Cochlear partition anatomy and motion in humans differ from the classic view of mammals.

Mammals detect sound through mechanosensitive cells of the cochlear organ of Corti that rest on the basilar membrane (BM). Motions of the BM and organ of Corti have been studied at the cochlear base in various laboratory animals, and the assumption has been that the cochleas of all mammals work similarly. In the classic view, the BM attaches to a stationary osseous spiral lamina (OSL), the tectorial membrane (TM) attaches to the limbus above the stationary OSL, and the BM is the major moving element, with a peak displacement near its center. Here, we measured the motion and studied the anatomy of the human cochlear partition (CP) at the cochlear base of fresh human cadaveric specimens. Unlike the classic view, we identified a soft-tissue structure between the BM and OSL in humans, which we name the CP "bridge." We measured CP transverse motion in humans and found that the OSL moved like a plate hinged near the modiolus, with motion increasing from the modiolus to the bridge. The bridge moved almost as much as the BM, with the maximum CP motion near the bridge-BM connection. BM motion accounts for 100% of CP volume displacement in the classic view, but accounts for only 27 to 43% in the base of humans. In humans, the TM-limbus attachment is above the moving bridge, not above a fixed structure. These results challenge long-held assumptions about cochlear mechanics in humans. In addition, animal apical anatomy (in SI Appendix) doesn't always fit the classic view.

Molecular organization and fine structure of the human tectorial membrane: is it replenished?

Auditory sensitivity and frequency resolution depend on the physical properties of the basilar membrane in combination with outer hair cell-based amplification in the cochlea. The physiological role of the tectorial membrane (TM) in hair cell transduction has been controversial for decades. New insights into the TM structure and function have been gained from studies of targeted gene disruption. Several missense mutations in genes regulating the human TM structure have been described with phenotypic expressions. Here, we portray the remarkable gradient structure and molecular organization of the human TM. Ultrastructural analysis and confocal immunohistochemistry were performed in freshly fixed human cochleae obtained during surgery. Based on these findings and recent literature, we discuss the role of human TMs in hair cell activation. Moreover, the outcome proposes that the α-tectorin-positive amorphous layer of the human TM is replenished and partly undergoes regeneration during life.

Two-compartment passive frequency domain cochlea model allowing independent fluid coupling to the tectorial and basilar membranes.

The cochlea is a spiral-shaped, liquid-filled organ in the inner ear that converts sound with high frequency selectivity over a wide pressure range to neurological signals that are eventually interpreted by the brain. The cochlear partition, consisting of the organ of Corti supported below by the basilar membrane and attached above to the tectorial membrane, plays a major role in the frequency analysis. In early fluid-structure interaction models of the cochlea, the mechanics of the cochlear partition were approximated by a series of single-degree-of-freedom systems representing the distributed stiffness and mass of the basilar membrane. Recent experiments suggest that the mechanical properties of the tectorial membrane may also be important for the cochlea frequency response and that separate waves may propagate along the basilar and tectorial membranes. Therefore, a two-dimensional two-compartment finite difference model of the cochlea was developed to investigate the independent coupling of the basilar and tectorial membranes to the surrounding liquid. Responses are presented for models using two- or three-degree-of-freedom stiffness, damping, and mass parameters derived from a physiologically based finite element model of the cochlear partition. Effects of changes in membrane and organ of Corti stiffnesses on the individual membrane responses are investigated.

Dual traveling waves in an inner ear model with two degrees of freedom.

We calculate traveling waves in the mammalian cochlea, which transduces acoustic vibrations into neural signals. We use a WKB-based mechanical model with both the tectorial membrane (TM) and basilar membrane (BM) coupled to the fluid to calculate motions along the length of the cochlea. This approach generates two wave numbers that manifest as traveling waves with different modes of motion between the BM and TM. The waves add differently on each mass, producing distinct tuning curves and different characteristic frequencies (CFs) for the TM and the BM. We discuss the effect of TM stiffness and coupling on the waves and tuning curves. We also consider how the differential motions between the masses could influence the cochlear amplifier and how mode conversion could take place in the cochlea.

