Cochlear Amplifier Research Articles

Synchronized Progression of Prestin Expression and Auditory Brainstem Response during Postnatal Development in Rats.

Prestin is the motor protein expressed in the cochlear outer hair cells (OHCs) of mammalian inner ear. The electromotility of OHCs driven by prestin is responsible for the cochlear amplification which is required for normal hearing in adult animals. Postnatal expression of prestin and activity of OHCs may contribute to the maturation of hearing in rodents. However, the temporal and spatial expression of prestin in cochlea during the development is not well characterized. In the present study, we examined the expression and function of prestin from the OHCs in apical, middle, and basal turns of the cochleae of postnatal rats. Prestin first appeared at postnatal day 6 (P6) for basal turn, P7 in middle turn, and P9 for apical turn of cochlea. The expression level increased progressively over the next few days and by P14 reached the mature level for all three segments. By comparison with the time course of the development of auditory brainstem response for different frequencies, our data reveal that prestin expression synchronized with the hearing development. The present study suggests that the onset time of hearing may require the expression of prestin and is determined by the mature function of OHCs.

Prestin-Dependence of Outer Hair Cell Survival and Partial Rescue of Outer Hair Cell Loss in PrestinV499G/Y501H Knockin Mice.

A knockin (KI) mouse expressing mutated prestin V499G/Y501H (499 prestin) was created to study cochlear amplification. Recordings from isolated outer hair cells (OHC) in this mutant showed vastly reduced electromotility and, as a consequence, reduced hearing sensitivity. Although 499 prestin OHCs were normal in stiffness and longer than OHCs lacking prestin, accelerated OHC death was unexpectedly observed relative to that documented in prestin knockout (KO) mice. These observations imply an additional role of prestin in OHC maintenance besides its known requirement for mammalian cochlear amplification. In order to gain mechanistic insights into prestin-associated OHC loss, we implemented several interventions to improve survival. First, 499 prestin KI’s were backcrossed to Bak KO mice, which lack the mitochondrial pro-apoptotic gene Bak. Because oxidative stress is implicated in OHC death, another group of 499 prestin KI mice was fed the antioxidant diet, Protandim. 499 KI mice were also backcrossed onto the FVB murine strain, which retains excellent high-frequency hearing well into adulthood, to reduce the compounding effect of age-related hearing loss associated with the original 499 prestin KIs. Finally, a compound heterozygous (chet) mouse expressing one copy of 499 prestin and one copy of KO prestin was also created to reduce quantities of 499 prestin protein. Results show reduction in OHC death in chets, and in 499 prestin KIs on the FVB background, but only a slight improvement in OHC survival for mice receiving Protandim. We also report that improved OHC survival in 499 prestin KIs had little effect on hearing phenotype, reaffirming the original contention about the essential role of prestin’s motor function in cochlear amplification.

Corticofugal Modulation of DPOAEs in Gerbils

Efferent auditory feedback on cochlear hair cells is well studied regarding olivocochlear brainstem mechanisms. Less is known about how the descending corticofugal system may shape efferent feedback and modulate cochlear mechanics. Distortion-product otoacoustic emissions (DPOAEs) are a suitable tool to assess outer hair cell function, as they are by-products of the nonlinear cochlear amplification process. The present project investigates the effects of cortical activity on cubic and quadratic DPOAEs in mongolian gerbils, Meriones unguiculatus, through cortical deactivation using the sodium-channel blocker lidocaine. Contralateral cortical microinjections of lidocaine can lead to either an increase or decrease of median DPOAE levels of up to 10.95 dB. The effects are reversible and comparable at all tested frequencies (0.5–40 kHz). They are not restricted to the preferred frequency of the cortical site of injection. Recovery times are between 20 and 120 min depending on stimulation levels and emission type. When the injection is performed in the ipsilateral hemisphere, DPOAE level shifts are lower in amplitude compared to those after injection in the contralateral hemisphere. No significant changes in DPOAE levels are obtained after saline microinjections. Results indicate that deactivation of auditory cortex activity through lidocaine has a considerable impact on peripheral auditory responses in form of DPOAEs, probably through cortico-olivocochlear pathways.

