoacoustic estimates of chinchilla cochlear tuning match direct measures obtained from the auditory nerve. By assuming that invariance of the tuning ratio extends to humans, we derive new otoacoustic estimates of human cochlear tuning that remain mutually consistent with independent behavioral measurements obtained using different rationales, methodologies, and analysis procedures. The results confirm that at any given characteristic frequency (CF) human cochlear tuning appears sharper than that in the other animals studied, but varies similarly with CF. We show, however, that the exceptionality of human tuning can be exaggerated by the ways in which species are conventionally compared, which take no account of evident differences between the base and apex of the cochlea. Finally, our estimates of human tuning suggest that the spatial spread of excitation of a pure tone along the human basilar membrane is comparable to that in other common laboratory animals.     

Mechanical tuning of free-standing stereociliary bundles and frequency analysis in the alligator lizard cochlea

Excised cochleae of alligator lizards were prepared to permit microscopic observations of motion of hair-cell free-standing stereociliary bundles and of the underlying basilar papilla during acoustic stimulation, using stroboscopic illumination. In response to tones of frequency from 0.2 to 5 kHz, the papilla rocks about an axis parallel to its length, displacing stereociliary bundles in the morphologically predicted direction of hair cell sensitivity. The papilla moves in phase along its entire length; for frequencies above about 3 kHz, the amplitude of motion of the most basal region is several times larger than that of the rest of the papilla.Over the basal two-thirds of the organ, stereociliary bundles stand freely in endolymph. In this region, maximum bundle height gradually decreases from about 30 to 12 μm; phase vs. frequency characteristics of bundle displacement with respect to the underlying papilla are those of nearly critically damped mechanical resonators. Resonant frequencies measured along the papilla vary inversely with a power (between $\text{3}\text{2}$ and 2) of bundle height and are close in value to auditory nerve fiber CFs measured in vivo at corresponding locations across the nerve. We suggest that length-dependent mechanical tuning of stereociliary bundles determines neural frequency selectivity and tonotopic organization in this part of the organ.     

Similarity of Traveling-Wave Delays in the Hearing Organs of Humans and Other Tetrapods

Transduction of sound in mammalian ears is mediated by basilar-membrane waves exhibiting delays that increase systematically with distance from the cochlear base. Most contemporary accounts of such “traveling-wave” delays in humans have ignored postmortem basilar-membrane measurements in favor of indirect in vivo estimates derived from brainstem-evoked responses, compound action potentials, and otoacoustic emissions. Here, we show that those indirect delay estimates are either flawed or inadequately calibrated. In particular, we argue against assertions based on indirect estimates that basilar-membrane delays are much longer in humans than in experimental animals. We also estimate in vivo basilar-membrane delays in humans by correcting postmortem measurements in humans according to the effects of death on basilar-membrane vibrations in other mammalian species. The estimated in vivo basilar-membrane delays in humans are similar to delays in the hearing organs of other tetrapods, including those in which basilar membranes do not sustain traveling waves or that lack basilar membranes altogether.     

Validity of the Liouville-Green (or WKB) method for cochlear mechanics

This article presents a comparison of Liouville-Green (LG) calculations and exact solutions of 2- and 3-dimensional cochlea models. The agreement is in general quite good. For certain choices of the model parameter's, however, the 2- and 3-dimensional LG solutions show appreciable errors in the region just beyond the location of maximum amplitude of the basilar membrane response. The origin of these errors appears to be the non-uniqueness of the (complex) LG wave number k(x) in 2- and 3-dimensional models: the ‘eikonal equation’ from which k(x) has to be solved has multiple roots. To study this problem somewhat deeper, the properties of the locus of k = k(x) formed when x is varied, are investigated. Erratic behaviour of the LG solution is found to occur when this root locus approaches one of the saddle points of a complex function of k — called Q(k) — which plays the major role in the eikonal equation. Apart from this specific problem, the LG approximation is very well suited to unravel the mechanisms governing wave propagation and attenuation in the cochlea. The analysis shows clearly why and how the response of the basilar membrane builds up to a maximum and which factors cause a turnover and a rapid decrease to occur, in both the long-wave and the short-wave cases. A special discussion is dedicated to the relation between the LG approximation and the absence of wave reflection in cochlea models of the type studied.     