Inner hair cell stereocilia are embedded in the tectorial membrane.

Mammalian hearing depends on sound-evoked displacements of the stereocilia of inner hair cells (IHCs), which cause the endogenous mechanoelectrical transducer channels to conduct inward currents of cations including Ca2+. Due to their presumed lack of contacts with the overlaying tectorial membrane (TM), the putative stimulation mechanism for these stereocilia is by means of the viscous drag of the surrounding endolymph. However, despite numerous efforts to characterize the TM by electron microscopy and other techniques, the exact IHC stereocilia-TM relationship remains elusive. Here we show that Ca2+-rich filamentous structures, that we call Ca2+ ducts, connect the TM to the IHC stereocilia to enable mechanical stimulation by the TM while also ensuring the stereocilia access to TM Ca2+. Our results call for a reassessment of the stimulation mechanism for the IHC stereocilia and the TM role in hearing.

Tectorial membrane morphological variation: effects upon stimulus frequency otoacoustic emissions.

The tectorial membrane (TM) is widely believed to play an important role in determining the ear's ability to detect and resolve incoming acoustic information. While it is still unclear precisely what that role is, the TM has been hypothesized to help overcome viscous forces and thereby sharpen mechanical tuning of the sensory cells. Lizards present a unique opportunity to further study the role of the TM given the diverse inner-ear morphological differences across species. Furthermore, stimulus-frequency otoacoustic emissions (SFOAEs), sounds emitted by the ear in response to a tone, noninvasively probe the frequency selectivity of the ear. We report estimates of auditory tuning derived from SFOAEs for 12 different species of lizards with widely varying TM morphology. Despite gross anatomical differences across the species examined herein, low-level SFOAEs were readily measurable in all ears tested, even in non-TM species whose basilar papilla contained as few as 50-60 hair cells. Our measurements generally support theoretical predictions: longer delays/sharper tuning features are found in species with a TM relative to those without. However, SFOAEs from at least one non-TM species (Anolis) with long delays suggest there are likely additional micromechanical factors at play that can directly affect tuning. Additionally, in the one species examined with a continuous TM (Aspidoscelis) where cell-to-cell coupling is presumably relatively stronger, delays were intermediate. This observation appears consistent with recent reports that suggest the TM may play a more complex macromechanical role in the mammalian cochlea via longitudinal energy distribution (and thereby affect tuning). Although significant differences exist between reptilian and mammalian auditory biophysics, understanding lizard OAE generation mechanisms yields significant insight into fundamental principles at work in all vertebrate ears.

The supraodontoid space or "apical cave" at the craniocervical junction: a microdissection study.

The extradural supraodontoid space lies anteriorly at the craniocervical junction (CCJ) between the alar ligaments and foramen magnum. It occupies the space between the tectorial and atlanto-occipital membranes. A variety of benign and traumatic lesions may result in neurological compression here with harmful effects. Decompression by the transoral surgical approach often provides relief from these effects. Knowledge of the detailed microanatomy of this space is fragmentary. The purpose of this study was to identify the boundaries and contents of this space by microdissection. Twenty-three en bloc preserved adult cadaveric specimens of the CCJ were dissected to identify the boundaries and contents of the supraodontoid space. The posterior bony elements of the CCJ were removed to enable microdissection (Zeiss DXE Microscope 4-40x) from the tectorial membrane (TM) forwards. The cave-like space faced posteriorly. It had a roof which extended into a wall (anterior atlanto-occipital membrane), a floor (superior surface of the alar ligament), and a mouth covered by the TM. The apical ligament and a thin lining membranous fatty layer divided the cave into a pair of symmetrical halves. The contents, from dorsal to ventral, lay deep to a thin subtectorial membrane. These were the superior fasciculus of the cruciate ligament, a fat-ensheathed knot of plexiform veins (which communicated with the surrounding CCJ vertebral venous plexuses), an arterial arcade between the veins, a pair of fat pads, and branches of the sinuvertebral nerves of the CCJ (lying on the floor). No synovial membrane was found. Knowledge of the anatomy of the apical cave may be of some assistance in transoral (extra- and transdural) surgical approaches to the anterior CCJ region.

