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PubMed2003

The inhibition of cyclin-dependent kinases induces differentiation of supernumerary hair cells and Deiters' cells in the developing organ of Corti.

In the embryonic day 19 organs of Corti, we showed that roscovitine, a chemical inhibitor of cyclin-dependent kinases (CDKs), significantly increased the number of hair cells (HCs) and corresponding supporting cells (SCs) by triggering differentiation of precursor cells without interacting with cell proliferation. The effect of roscovitine was mimicked by other CDK1, 2, 5, and 7 inhibitors but not by CDK4/6 and mitogen-activated protein kinase pathway antagonists. Immunohistochemical analysis indicated that roscovitine-specific intracellular targets, CDK1, 2, 5, and 7, were expressed in the organ of Corti and especially in Hensen's cells. Affinity chromatography studies showed a tight correlation between the protein levels of CDK1/2 and 5 and the rate of roscovitine-induced supernumerary cells in the organ of Corti. In addition, we demonstrated that basal CDK activity was higher and more roscovitine-sensitive at developmental stages that are selectively permissive for the emergence of supernumerary cells. These results suggest that CDKs are involved in the normal development of the organ of Corti and that, at least in E19 embryos, inhibition of CDKs is sufficient to trigger the differentiation of HCs and corresponding SCs, presumably from the Hensen's cell progenitors and/or from progenitors located in the greater epithelial ridge area.

Animals↗
PubMed1999

Expression of membrane-bound and cytosolic guanylyl cyclases in the rat inner ear.

Membrane-bound guanylyl cyclases (GCs) are peptide hormone receptors whereas the cytosolic isoforms are receptors for nitric oxide. In the inner ear, the membrane-bound GCs may be involved in the regulation of fluid homeostasis and the cytosolic forms possibly play a role in signal processing and regulation of local blood flow. In this comprehensive study, we examined, qualitatively and quantitatively, the transcription pattern of all known GC isoforms in the inner ear from rat by RT-PCR. The tissues used were endolymphatic sac, stria vascularis, organ of Corti, organ of Corti outer hair cells, cochlear nerve, Reissner's membrane, vestibular dark cells, and vestibular sensory cells. We show that multiple particulate (GC-A, GC-B, GC-D, GC-E, GC-F and GC-G) and several subunits of the heterodimeric cytosolic GCs (alpha1, alpha2, beta1 and beta2) are expressed, albeit at highly different levels. GC-C was not found. GC-A and the soluble subunits alpha1 and beta1 were transcribed ubiquitously. GC-B was present in all tissues except stria vascularis, which contained GC-A and traces of GC-E and GC-G. GC-B was by far the predominant membrane-bound isoform in the organ of Corti (86%), Reissner's membrane (75%) and the vestibulum (80%). Surprisingly, GC-E, a retinal isoform, was detected in significant amounts in the cochlear nerve (8%) and in the organ of Corti (4%). Although the cytosolic GC is a heterodimer composed of an alpha and a beta subunit, the mRNA transcription of these subunits was not stoichiometric. Particularly in the vestibulum, the transcription of the beta1 subunits was at least four-fold higher than of the alpha1 subunit. The data are compatible with earlier suggestions that membrane receptor GCs may be involved in the control of inner ear electrolyte and fluid composition whereas NO-stimulated GC isoforms mainly participate in the regulation of blood flow and supporting cell physiology.

Animals↗
PubMed1999

Retinoic acid signaling is necessary for the development of the organ of Corti.

The cellular mosaic of the mammalian organ of Corti represents one of the most highly ordered structures in any vertebrate system. A single row of inner hair cells and three or four rows of outer hair cells extend along the basal-to-apical axis of the cochlea. The factors that play a role in the development of specific cell types within the cochlea are largely unknown; however, the results of previous studies have strongly suggested that retinoic acid plays a role in the development of cells as hair cells. To determine whether cochlear progenitor cells can respond directly to retinoic acid, the expression patterns for each of the RAR and RXR receptors within the embryonic cochlear duct were determined by in situ hybridization. Results indicate that RARalpha, RXRalpha, and RXRgamma are initially expressed throughout the cochlear duct. As development continues, the expression of each receptor becomes more intense in cells that will develop as hair cells. At the same time, receptor expression is down-regulated in cells that will develop as nonsensory cell types. To determine the effects of retinoic acid signaling during the development of the organ of Corti, activation of retinoid receptors was blocked in cultures of the embryonic cochlea through receptor-specific antagonism or inhibition of retinoic acid synthesis. Results indicate that inhibition of retinoic acid signaling induces a significant decrease in the number of cells that develop as hair cells and a disruption in the development of the organ of Corti. These results demonstrate that cells within the developing cochlea can respond to retinoic acid and that signaling by retinoic acid is necessary for the normal development of the organ of Corti.