Medial olivocochlear efferent reflex inhibition of human cochlear nerve responses

Inhibition of cochlear amplifier gain by the medial olivocochlear (MOC) efferent system has several putative roles: aiding listening in noise, protection against damage from acoustic overexposure, and slowing age-induced hearing loss. The human MOC reflex has been studied almost exclusively by measuring changes in otoacoustic emissions. However, to help understand how the MOC system influences what we hear, it is important to have measurements of the MOC effect on the total output of the organ of Corti, i.e., on cochlear nerve responses that couple sounds to the brain. In this work we measured the inhibition produced by the MOC reflex on the amplitude of cochlear nerve compound action potentials (CAPs) in response to moderate level (52–60 dB peSPL) clicks from five, young, normal hearing, awake, alert, human adults. MOC activity was elicited by 65 dB SPL, contralateral broadband noise (CAS). Using tympanic membrane electrodes, approximately 10 h of data collection were needed from each subject to yield reliable measurements of the MOC reflex inhibition on CAP amplitudes from one click level. The CAS produced a 16% reduction of CAP amplitude, equivalent to a 1.98 dB effective attenuation (averaged over five subjects). Based on previous reports of efferent effects as functions of level and frequency, it is possible that much larger effective attenuations would be observed at lower sound levels or with clicks of higher frequency content. For a preliminary comparison, we also measured MOC reflex inhibition of DPOAEs evoked from the same ears with f2's near 4 kHz. The resulting effective attenuations on DPOAEs were, on average, less than half the effective attenuations on CAPs.

Consequences of Location-Dependent Organ of Corti Micro-Mechanics.

The cochlea performs frequency analysis and amplification of sounds. The graded stiffness of the basilar membrane along the cochlear length underlies the frequency-location relationship of the mammalian cochlea. The somatic motility of outer hair cell is central for cochlear amplification. Despite two to three orders of magnitude change in the basilar membrane stiffness, the force capacity of the outer hair cell’s somatic motility, is nearly invariant over the cochlear length. It is puzzling how actuators with a constant force capacity can operate under such a wide stiffness range. We hypothesize that the organ of Corti sets the mechanical conditions so that the outer hair cell’s somatic motility effectively interacts with the media of traveling waves—the basilar membrane and the tectorial membrane. To test this hypothesis, a computational model of the gerbil cochlea was developed that incorporates organ of Corti structural mechanics, cochlear fluid dynamics, and hair cell electro-physiology. The model simulations showed that the micro-mechanical responses of the organ of Corti are different along the cochlear length. For example, the top surface of the organ of Corti vibrated more than the bottom surface at the basal (high frequency) location, but the amplitude ratio was reversed at the apical (low frequency) location. Unlike the basilar membrane stiffness varying by a factor of 1700 along the cochlear length, the stiffness of the organ of Corti complex felt by the outer hair cell remained between 1.5 and 0.4 times the outer hair cell stiffness. The Y-shaped structure in the organ of Corti formed by outer hair cell, Deiters cell and its phalange was the primary determinant of the elastic reactance imposed on the outer hair cells. The stiffness and geometry of the Deiters cell and its phalange affected cochlear amplification differently depending on the location.

Cellular and Deafness Mechanisms Underlying Connexin Mutation-Induced Hearing Loss - A Common Hereditary Deafness.

Hearing loss due to mutations in the connexin gene family, which encodes gap junctional proteins, is a common form of hereditary deafness. In particular, connexin 26 (Cx26, GJB2) mutations are responsible for ~50% of non-syndromic hearing loss, which is the highest incidence of genetic disease. In the clinic, Cx26 mutations cause various auditory phenotypes ranging from profound congenital deafness at birth to mild, progressive hearing loss in late childhood. Recent experiments demonstrate that congenital deafness mainly results from cochlear developmental disorders rather than hair cell degeneration and endocochlear potential reduction, while late-onset hearing loss results from reduction of active cochlear amplification, even though cochlear hair cells have no connexin expression. However, there is no apparent, demonstrable relationship between specific changes in connexin (channel) functions and the phenotypes of mutation-induced hearing loss. Moreover, new experiments further demonstrate that the hypothesized K+-recycling disruption is not a principal deafness mechanism for connexin deficiency induced hearing loss. Cx30 (GJB6), Cx29 (GJC3), Cx31 (GJB3), and Cx43 (GJA1) mutations can also cause hearing loss with distinct pathological changes in the cochlea. These new studies provide invaluable information about deafness mechanisms underlying connexin mutation-induced hearing loss and also provide important information for developing new protective and therapeutic strategies for this common deafness. However, the detailed cellular mechanisms underlying these pathological changes remain unclear. Also, little is known about specific mutation-induced pathological changes in vivo and little information is available for humans. Such further studies are urgently required.