Spontaneous Otoacoustic Emissions in TectaY1870C/+ Mice Reflect Changes in Cochlear Amplification and How It Is Controlled by the Tectorial Membrane.

Spontaneous otoacoustic emissions (SOAEs) recorded from the ear canal in the absence of sound reflect cochlear amplification, an outer hair cell (OHC) process required for the extraordinary sensitivity and frequency selectivity of mammalian hearing. Although wild-type mice rarely emit, those with mutations that influence the tectorial membrane (TM) show an incidence of SOAEs similar to that in humans. In this report, we characterized mice with a missense mutation in Tecta, a gene required for the formation of the striated-sheet matrix within the core of the TM. Mice heterozygous for the Y1870C mutation (TectaY1870C/+ ) are prolific emitters, despite a moderate hearing loss. Additionally, Kimura's membrane, into which the OHC stereocilia insert, separates from the main body of the TM, except at apical cochlear locations. Multimodal SOAEs are also observed in TectaY1870C/+ mice where energy is present at frequencies that are integer multiples of a lower-frequency SOAE (the primary). Second-harmonic SOAEs, at twice the frequency of a lower-frequency primary, are the most frequently observed. These secondary SOAEs are found in spatial regions where stimulus-evoked OAEs are small or in the noise floor. Introduction of high-level suppressors just above the primary SOAE frequency reduce or eliminate both primary and second-harmonic SOAEs. In contrast, second-harmonic SOAEs are not affected by suppressors, either above or below the second-harmonic SOAE frequency, even when they are much larger in amplitude. Hence, second-harmonic SOAEs do not appear to be spatially separated from their primaries, a finding that has implications for cochlear mechanics and the consequences of changes to TM structure.

Does the geometrical arrangement of the outer hair cell stereocilia perform a fluid-mechanical function?

CONCLUSIONS: Our experiments qualitatively show that the geometrical structure of the inner ear may have the features of a micro-pump. To further substantiate this hypothesis, additional experiments, particularly on in vivo preparations, are needed. OBJECTIVE: To introduce some new ideas about the functional purpose of the geometric arrangement of the outer hair cell stereocilia. Analogies to some recently developed valveless micro-pumps are pointed out. To illustrate these points, comparative experiments were performed using a simplified macro model. METHODS: Specific structures of the organ of Corti were simulated in a partially open, partially closed acrylic tank. This rough approximation allows the visualization of fluid flows that are generated as a result of the relative motions between the tectorial membrane and the reticular lamina. RESULTS: It was shown that the arrangement of the cochlear elements not only forces fluid to flow in a one-way direction, but also generates a fluid stream that flows through the "outlets" between each two V or W formations of stereocilia. These fluid streams are directed towards the inner hair cells.

Morphology of the basilar papilla of the bobtail lizard Tiliqua rugosa.

The morphology of the basilar papilla of the bobtail lizard was investigated with standard light- and scanning-electron-microscopical methods. The papilla can be subdivided into two parts: a small apical segment which is rather uniform in structure and a long basal segment which displays various systematic changes along its length, for example in the density of the hair cells, the height and shape of the hair-cell stereovillar bundles, the number of stereovilli per bundle and the size of the tectorial structure. In addition, the tectorial structures overlying the two segments are very different in size and morphology. Both tectorial structures are probably sensitive to changes in their ionic environment. The possible functional implications of the papillar morphology described here are discussed with respect to a model of frequency tuning in the bobtail lizard.

