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PubMed1991

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.

Animals↗
PubMed1998

[Ultrastructure of the tectorial membrane in chick inner ear].

In order to understand the ultrastructure of the tectorial membrane(TM) in avian inner ear and the relations between TM and hair cells and supporting cells, scanning electron microscope(SEM) and transmission electron microscope (TEM) were used to observe the ultrastructure of TM in chick inner ear. The results were that: when viewed under SEM, TM, which possessed a lot of holes inside, was triangular in shape in cross-section and the surface of TM presented fibrous net; when viewed under TEM, TM consisted of fine fibrils and gelatinous substance, only the tallest row of stereocilia and the kinocilium in each hair cell were inserted to TM. The microvilli of supporting cells were anchored to TM by cotton-like fibrous webs. This junction had both elastic and firm, to a certain extent, this kind of junction restricted the movement of TM. According to these structures, there should be different patterns of response to sound stimuli between avian cochlea and mammalian cochlea.

Animals↗
PubMed1990

Tectorial membrane changes in hypothyroid rats during postnatal development.

The morphological changes of the tectorial membrane (TM) during the postnatal development (0, 3, 6, 12 and 25 day old) of the organ of Corti were studied by light microscopy in 20 control and hypothyroid rats. Hypothyroidism was induced by daily administration of propylthioruracil (PTU) until the end of lactation. The auditive receptor in the cochlea of the hypothyroid animals shows serious structural alterations compared with those of normal ones: abnormal persistence of Kölliker's organ, immaturity of sensory cells and supporting cells and a specific distortion of the TM. Differences with controls were first observed on the sixth postnatal day of the hypothyroid rats. The inner spiral sulcus was not shaped and the TM was attached to the Kölliker's organ. In older stages (12 and 25 days), Kölliker's organ was still present. The TM acquired a shap hump with an abnormal fibrillar arrangement in its middle part. It was still attached to the outer supporting cells by a remnant of the marginal net. It was suggested that the TM is secreted by the inner spiral limbus and Kölliker's organ. An abnormal persistence of these structures in the hypothyroidism results in a retardation of Corti's organ development. However, this conclusion does not explain the absence of the outer portion of the TM. Our study confirms the hypothesis that the secretion of any components of the marginal zone of TM is made by outer supporting cells which in PTU-treated animals appear very immature and with hypoplasia.

Animals↗
PubMed1987

Composition and supramolecular organization of the tectorial membrane.

We have shown that collagen accounts for approximately 40% of the total protein of the tectorial membrane (TM) of the guinea pig and have estimated several essential parameters of TM composition including dry weight, wet weight, and water content. The major collagenous protein was definitively identified as type II collagen by SDS-PAGE, CNBr peptide mapping, and immunoblot assays. Quick-freeze, deep-etch electron microscopy of the guinea pig TM demonstrated a dense meshwork of fibers embedded in a complex microfibrillar matrix which may consist of proteoglycans; the larger fibers were similar in size and appearance to type II collagen fibers of elastic cartilage. Finally, the comparative free amino acid profiles of TM strongly suggest that the TM is chemically transparent with respect to endolymph. Thus, the TM appears to consist of a highly hydrated matrix mechanically stabilized by type II collagen fibers.

Amino Acids↗
PubMed1986

Are nonlinearities observed in firing rates of auditory-nerve afferents reflections of a nonlinear coupling between the tectorial membrane and the organ of Corti?

Shear motion between the reticular lamina and the tectorial membrane at the location of the inner hair cells, studied on a network model of the cochlea with a nonlinear coupling between the two structures, exhibits the same type of nonlinearities as seen in the firing rates of auditory-nerve afferents. They include wave form distortions, phase changes and nonmonotonic dependence of the output amplitude on the cochlear input amplitude.

Animals↗
PubMedDate not supplied

Rotated stereociliary bundles and their relationship with the tectorial membrane in the guinea pig cochlea.

Hair cells with rotated stereociliary bundles have been observed in the cochleae of control and kanamycin-treated guinea pigs. The affected outer hair cell bundles have a variable degree of rotation, with some being completely reversed. The inner hair cells are more rarely affected, and only small areas of an individual inner hair cell bundle are abnormal. In counts from ten cochleae, the number of outer hair cells with rotated bundles was most commonly between 10% and 20%, with almost 27% of all outer hair cells affected in the most extreme case. The rotated outer hair cell bundles often have distorted outlines but in other respects closely resemble normal bundles. In particular, they have the usual gradation in stereociliary height, intracellular cross-links and intercellular links to adjacent normally-orientated bundles. There are also corresponding imprints in the tectorial membrane which match the pattern of the stereocilia. In kanamycin-treated guinea pigs, imprints of both normal and rotated hair bundles are present, even when the corresponding bundle is absent, and there are frequently remnants of stereocilia inserted in the imprints. These observations suggest that, apart from their abnormal orientation, the rotated bundles are similar to normal bundles in both their organization and association with the tectorial membrane. The implications of this with respect to transduction and cochlear mechanics are discussed.