Animals↗
PubMed1995

Cytological changes related to maturation of the organ of Corti and opening of Corti's tunnel.

Maturation of the organ of Corti in the gerbil was analyzed between 2 and 16 days after birth (DAB) by electron microscopy and immunostaining for beta-tubulin. At 2 DAB, the organ of Corti consisted of stratified epithelium bearing immature sensory hair cells (HCs) and supporting cells. Maturation of OHCs and Deiters cells progressed in a medial-to-lateral direction and cytoskeletal development in inner pillar cells preceded that in outer pillar cells at the single location studied along the frequency-place map. Pillar cell differentiation progressed through a unique stage characterized by the appearance and stratification of structural features apparently concerned with opening of Corti's tunnel and subsequently showed other structural changes related to maturity toward the adult form. Development of the microtubule cytoskeleton occurred first in the cell's apex and proceeded basally. Ruffling of a middle region of the cell surface by microvilli appeared to promote separation between inner and outer pillar cells and initiate tunnel opening at 4 DAB. Proliferation of distended cisternae of granular reticulum evidenced proteinaceous secretion by these cells between 4 and 8 DAB. Subsequent tunnel expansion at about 14 DAB coincided with appearance in outer pillar cells of tubulocisternal endoplasmic reticulum and associated Golgi complexes that are thought to mediate fluid and ion secretion. Sixteen days postnatally after disappearance of granular and tubulocisternal reticula and Golgi complexes and at the time of clearing of tunnel fluid, lysosomes interpreted as mediating catabolism of endocytosed protein congregated beneath the apical and apicolateral plasmalemmae of inner pillar cells. As with pillar cells, development of the microtubule system in Deiters cells proceeded from the cell's apex to base. Following differentiation of their microtubule system by 8 DAB, Deiters cells showed expansion of Golgi cisternae between 10 and 15 DAB and development of tubulocisternal endoplasmic reticulum at 15 DAB. Hair cells possessed abundant, distinctively large mitochondria from 4 to 10 DAB. The subsurface cisternae matured earlier in medial as opposed to lateral outer hair cells. Vesicles budding from underlying cisternae appeared associated with development of subsurface cisternae and at 16 DAB were still observed in third row but not in more mature first row HCs.

Animals↗
PubMed1976

[Thermal noise of Corti's organ hair cells].

Thermal agitation of the hair cells stereocilia of Corti organ is estimated. It is shown that with threshold sound stimulus the signal-to-noise ratio for single stereocilium and for single hair cell is less than 1. Some methods are proposed for signal-noise discrimination.

Auditory Perception↗
PubMed1979

Noise-noise effect upon the spreading of the posttraumatic progressive necrosis in the organ of Corti.

Progressive necrosis of the organ of Corti caused by a disturbance of the ionic composition of the Cortilymph resulting from the infiltration of endolymph into the organi of Corti after traumatization of its barrier against the scala media, is greatly intensified by posttraumatic stimulation with noise of physiological intensities. This effect is attributed to oscillation of the cochlear partition, with resultant greater blending of the Cortilymph and the endolymph. Stimulation augments necrosis fourfold towards the base of the cochlea and twofold towards the helicotrema. Posttraumatic stimulation is one of the factors which markedly potentiate the regressive effect of acoustic trauma.

Acoustic Stimulation↗
PubMed1981

Scanning electron microscopy of the human cochlea--the organ of Corti.

The surface of the organ of Corti from normally hearing adult humans has been examined with the scanning electron microscope. It is possible to construct cytocochleograms and to derive a regression line with confidence limits to represent the distribution of the sensory hair cells. Examining individual hair cells more closely, the number of cilia on each hair cell, decreased linearly with distance, from the base of the cochlea. However, the length of the longest cilia on each outer hair cell increased linearly with distance.

Adult↗
PubMed1994

Stimulus biasing: a comparison between cochlear hair cell and organ of Corti response patterns.