Right-ear advantage drives the link between olivocochlear efferent 'antimasking' and speech-in-noise listening benefits.

The mammalian cochlea receives feedback from the brainstem medial olivocochlear (MOC) efferents, whose putative 'antimasking' function is to adjust cochlear amplification and enhance peripheral signal detection in adverse listening environments. Human studies have been inconsistent in demonstrating a clear connection between this corticofugal system and behavioral speech-in-noise (SIN) listening skills. To elucidate the role of brainstem efferent activity in SIN perception, we measured ear-specific contralateral suppression of transient-evoked otoacoustic emissions (OAEs), a proxy measure of MOC activation linked to auditory learning in noisy environments. We show that suppression of cochlear emissions is stronger with a more basal cochlear bias in the right ear compared with the left ear. Moreover, a strong negative correlation was observed between behavioral SIN performance and right-ear OAE suppression magnitudes, such that lower speech reception thresholds in noise were predicted by larger amounts of MOC-related activity. This brain-behavioral relation was not observed for left ear SIN perception. The rightward bias in contralateral MOC suppression of OAEs, coupled with the stronger association between physiological and perceptual measures, is consistent with left-hemisphere cerebral dominance for speech-language processing. We posit that corticofugal feedback from the left cerebral cortex through descending MOC projections sensitizes the right cochlea to signal-in-noise detection, facilitating figure-ground contrast and improving degraded speech analysis. Our findings demonstrate that SIN listening is at least partly driven by subcortical brain mechanisms; primitive stages of cochlear processing and brainstem MOC modulation of (right) inner ear mechanics play a critical role in dictating SIN understanding.

Type II spiral ganglion afferent neurons drive medial olivocochlear reflex suppression of the cochlear amplifier.

The dynamic adjustment of hearing sensitivity and frequency selectivity is mediated by the medial olivocochlear efferent reflex, which suppresses the gain of the ‘cochlear amplifier' in each ear. Such efferent feedback is important for promoting discrimination of sounds in background noise, sound localization and protecting the cochleae from acoustic overstimulation. However, the sensory driver for the olivocochlear reflex is unknown. Here, we resolve this longstanding question using a mouse model null for the gene encoding the type III intermediate filament peripherin (Prph). Prph(−/−) mice lacked type II spiral ganglion neuron innervation of the outer hair cells, whereas innervation of the inner hair cells by type I spiral ganglion neurons was normal. Compared with Prph(+/+) controls, both contralateral and ipsilateral olivocochlear efferent-mediated suppression of the cochlear amplifier were absent in Prph(−/−) mice, demonstrating that outer hair cells and their type II afferents constitute the sensory drive for the olivocochlear efferent reflex.

PKCα-Mediated Signals Regulate the Motile Responses of Cochlear Outer Hair Cells

There is strong evidence that changes in the actin/spectrin-based cortical cytoskeleton of outer hair cells (OHCs) regulate their motile responses as well as cochlear amplification, the process that optimizes the sensitivity and frequency selectivity of the mammalian inner ear. Since a RhoA/protein kinase C (PKC)-mediated pathway is known to inhibit the actin-spectrin interaction in other cell models, we decided to investigate whether this signaling cascade could also participate in the regulation of OHC motility. We used high-speed video microscopy and confocal microscopy to explore the effects of pharmacological activation of PKCα, PKCβI, PKCβII, PKCδ, PKCε, and PKCζ with lysophosphatidic acid (LPA) and their inhibition with bisindolylmaleimide I, as well as inhibition of RhoA and Rho-associated protein kinase (ROCK) with C3 and Y-27632, respectively. Motile responses were induced in isolated guinea pig OHCs by stimulation with an 8 V/cm external alternating electrical field as 50 Hz bursts of square wave pulses (100 ms on/off). We found that LPA increased expression of PKCα and PKCζ only, with PKCα, but not PKCζ, phosphorylating the cytoskeletal protein adducin of both Ser-726 and Thr-445. Interestingly, however, inhibition of PKCα reduced adducin phosphorylation only at Ser-726. We also determined that LPA activation of a PKCα-mediated signaling pathway simultaneously enhanced OHC electromotile amplitude and cell shortening, and facilitated RhoA/ROCK/LIMK1-mediated cofilin phosphorylation. Altogether, our results suggest that PKCα-mediated signals, probably via adducin-mediated inhibition of actin-spectrin binding and cofilin-mediated depolymerization of actin filaments, play an essential role in the homeostatic regulation of OHC motility and cochlear amplification.