Animals↗
PubMed1997

Equilibrium behavior of an isotropic polyelectrolyte gel model of the tectorial membrane: effect of pH.

An isotropic polyelectrolyte gel model (Weiss and Freeman, 1996a) of the tectorial membrane (TM) was extended to incorporate the effect of pH. The effect of pH is analyzed in order to interpret measurements of the effect of pH on the dimensions of the TM (Freeman et al., 1996a). The pH dependence of the model results from the binding of hydrogen ions to TM macromolecules--to both neutral sites with basic pK and negatively charged sites with acidic pK. Parameters of the model can be based on estimates of the concentration in the TM of collagen and glycosaminoglycans (GAGs), two identified constituents of the TM that account for approximately 40% of its dry weight. The resulting model shows swelling responses at both high and low pH that are qualitatively similar to the measurement on the TM but that differ quantitatively. Alternatively, parameters of the model can be chosen in an ad hoc fashion to closely fit the measurements of TM swelling as a function of pH. Taken together these results suggest that either estimates of the collagen and GAG content of the TM are in error or constituents of the TM, which have not yet been identified, contain appreciable pH-dependent fixed charge and contribute to the swelling behavior of the TM.

Binding Sites↗
PubMed1986

Collagen--the predominant protein of the tectorial membrane.

Evidence is presented that, in contrast to the traditional view, a large proportion of the proteins of the tectorial membrane (TM) consists of collagen, primarily type II: characteristic amino acid composition of TM, including high levels of glycine, hydroxyproline and hydroxylysine; comigration of the main TM protein with appropriate collagen standards in two-dimensional gel electrophoresis; digestion of the main TM protein band following treatment with bacterial collagenase; one- and two-dimensional peptide mapping of cyanogen bromide digests of the TM exhibits patterns characteristic for collagen type II.

Amino Acids↗
PubMed2006

Poroelastic bulk properties of the tectorial membrane measured with osmotic stress.

The equilibrium stress-strain relation and the pore radius of the isolated tectorial membrane (TM) of the mouse were determined. Polyethylene glycol (PEG), with molecular mass (MM) in the range 20-511 kDa, added to the TM bathing solution was used to exert an osmotic pressure. Strain on the TM induced by isosmotic PEG solutions of different molecular masses was approximately the same for MM > or = 200 kDa. However, for MM < or = 100 kDa, the TM strain was appreciably smaller. We infer that for the smaller molecular mass, PEG entered the TM and exerted a smaller effective osmotic pressure. The pore radius of the TM was estimated as 22 nm. The equilibrium stress-strain relation of the TM was measured using PEG with a molecular mass of 511 kDa. This relation was nonlinear and was fit with a power function. In the radial cochlear direction, the transverse stiffness of the TM was 20% stiffer in the inner than in the outer region. TM segments from the basal region had a larger transverse stiffness on average compared to sections from the apical-middle region. These measurements provide a quantitative basis for a poroelastic model of the TM.

Animals↗
PubMed1979

The effect of changes in endolymphatic ion concentrations on the tectorial membrane.

The development of a new preparation technique has allowed the effects of fluid-substitution experiments on the tectorial membrane (t.m.) to be studied morphologically and not only, as previously, electrophysiologically. The organ of Corti and t.m. were examined in situ, unfixed and with in vivo-like ionic conditions under light-microscopical control (differential-interference-contrast, magnification 400--800X). An irreversible shrinkage of the marginal and middle zones of the t.m. was observed under the influence of Na+ ions (substitution of endolymph with artificial perilymph). This shrinkage is also seen in specimens prepared for scanning electron microscopy after perfusion of scale media with perilymph. If endolymph is replaced by an isotonic solution of EDTA (ethylenediaminetetraacetate) the t.m. can swell in its vestibulo-tympanal extent, e.g. from 39 to 72 microns. This swelling is reversible on addition of CaCl2. Although the size of the t.m. is not influenced to any great extent by changes in fluid osmolarity, reduction of H+ ion concentration leads to a minimal state of hydration of the protobril system at pH 4.3 (i.e. region of isoelectric point). The mechanism of Na+ and Ca2+ effects are discussed. These results indicate that the subtectorial space is morphologically separated from the scala media proper by the marginal zone.

Animals↗
PubMed1998

Type A fibril of the mouse tectorial membrane shows D-periodicity: an atomic force microscopic examination.