Alcian blue staining of cochlear hair cell stereocilia and other cochlear tissues.

Using whole-mount surface preparations of the chinchilla organ of Corti, the anionic characteristics of cochlear tissues was investigated with the polycationic dye Alcian Blue. Our primary goal was to specifically stain the hair cell stereocilia for the purpose of producing cytocochleograms. To specifically stain hair cell stereocilia we investigated the effects of different fixatives and the 'critical electrolyte concentration' [Scott and Dorling (1965) Histochemie 5, 221-233] of MgCl2 on staining specificity. The stereocilia and the tectorial membrane were the most intensely stained cochlear structures and presumably have a strong negative charge. The most important factor in producing specific staining was fixation in osmium tetroxide rather than the addition of an electrolyte to the Alcian Blue staining solution. The use of Alcian Blue produced intense staining of hair cell stereocilia against an unstained background using standard, brightfield light microscopy.

Observations of the sensing and the tectorial membrane in bullfrog amphibian papilla: their possible functional roles.

Three dimensional reconstructions of the amphibian papilla were performed with light microscopic observations, mainly for the sensing membrane (SM). In horizontal sections of the papilla, the anteromedial end of the SM, which makes contact with the massive anterior portion of the tectorial membrane (TM), is several times thicker than the posterolateral end close to the column of the innervating nerves. This gradient of thickness is observed in all the sections from the dorsal portion attached to the TM to the ventral floor of the papilla. The SM connects to the TM in a topological manner; the anteromedial portion of the TM relates to the anterior end of the SM and the anterolateral and the middle portions of the TM correspond to the sites shifting posteriorly on the SM. The morphology of the SM and its manner of connection to the TM suggest that the SM plays important roles in the occurrence of frequency selectivity and of tonotopic organization of the amphibian papilla.

Structural relationships of the unfixed tectorial membrane.

Although the tectorial membrane has a key role in the function of the organ of Corti, its structural relationship within the cochlear partition is still not fully characterised. Being an acellular structure, the tectorial membrane is not readily stained with dyes and is thus difficult to visualise. We present here detailed observations of the unfixed tectorial membrane in an in vitro preparation of the guinea pig cochlea using confocal microscopy. By perfusing the fluid compartments within the cochlear partition with fluorochrome-conjugated dextran solutions, the tectorial membrane stood out against the bright background. The tectorial membrane was seen as a relatively loose structure as indicated by the dextran molecules being able to diffuse within its entire volume. There were, however, regions showing much less staining, demonstrating a heterogeneous organisation of the membrane. Especially Hensen's stripe and regions facing the outer hair cell bundles appeared more condensed. Whereas no connections between Hensen's stripe and the inner hair cell bundles could be observed, there was clearly a contact zone between the stripe and the reticular lamina inside of the inner hair cell.

A quantitative light microscopic study of the bullfrog amphibian papilla tectorium: correlation with the tonotopic organization.

The height, width and cross-sectional area of the bullfrog amphibian papilla tectorial membrane were quantitatively analysed from frontal serial sections. The cross-sectional area (which is a measure of mass) of the tectorium does not appear to be linearly graded along the length of the papilla, but rather spatial gradations occur in more or less a step-wise manner. The spatially graded area (or mass) is well correlated with the width of the tectorium and their relationships with the tonotopic organization of the amphibian papilla are discussed.

Specializations for sharp tuning in the mustached bat: the tectorial membrane and spiral limbus.