Transistor Effect in the Cochlear Amplifier

Abstract The paper presents a new electromechanical amplifying device i.e., an electromechanical biological transistor. This device is located in the outer hair cell (OHC), and constitutes a part of the Cochlear amplifier. The physical principle of operation of this new amplifying device is based on the phenomenon of forward mechanoelectrical transduction that occurs in the OHC's stereocilia. Operation of this device is similar to that of classical electronic Field Effect Transistor (FET). In the considered electromechanical transistor the input signal is a mechanical (acoustic) signal. Whereas the output signal is an electric signal. It has been shown that the proposed electromechanical transistor can play a role of the active electromechanical controlled element that has the ability to amplify the power of input AC signals. The power required to amplify the input signals is extracted from a battery of DC voltage. In the considered electromechanical transistor, that operates in the amplifier circuit, mechanical input signal controls the flow of electric energy in the output circuit, from a battery of DC voltage to the load resistance. Small signal equivalent electrical circuit of the electromechanical transistor is developed. Numerical values of the electrical parameters of the equivalent circuit were evaluated. The range, which covers the levels of input signals (force and velocity) and output signals (voltage, current) was determined. The obtained data are consistent with physiological data. Exemplary numerical values of currents, voltages, forces, vibrational velocities and power gain (for the assumed input power levels below 1 picowatt (10-12 W)), were given. This new electromechanical active device (transistor) can be responsible for power amplification in the cochlear amplifier in the inner ear.

An Electrical Inspection of the Subsurface Cisternae of the Outer Hair Cell

The cylindrical outer hair cell (OHC) of Corti’s organ drives cochlear amplification by a voltage-dependent activation of the molecular motor, prestin (SLC26a5), in the cell’s lateral membrane. The voltage-dependent nature of this process leads to the troublesome observation that the membrane resistor-capacitor filter could limit high-frequency acoustic activation of the motor. Based on cable theory, the unique 30 nm width compartment (the extracisternal space, ECS) formed between the cell’s lateral membrane and adjacent subsurface cisternae (SSC) could further limit the influence of receptor currents on lateral membrane voltage. Here, we use dual perforated/whole-cell and loose patch clamp on isolated OHCs to sequentially record currents resulting from excitation at apical, middle, and basal loose patch sites before and after perforated patch rupture. We find that timing of currents is fast and uniform before whole-cell pipette washout, suggesting little voltage attenuation along the length of the lateral membrane. Prior treatment with salicylate, a disrupter of the SSC, confirms the influence of the SSC on current spread. Finally, a cable model of the OHC, which can match our data, indicates that the SSC poses a minimal barrier to current flow across it, thereby facilitating rapid delivery of voltage excitation to the prestin-embedded lateral membrane.

Tyrosine motifs are required for prestin basolateral membrane targeting.

ABSTRACTPrestin is targeted to the lateral wall of outer hair cells (OHCs) where its electromotility is critical for cochlear amplification. Using MDCK cells as a model system for polarized epithelial sorting, we demonstrate that prestin uses tyrosine residues, in a YXXΦ motif, to target the basolateral surface. Both Y520 and Y667 are important for basolateral targeting of prestin. Mutation of these residues to glutamine or alanine resulted in retention within the Golgi and delayed egress from the Golgi in Y667Q. Basolateral targeting is restored upon mutation to phenylalanine suggesting the importance of a phenol ring in the tyrosine side chain. We also demonstrate that prestin targeting to the basolateral surface is dependent on AP1B (μ1B), and that prestin uses transferrin containing early endosomes in its passage from the Golgi to the basolateral plasma membrane. The presence of AP1B (μ1B) in OHCs, and parallels between prestin targeting to the basolateral surface of OHCs and polarized epithelial cells suggest that outer hair cells resemble polarized epithelia rather than neurons in this important phenotypic measure.

All Three Rows of Outer Hair Cells Are Required for Cochlear Amplification.