This study demonstrated that type A fibrils of the mouse tectorial membrane showed a morphology characteristic of collagen, as demonstrated using an atomic force microscope. In the topographical imaging mode, the surface of the type A fibrils showed a periodic pattern, consisting of alternating grooves and ridges. The periodicity of the type A fibrils was 69.1 +/- 0.6 nm, which is in accordance with characteristic collagen D-periodicity. The difference in height between grooves and ridges was 1.6 +/- 0.3 nm. In the variable deflection imaging mode, the type A fibrils showed a clear banding pattern, which consisted of alternating light and dark zones, with D-periodicity. In addition, the type A fibrils exhibited one minor dark band in the light zone and one minor light band in the dark zone.

Aged↗
PubMed1990

Localization of anionic sulfate groups in the tectorial membrane.

Colloidal iron hydroxide (CIH) staining demonstrates the existence of anionic sulfate groups of glycoconjugates associated with several constituents of the tectorial membrane (TM). In the adult animal, labelling in the main body of the TM appears as long, electron-dense patches surrounding type A fibrils which show alternating stained and unstained zones. On the other hand, labelling of the fibrils of the matrix of the TM appears as single, CIH particles with no special arrangement. Some of the structurally distinct regions of the TM are also labelled (limbal zone, Hensen's stripe and inner portions of the cover net), while others are not (marginal band and outer portions of the cover net). Staining of type A fibrils in the major TM is already present in newborn animals; while, both the outermost region of the TM closest to the cells of the organ of Kölliker and the minor TM are not labelled. The implications of these distributions of sulfated glycoconjugates for the electrochemical properties of the TM are discussed.

Animals↗
PubMed1995

The osmotic response of the isolated, unfixed mouse tectorial membrane to isosmotic solutions: effect of Na+, K+, and Ca2+ concentration.

Changes in the size, shape, and structure of the isolated tectorial membrane (TM) of the mouse were measured in response to isosmotic changes in the ionic composition of the bathing solution. Substitution of artificial perilymph (AP) for artificial endolymph (AE) caused a small (approximately 1%) shrinkage of the TM's thickness. This substitution alters not only the predominate cation (from K+ to Na+) but also the Ca2+ concentration (from 20 mumol/l to 2 mmol/l). When the predominate cation was changed from K+ to Na+, while holding Ca2+ concentration constant, results depended on Ca2+ concentration: there was a small (approximately 1%) swelling for 20 mumol/l Ca2+, larger (approximately 14%) swelling for lower (< 7 mumol/l) concentrations of Ca2+, and little response for 2 mmol/l Ca2+ or for solutions containing the Ca2+ chelator EGTA. Addition of Ca2+ while holding the predominate cation constant caused shrinkage of the TM; both removal of Ca2+ and addition of the Ca2+ chelator EGTA caused swelling. Swelling responses were largely reversible if the magnitude of the swelling was small. Responses greater than a few percent were only partially reversible and caused long-lasting changes. Changes in ionic composition of the bath affected not only the thickness of the TM but also its other dimensions. Solution changes that increase TM thickness tend to cause radial shearing motions of the surfaces of the TM, which are accompanied by small decreases in width. Little change in length was observed. Although the responses were non-isotropic, increases in thickness were highly correlated with increases in volume. Swelling of the TM was also accompanied by a reduction in prominence of its radially oriented fibrillar structure. These results for the isolated TM of the mouse are qualitatively similar to those obtained previously for the isolated chick TM (Freeman et al., 1994) but different from those obtained for the in vitro mouse TM (Kronester-Frei, 1979a).

Acoustic Stimulation↗
PubMed1995

Secretion of a new basal layer of tectorial membrane following gentamicin-induced hair cell loss.

Scanning electron microscopy (SEM) and video-enhanced DIC light microscopy were used to assess morphological changes in the chick tectorial membrane (TM) following gentamicin-induced hair cell loss. Gentamicin was administered (100 mg/kg/day for 3 days) and isolated and in-situ TMs were examined in both fixed and unfixed preparations at days 5 and 10 after the initial injection. Although this protocol induced hair cell damage extending up to 75% of the length of the basilar papilla, there was no apparent damage to the TM itself. However, the ejection of damaged hair cells appeared to sever the filamentous attachments between the TM and the apical surface of the basilar papilla. In SEM preparations this detachment caused the TM to shrink back toward the superior edge. Interestingly, despite the lack of TM damage, gentamicin treatment did reveal the secretion of a new basal layer of TM. Secretion of this new basal layer had begun by day 5 and it was well organized by day 10. This new layer formed attachments to both the recovering basilar papilla and the overlying original TM, a step thought to be necessary for the restoration of auditory function in the regenerating cochlea.

Animals↗
PubMed1995

Tectorial membrane regeneration in acoustically damaged birds: an immunocytochemical technique.