The sense of hearing in the mustached bat, Pteronotus parnellii, is specialized for fine frequency analysis in three narrow bands that correspond to approx 30, 60 and 90 kHz constant frequency harmonics in the biosonar signals used for Doppler-shift compensation and acoustic imaging of the environment. Previous studies have identified anatomical specializations in and around the area of the cochlea that processes the dominant second harmonic component, but similar features have not been found in areas related to sharp tuning and high sensitivity for the first or third harmonics. In this report we call attention to the large size of the tectorial membrane and spiral limbus in all three areas that appear to process the harmonically related constant frequency components. These structures are especially pronounced in the regions of the cochlea that respond to the approx 61 kHz, second harmonic and 91.5 kHz, third harmonic bands; they correspond specifically to areas where the density of afferent nerve fibers is high and where very sharply tuned neurons occur. These data for cochleae with multiple specializations lend strong support to the idea that the mass of the tectorial membrane can be an important factor in establishing the response properties of the cochlea.

Auditory stereocilia in the alligator lizard.

The normal anatomy of stereociliary tufts in the basilar papilla of the alligator lizard is described and demonstrated with scanning electron micrographs. Stereociliary tufts in the tectorial region differ from those in the free-standing region in several ways. Tectorial stereociliary tufts are short (less than 10 micron in height), unidirectional in orientation, and covered with a tectorial membrane. Free-standing stereociliary tufts are very tall (up to 38 micron in height), bidirectional in orientation and not covered by a tectorial membrane or any other tectorial substance. The heights of the stereociliary tufts along the length and across the width of the basilar papilla were measured in serial light and transmission electron micrographs. Free-standing stereociliary tufts decrease progressively in height along the length of the basilar membrane, being tallest at the apical end and shortest at the basal end. Tectorial stereociliary tufts do not increase progressively along the length of the basilar membrane but do increase progressively across the width of the basilar membrane, being shortest on the neural side. Free-standing stereociliary tufts are structurally simple being a prominent specialization of the lizard cochlea. Tectorial stereociliary tufts are structurally more complicated conforming more closely to the general anatomical pattern of vertebrate auditory hair cells.

Functional structure of the organ of Corti: a review.

The mammalian auditory organs have a dual sensory system (inner vs. outer hair cells) with distinctly different cellular organizations and innervation patterns. However, the inner (IHCs) and outer (OHCs) hair cells are mechanoreceptors sharing similar general characteristics such as organization of stereocilia (including linkage system) and a gradation of stereociliary height along the length of the cochlea. This gradation of stereociliary height may be the single most important anatomic feature in the tuning capability of the sensory cell. Several lines of evidence suggest that the stereociliary stiffness may be modulated by the sensory cells themselves, most likely via the cuticular plate-rootlet complex. The stereociliary bundles of both types of hair cell are organized in a 'W' formation with a steplike arrangement. In the OHCs, the 'W' formation is sharply angulated and slanted toward the apex, coinciding with the slanted fiber arrangement of the overlying tectorial membrane, which is firmly coupled to the tips of the tallest row of the stereociliary bundles. However, in the IHCs, the 'W' formation is wide and its long axis is linear and arranged at a right angle to the radial axis of the organ of Corti; also, the ciliary bundles are freestanding (with a few exceptions in the basal turn). This arrangement in the IHCs would be best suited for deflection by the radial flow of the endolymph. Present evidence suggests that the subtectorial fluid space exists, is filled with endolymph, and freely communicates with endolymph. Because of the discovery of the phenomenon of 'cochlear emission', the possible motility of the sensory cells, particularly of the OHCs, has drawn intense interest in recent years. Recent investigations with dissociated sensory cells (OHCs) indicate some motile capability under various experimental conditions, although it has not been established that this motility is present in vivo. For this reason, the specialized cellular organization for motility and localization of contractile and cytoskeletal proteins have been investigated. These results support the possibility that the OHCs may have cellular facilities for this function.(ABSTRACT TRUNCATED AT 400 WORDS)

Modifications of cochlear microphonic frequency responses following transient changes of hydrostatic pressure in the perilymph.