Responses from the organ of Corti (OC) fluid space and from individual hair cells are collected for short duration tone pips measured alone and in the presence of a 20 Hz bias tone. Because of the relatively long period of the acoustic bias signal, a probe can be selectively placed at precise locations within a single response period. Since bias effects produced during maximum basilar membrane velocity are negligible, this report documents changes associated with basilar membrane displacements to scala vestibuli and scala tympani. Hair cell response patterns are compared with those measured nearby in the OC to determine the degree to which inner (IHC) and outer hair cells (OHC) contribute to the gross cochlear potentials. It is shown that intracellular OHC and OC dc responses are strongly influenced by the bias while intracellular IHC dc responses are minimally affected. Although an association has been established between the cochlear microphonic and outer hair cell ac receptor potentials, the well correlated changes in the outer hair cell dc receptor potential and the organ of Corti summating potential suggest that gross cochlear potentials reflect dc as well as ac contributions from nearby OHCs. Since IHC responses are not influenced by the bias, they appear to contribute less than OHCs to extracellular potentials. These conclusions, however, are restricted to the moderately high input levels required to produce these effects (Durrant and Dallos, 1974) and to the central region of the cochlea where the recordings are made. Finally, the degree to which bias effects are contaminated by two-tone suppression is discussed.

Acoustic Stimulation↗
PubMed1997

Postnatal maturation of the organ of Corti in gerbils: morphology and physiological responses.

The organ of Corti, the sensory epithelium of hearing in mammals, matures postnatally in the gerbil. Quantitative analyses of the postnatal development of the organ of Corti, including supporting cells and the basilar membrane, were carried out. The morphological study confirmed that maturation of the sensory cells proceeds with a base-to-apex gradient, with the outer hair cells appearing to mature before the inner hair cells. Maturation of the supporting cells and the basilar membrane commenced first in the middle turn. Expansion of the second row of Deiters' cells began at 6 days after birth in the middle turn, before enlargement of the pillar cell heads at 8 days postnatally. Pillar cell head enlargement continued until 20 days postnatally in the middle turn. The tunnel of Corti and spaces of Nuel appeared first in the middle turn between 8 and 10 days postnatally. The maturation of the basilar membrane involved the thickening of the central hyaline layer and a reduction in the epithelial cells on the tympanic aspect. This process continued until about 20 days after birth. The cochlear microphonic potential, whole nerve action potential, and stimulus frequency otoacoustic emissions were recorded from 12 days after birth onward and related to changes in organ of Corti morphology. The results show that changes in the accessory structures continue throughout the period of onset and development of cochlear responses between 12 and 20 days after birth, and may therefore influence the micromechanical responses of the organ of Corti to acoustic stimuli during this period.

Animals↗
PubMedDate not supplied

Observations of the organ of Corti under in vivo-like conditions.

The organ of Corti of the rat has been investigated within minutes after death in different fluids bathing the organ of Corti. Static compliance of the tectorial membrane across the membrane was measured by microsprings, and the response of the membrane to a vibrating micro-electrode was studied. In addition, fluid motion about the organ caused by the placement of a vibrating micro-electrode on the tectorial membrane was observed. The static compliance of the tectorial membrane was unaffected by the bathing solutions. Dynamic characteristics of the membrane could be completed altered by the addition of aldehyde solutions. As the cochleas used by von Békésy (1960) were preserved in formaldehyde solutions, it is concluded that his observations regarding the dynamic characteristics of the tectorial membrane were in part artifact. It is suggested that a proper description of the mechanisms of the organ of Corti must include a tectorial membrane whose limbal attachment allows for both rotation and displacement. Such a description is presented. A streaming of fluid was observed directed from Hensen's cells toward the cochlea wall. Possible mechanisms for the generation of this streaming are discussed.

Animals↗
PubMed2001

Direct evidence of nitric oxide production in the guinea pig organ of Corti.

Production of nitric oxide (NO) in the organ of Corti of the guinea pig was investigated using the new fluorescence indicator 4,5-diaminofluorescein diacetate for direct detection of NO. The organ of Corti, lateral wall of the cochlea and isolated outer and inner hair cells were examined to locate NO production sites. The fluorescence intensities were augmented by stimulation with L-arginine or glutamate, and significantly increased after inoculation with lipopolysaccharide. This is the first direct evidence of NO production in the cochlea. NO may play an important role in the physiology of the organ of Corti and may also be involved in hearing disorders.

Animals↗
PubMed1981

The combination of scanning and transmission electron microscopy techniques in pathology of the organ of Corti in guinea pigs.