In the mammalian auditory system, the three rows of outer hair cells (OHCs) located in the cochlea are thought to increase the displacement amplitude of the organ of Corti. This cochlear amplification is thought to contribute to the high sensitivity, wide dynamic range, and sharp frequency selectivity of the hearing system. Recent studies have shown that traumatic stimuli, such as noise exposure and ototoxic acid, cause functional loss of OHCs in one, two, or all three rows. However, the degree of decrease in cochlear amplification caused by such functional losses remains unclear. In the present study, a finite element model of a cross section of the gerbil cochlea was constructed. Then, to determine effects of the functional losses of OHCs on the cochlear amplification, changes in the displacement amplitude of the basilar membrane (BM) due to the functional losses of OHCs were calculated. Results showed that the displacement amplitude of the BM decreases significantly when a single row of OHCs lost its function, suggesting that all three rows of OHCs are required for cochlear amplification.

Exploiting Dual Otoacoustic Emission Sources.

Two distinct processes generate otoacoustic emissions (OAEs). Reflection-source emissions, here recorded as stimulus frequency OAEs, are optimally informative at low sound levels and are more sensitive to slight hearing loss; they have been linked to cochlear amplifier gain and tuning. Distortion-source emissions are strongest at moderate-high sound levels and persist despite mild hearing loss; they likely originate in the nonlinear process of hair cell transduction. In this preliminary study, we exploit the unique features of each by generating a combined reflection-distortion OAE profile in normal hearing and hearing-impaired ears. Distortion-product (DP) and stimulus-frequency (SF) OAEs were recorded over a broad range of stimulus levels and frequencies. Individual I/O and transfer functions were generated for both emission types in each ear, and OAE peak strength, compression threshold, and rate of compression were calculated. These combined SFOAE and DPOAE features in normal and hearing-impaired ears may provide a potentially informative and novel index of hearing loss. This is an initial step toward utilizing OAE source in characterizing cochlear function and dysfunction.

A model of ionic transport and osmotic volume control in cochlear outer hair cells.

A computational model of the outer hair cell (OHC) of the mammalian cochlea is presented. It addresses the way in which movement of ions controls the cell shape and regulates pH. The model takes into account the possible chloride-bicarbonate exchange function of prestin, a protein highly expressed in the plasma membrane of OHCs. A model of intracellular pH transients following the imposition of a rapid acid load upon the cell has been adapted from the squid axon literature and further extended in order to investigate the effects of ion transport upon the osmotic flux of water into the cell. The model predicts the slow length changes of OHCs reported in the literature a feature which may control the contribution of OHCs to cochlear amplification.

Sound Synthesis with Auditory Distortion Products

Abstract This article describes methods of sound synthesis based on auditory distortion products, often called combination tones. In 1856, Helmholtz was the first to identify sum and difference tones as products of auditory distortion. Today this phenomenon is well studied in the context of otoacoustic emissions, and the “distortion” is understood as a product of what is termed the cochlear amplifier. These tones have had a rich history in the music of improvisers and drone artists. Until now, the use of distortion tones in technological music has largely been rudimentary and dependent on very high amplitudes in order for the distortion products to be heard by audiences. Discussed here are synthesis methods to render these tones more easily audible and lend them the dynamic properties of traditional acoustic sound, thus making auditory distortion a practical domain for sound synthesis. An adaptation of single-sideband synthesis is particularly effective for capturing the dynamic properties of audio inputs in real time. Also presented is an analytic solution for matching up to four harmonics of a target spectrum. Most interestingly, the spatial imagery produced by these techniques is very distinctive, and over loudspeakers the normal assumptions of spatial hearing do not apply. Audio examples are provided that illustrate the discussion.

Prestin is an anion transporter dispensable for mechanical feedback amplification in Drosophila hearing.

In mammals, the membrane-based protein Prestin confers unique electromotile properties to cochlear outer hair cells, which contribute to the cochlear amplifier. Like mammals, the ears of insects, such as those of Drosophila melanogaster, mechanically amplify sound stimuli and have also been reported to express Prestin homologs. To determine whether the D. melanogaster Prestin homolog (dpres) is required for auditory amplification, we generated and analyzed dpres mutant flies. We found that dpres is robustly expressed in the fly’s antennal ear. However, dpres mutant flies show normal auditory nerve responses, and intact non-linear amplification. Thus we conclude that, in D. melanogaster, auditory amplification is independent of Prestin. This finding resonates with prior phylogenetic analyses, which suggest that the derived motor function of mammalian Prestin replaced, or amended, an ancestral transport function. Indeed, we show that dpres encodes a functional anion transporter. Interestingly, the acquired new motor function in the phylogenetic lineage leading to birds and mammals coincides with loss of the mechanotransducer channel NompC (=TRPN1), which has been shown to be required for auditory amplification in flies. The advent of Prestin (or loss of NompC, respectively) may thus mark an evolutionary transition from a transducer-based to a Prestin-based mechanism of auditory amplification.Electronic supplementary materialThe online version of this article (doi:10.1007/s00359-014-0960-9) contains supplementary material, which is available to authorized users.