Cochlear microphonic potential was recorded with differential electrodes implanted in the various turns of the guinea-pig cochlea. Isointensity frequency responses were plotted in normal conditions and after excessive displacements of the cochlear partition. These displacements were provoked by changes of hydrostatic pressure in the perilymph of scala tympani or scala vestibuli. Typical modifications of the frequency response were observed. The most noticeable was a division in two parts of the response zone which suggested the existence of two resonance peaks. Scanning electron microscopy revealed that changes of hydrostatic pressure provoked alterations of the stereocilia in the outer rows of external hair cells, probably in relation with a decoupling of the tectorial membrane from the organ of Corti. These results are discussed in terms of possible alterations of cochlear micromechanics.

Frequency dependence of synchronization of cochlear nerve fibers in the alligator lizard: evidence for a cochlear origin of timing and non-timing neural pathways.

The dependence of synchronization of spike discharges on tone frequency was measured in cochlear nerve fibers of anesthetized alligator lizards at 21 degrees C. Synchronization measures were based on the fundamental component of a Fourier analysis of the instantaneous discharge rate in response to tone bursts. Measurements were obtained from fibers innervating hair cells in both the region of the cochlea that contains a tectorial membrane (tectorial fibers) and the region where hair-cell stereocilia are free-standing in scala media (free-standing fibers). Both rate and synchronization tuning-curves were measured automatically as a function of tone frequency. For tectorial fibers, the shapes of synchronization tuning-curves are roughly similar to the shapes of rate tuning-curves: the characteristic frequencies (CF's) of both curves are approximately equal. For free-standing fibers, the shape of synchronization tuning-curves differ markedly from those of rate tuning-curves. The CF's of synchronization and rate tuning-curves differ - the ranges are 0.2-0.6 kHz and 1-4 kHz, respectively - and the two CF's are uncorrelated. Synchronization filter-functions, which are contours of synchronization index at constant average discharge rate, were measured as a function of tone frequency for both tectorial and free-standing fibers. These synchronization filter-functions have the shapes of lowpass filters. For the populations of tectorial fibers and of free-standing fibers taken separately, these functions are independent of CF. The corner frequency of these functions is 0.50 +/- 0.038 kHz for tectorial fibers and 0.37 +/- 0.037 kHz for free-standing fibers. We conclude that these populations are characterized by different synchronization filters. For free-standing fibers, synchronization filter-functions measured at average driven discharge rates of about 20 and 40 spikes/s do not differ appreciably, and the high-frequency slope is -80 to -115 dB/decade. The results show that tectorial fibers encode timing information for low-level stimuli, whereas free-standing fibers do not. It is proposed that in the alligator lizard, neural pathways that encode timing information originate in the tectorial region and those that encode non-timing information originate in the free-standing region.

The organ of Corti in the bat Hipposideros bicolor.

The bat Hipposideros bicolor (Hipposideridae, Microchiroptera) is the mammalian species with the highest upper limit of hearing in which the structure of the organ of Corti has been studied. H. bicolor emits pure tone echo-locating signals of 153 kHz, compensates for Doppler shifts in the echo and hears ultrasonic frequencies up to 200 kHz (Neuweiler et al., 1984). The organ of Corti was investigated qualitatively and quantitatively using the technique of semi-thin sectioning. Some complementary ultra-thin sections were also examined. Length, width and cross-sectional area of the basilar membrane, the tectorial membrane, the hair cells with their stereocilia and the organ of Corti were measured at equi-distant positions on the basilar membrane. The organ of Corti of H. bicolor is composed of elements similar to those found in the cochleae of other eutherian mammals studied. However, in H. bicolor some of these elements show species-specific differences when compared to auditorily unspecialized mammals. The most basal region of the cochlea is characterized by miniaturization and re-inforcement of macro- and micro-mechanically important elements. This is interpreted as an adaptation for hearing extremely high frequencies. Specialized structures as well as local maxima of 'normal' elements in the basal and middle cochlear region are associated with evaluation of the echos of emitted pure tones. Besides the basal specializations. Hipposideros also shows specializations in the apical, low frequency, region which can be correlated with passive acoustic orientation.