A novel immunocytochemical method was used to determine whether the sound-damaged adult quail ear can repair its tectorial membrane (TM) and to compare the repair in quail to that in chicks. Birds were exposed to an octave band noise with a center frequency of 1.5 kHz at 116 dB SPL for 4 h. The chicks were grouped based on recovery duration (0 and 7 days), while the quail were divided into 0-, 7-, and 14-day recovered groups. At the end of the recovery period, the animals were sacrificed, and their basilar papillae labeled with a TM-specific monoclonal primary antibody solution followed by a diaminobenzidine process. Examinations under a stereoscope revealed that a patch lesion devoid of TM was located on all 0-day recovered papillae. Seven days later, a honeycomb-patterned layer was observed covering the lesion. In 14-day recovered quail ears, the honeycomb layer appeared similar to that seen at 7 days post-exposure. These observations indicated that both chicks and quail were able to repair their TM within 7 days following exposure to intense sound.

Acoustic Stimulation↗
PubMed1996

Further studies on the mechanics of the cochlear partition in the mustached bat. I. Ultrastructural observations on the tectorial membrane and its attachments.

From semithin and ultrathin sections of the mustached bat cochlea, baso-apical gradients in ultrastructural composition, shape and attachment site of the tectorial membrane (TM) were determined in relation to gradients in hair cell size and stereocilia size. These provide a data base for estimates of the mechanical properties of the organ of Corti as they relate to specialized aspects of the cochlear frequency map (Kössl and Vater, 1996). As in other mammals, the TM is composed to type A and type B protofibrils. Measurements of the packing density of type A protofibrils reveal gradients in both the radial and longitudinal direction. Distinct variations in packing density of type A protofibrils across the radial extent of the TM allow the definition of more subregions than in other mammals. Throughout the cochlea, packing density is highest in the 'stripe' region located close to the spiral limbus. The centrally located 'core' region of the middle zone contains distinctly fewer type A protofibrils than the laterally located 'mantle' region of the middle zone. The TM in the specialized basal turn (first and second half-turns) features a higher packing density of type A protofibrils in the 'mantle' than the TM in the apical cochlea (upper third to fifth half-turns), and in incorporation of longitudinally directed type A protofibrils in the marginal zone. Among cochlear turns, there are pronounced changes in cross-sectional area of the TM and the extent of its limbal attachment site. Within the densely innervated second half-turn that contains an expanded representation of the 60 kHz constant frequency (CF) component of the echolocation signal, both the cross-sectional area (see also Henson and Henson, 1991) and the attachment site of the TM are enlarged. An extended limbal attachment site is also observed in the densely innervated region of the lower first half-turn that represents the upper harmonics of the call. Within the sparsely innervated region of the upper first half-turn, the limbal attachment site of the TM is significantly diminished. Size of outer hair cells (OHC) ranges between 12 and 13 microns throughout the basal 80% of cochlear length and reaches maximal values of 20 microns in the apex. Size of OHC stereocilia ranges between 0.7 and 0.8 microns throughout the basal 60% of cochlear length and reaches a maximal size of 2.2 microns in the apex. These data corroborate and extend previous notions that morphological specializations of the TM in concert with specializations of the basilar membrane and perilymphatic spaces play an integral role in creating specialized cochlear tuning in the mustached bat.

Animals↗
PubMed2003

Role of the tectorial membrane revealed by otoacoustic emissions recorded from wild-type and transgenic Tecta(deltaENT/deltaENT) mice.

Distortion product otoacoustic emissions (DPOAE) were recorded from wild-type mice and mutant Tecta(deltaENT/deltaENT) mice with detached tectorial membranes (TM) under combined ketamine/xylaxine anesthesia. In Tecta(deltaENT/deltaENT) mice, DPOAEs could be detected above the noise floor only when the levels of the primary tones exceeded 65 dB SPL. DPOAE amplitude decreased with increasing frequency of the primaries in Tecta(deltaENT/deltaENT) mice. This was attributed to hair cell excitation via viscous coupling to the surrounding fluid and not by interaction with the TM as in the wild-type mice. Local minima and corresponding phase transitions in the DPOAE growth functions occurred at higher DPOAE levels in wild-type than in Tecta(deltaENT/deltaENT) mice. In less-sensitive Tecta(deltaENT/deltaENT) mice, the position of the local minima varied nonsystematically with frequency or no minima were observed. A bell-like dependence of the DPOAE amplitude on the ratio of the primaries was recorded in both wild-type and Tecta(deltaENT/deltaENT) mice. However, the pattern of this dependence was different in the wild-type and Tecta(deltaENT/deltaENT) mice, an indication that the bell-like shape of the DPOAE was produced by a combination of different mechanisms. A nonlinear low-frequency resonance, revealed by nonmonotonicity of the phase behavior, was seen in the wild-type but not in Tecta(deltaENT/deltaENT) mice.

Acoustic Stimulation↗