Connexin26 (GJB2) deficiency reduces active cochlear amplification leading to late-onset hearing loss

Connexin26 (Cx26, GJB2) mutations account for >50% of nonsyndromic hearing loss. The deafness is not always congenital. A large group of these patients (∼30%) demonstrate a late-onset hearing loss, starting in childhood. They have normal hearing early in life and are therefore good candidates for applying protective and therapeutic interventions. However, the underlying deafness mechanism is unclear. In this study, we used a time-controlled, inducible gene knockout technique to knockout Cx26 expression in the cochlea after birth. We found that deletion of Cx26 after postnatal day 5 (P5) in mice could lead to late-onset hearing loss. Similar to clinical observations, the mice demonstrated progressive, mild to moderate hearing loss. The hearing loss initiated at high frequencies and then extended to the middle- and low-frequency range. The cochlea showed normal development and had no apparent hair cell loss. However, distortion product otoacoustic emission (DPOAE) was reduced. The reduction was also progressive and large at high-frequencies. Consistent with DPOAE reduction, we found that outer hair cell electromotility-associated nonlinear capacitance was shifted to the right and the slope of voltage dependence was reduced. The endocochlear potential was reduced in Cx26 conditional knockout (cKO) mice but the reduction was not associated with progressive hearing loss. These data suggest that Cx26 deficiency may impair active cochlear amplification leading to late-onset hearing loss. Our study also helps develop newer protective and therapeutic interventions to this common nonsyndromic hearing loss.

Zebrafish in auditory research: are fish better than mice?

Sensory inner and outer hair cells (IHCs and OHCs) in the mammalian organ of Corti mediate the perception of sound, and hair cells (HCs) in the vestibular system detect head movements. Their characteristic feature is the ‘hair bundle’ of actin-based stereocilia that are arranged in three rows, with increasing length toward the apical cell pole. Sound- or movement-induced deflection towards the longest row opens mechanoelectrical transduction (MET) channels, whereas movement in the opposite direction closes these ion channels. Accordingly, opening and closing of the MET channels transduce sound information and movement of the head into receptor potentials in auditory and vestibular hair cells, respectively. Depolarization induces calcium-dependent neurotransmitter release from specialized presynaptic structures in IHCs and vestibular HCs; the resultant EPSP modulates action potential firing in afferent nerves. In OHCs, receptor potentials produce somatic length changes that comprise the cochlear amplifier and underlie the high sensitivity and frequency selectivity of mammals. The response of the HC membrane potential to MET channel activation is tuned and characterized by a distinct profile of K+ channels in the basolateral membrane (Schwander et al. 2010).

Cytoarchitecture of the mouse organ of corti from base to apex, determined using in situ two-photon imaging.

The cells in the organ of Corti are highly organized, with a precise 3D microstructure hypothesized to be important for cochlear function. Here we provide quantitative data on the mouse organ of Corti cytoarchitecture, as determined using a new technique that combines the imaging capabilities of two-photon microscopy with the autofluorescent cell membranes of the genetically modified mTmG mouse. This combination allowed us to perform in situ imaging on freshly excised tissue, thus minimizing any physical distortions to the tissue that extraction from the cochlea and chemical fixation and staining might have caused. 3D image stacks of the organ of Corti were obtained from base to apex in the cochlear duct, from which 3D lengths and relative angles for inner and outer hair cells, Deiters' cells, phalangeal processes, and inner and outer pillars were measured. In addition, intercellular distances, diameters, and stereocilia shapes were obtained. An important feature of this study is the quantitative reporting of the longitudinal tilts of the outer hair cells towards the base of the cochlea, the tilt of phalangeal processes towards the apex, and Deiters' cells that collectively form a Y-shaped building block that is thought to give rise to the lattice-like organization of the organ of Corti. The variations of this Y-shaped element along the cochlear duct and between the rows of outer hair cells are shown with the third row morphologically different from the other rows, and their potential importance for the cochlear amplifier is discussed.