TAK1 Expression in the Cochlea: A Specific Marker for Adult Supporting Cells

Transforming growth factor-β-activated kinase-1 (TAK1) is a mitogen activated protein kinase kinase kinase that is involved in diverse biological roles across species. Functioning downstream of TGF-β and BMP signaling, TAK1 mediates the activation of the c-Jun N-terminal kinase signaling pathway, serves as the target of pro-inflammatory cytokines, such as TNF-α, mediates NF-κβ activation, and plays a role in Wnt/Fz signaling in mesenchymal stem cells. Expression of TAK1 in the cochlea has not been defined. Data mining of previously published murine cochlear gene expression databases indicated that TAK1, along with TAK1 interacting proteins 1 (TAB1), and 2 (TAB2), is expressed in the developing and adult cochlea. The expression of TAK1 in the developing cochlea was confirmed using RT-PCR and immunohistochemistry. Immunolabeling of TAK1 in embryonic, neonatal, and mature cochleas via DAB chromogenic and fluorescent immunohistochemistry indicated that TAK1 is broadly expressed in both the developing otocyst and periotic mesenchyme at E12.5 but becomes more restricted to specific types of supporting cells as the organ of Corti matures. By P1, TAK1 immunolabeling is found in cells of the stria vascularis, hair cells, supporting cells, and Kölliker’s organ. By P16, TAK1 labeling is limited to cochlear supporting cells. In the adult cochlea, TAK1 immunostaining is only present in the cytoplasm of Deiters’ cells, pillar cells, inner phalangeal cells, and inner border cells, with no expression in any other cochlear cell types. While the role of TAK1 in the inner ear is unclear, TAK1 expression may be used as a novel marker for specific sub-populations of supporting cells.     

Ontogenetic changes in the position of hair cell loss after acoustic overstimulation in avian basilar papilla

High intensity sound was used to produce localized hair cell damage within the basilar papilla of chicks at three different ages: embryonic day 20, post-hatch day 10 and post-hatch day 30. At each age separate groups of animals were exposed to broadband white noise or pure tones at 500, 1500 or 3000 Hz for 12 h at 125 dB SPL. Chicks were killed 10 days later. Their basilar papillae were then fixed, dissected free, osmicated, embedded in Epon, sectioned serially and stained. Hair cells were counted at 100 micron intervals throughout the length of the papilla. There was a systematic developmental shift in the position of damage produced by each of the acoustic stimuli. Broadband white noise produced damage only in the basal one half of the cochlea in the embryonic animals while at later ages it produced damage throughout the length of the papilla. Exposure with each of the pure tones produced a discrete area of hair cell loss. However, with each frequency the region of damage shifted apically as a function of the age of the animal at the time of sound exposure. These results suggest that the frequency representation along the basilar papilla is not fixed, but changes during the development of hearing.     

Lateral Wall Histopathology and Endocochlear Potential in the Noise-Damaged Mouse Cochlea

Noise exposure damages the stria and spiral ligament and may contribute to noise-induced threshold shift by altering the endocochlear potential (EP). The aim of this study was to correlate lateral wall histopathology with changes in EP and ABR thresholds. CBA/CaJ mice were exposed to octave band (8–16 kHz) noise for 2 h at intensities ranging from 94 to 116 dB SPL and evaluated 0 h to 8 weeks postexposure. EP in control mice averaged 86 and 101 mV in apical and basal turns, respectively. The 94 dB exposures caused a 40 dB temporary threshold shift (TTS), and there was with no corresponding change in EP. The 112 and 116 dB exposures caused >60 dB threshold shifts at 24 h, and EP was transiently decreased, e.g., to 21 and 27 mV in apical and basal turns after 116 dB. By 1 week postexposure, EP returned to control values in all exposure groups, although those exposed to 112 or 116 dB showed large permanent threshold shifts (PTS). Cochleas were plastic-embedded and serial-sectioned for light microscopic and ultrastructural analysis. Acute changes included degeneration of type II fibrocytes of the spiral ligament and strial edema. The strial swelling peaked at 24 h when significant EP recovery had taken place, suggesting that these changes reflect compensatory volume changes. In the chronic state, massive loss of type II fibrocytes and degeneration of strial intermediate and marginal cells was observed with drastic reduction in membrane surface area. The results suggest that EP shifts do not occur with TTS and also do not add significantly to PTS in the steady state. However, EP loss could contribute to acute threshold shifts that resolve to a PTS. EP recovery despite significant strial degeneration may be partly due to decreased transduction current caused by hair cell damage.