The pathology of the organ of Corti in guinea pigs is studied by means of SEM and TEM. It was the purpose of this study to examine the surface of an experimentally altered organ of Corti by means of SEM screening method). As it was our main interest to find out the localization of acoustic stimulation regarding the tonotopicity on the surface of the organ of Corti, we always examined the whole cochlear by means of SEM first. In acoustic experiments we used long-time pure-tone stimulation beyond the directly damaging sound pressure level, as well as white noise or shots for experimental damage of hair cells and supporting cells. Specimens of particular interest underwent a special procedure for a supplementary TEM examination. A prolonged and modified technique of embedding the specimens in Epon 812 permitted us to obtain supplementary data from TEM without major artifacts in the fine structure of the cells.

Acoustic Stimulation↗
PubMed2000

Characterization of novel and identified genes in guinea pig organ of corti.

A number of proteins are expressed in the organ of Corti and are considered to be responsible for hearing. However, most of them have not been identified. Therefore, to achieve a better understanding of the genetic factors influencing these traits, the first step is to characterize the genes expressed in the organ of Corti. In the present study, a cDNA library was constructed from the guinea pig organ of Corti. After sequencing isolated clones, 196 expressed sequence tags (ESTs) were identified with FASTA analysis: 65 ESTs showed significant sequence homology to previously identified genes in guinea pig, human or other species, and 131 ESTs showed no significant matches to sequences already present in the DNA database DDBJ/GenBank/EMBL. A variety of matching sequences, some of which were known to be cochlea-specific, were found through FASTA analysis of the 65 clones. RT-PCR with a panel of 10 different tissue mRNA revealed the restricted expression of 13 unknown clones. The results of our analysis allowed the establishment of a list of genes expressed in the guinea pig organ of Corti.

Animals↗
PubMed2003

Organ of Corti kinematics.

The internal workings of the organ of Corti and their relation to basilar membrane motion are examined with the aid of a simple kinematic model. It is shown that, due to the lever system embodied in the organ of Corti, there is a significant transformer gain between basilar membrane and cilia displacements. While this transformation is nonlinear, linear response prevails in the narrow physiologically relevant operating range of the ciliary transducer. The model also simulates cilia deflection when the mechanical stimulus is the length change of outer hair cells.

Animals↗
PubMed1977

Freeze-fracture studies on the synapses in the organ of Corti.

Receptoneural junctions and synapses in the organ of Corti of the chinchilla have been examined with the freeze-fracture technique. The presynaptic membranes at the receptoneural junctions of inner and outer hair cells have many structural features in common with membranes found at chemical synapses outside the organ of Corti. However, the membranes of the postsynaptic afferent terminals are quite different depending on whether they are part of an inner or outer hair cell synapse. These differences in the distribution of intramembrane particles suggest that the transmitters, or transmitter actions, may be different at these two synapses. The distribution of particles in the postsynaptic membrane at efferent synapses with outer hair cell differs from that in the postsynaptic membrane at efferent synapses with afferent terminals or fibers, suggesting that transmitter actions at these locations could also differ.

Animals↗
PubMed1997

Structure of glycogen particles in organ of Corti's outer hair cells in three rodent species.

Organ of Corti's outer hair cells are one of the few cell types in mammals to contain large cytoplasmic glycogen stores, and the only one in the adult auditory receptor. Previous reports on the structure and distribution of glycogen in the adult organ of Corti were mainly based on light microscopy histo- and cytochemical methods, and the scare EM studies on the topic relied on techniques which were not sensitive or specific enough. Furthermore, it has been reported that glycogen particles are not present in outer hair cells of all species. A first goal of the present study was to describe the ultrastructure of glycogen stores in organ of Corti's outer hair cells in Guinea pig, rat, and mouse, using the periodic acid-thiocarbohydrazide-silver proteinate method. In addition, differences in the subcellular and cochleotopic distribution of this substance were analyzed. In the adult organ of Corti only the outer hair cells contain glycogen stores. Present throughout their cytoplasm, these deposits appear either as single beta particles, or as aggregates of these, forming alpha particles. Though most alpha particles are round, some appear long and conspicuously straight in longitudinal sections of those cells near the apex of the cochlea, and they seem to be apposed to some filamentous structure. On the other hand, when the cells are sectioned transversely the larger aggregates of glycogen particles follow a curved course. Since outer hair cells of the apical region of the cochlea contain a bundle of contractile microfilaments, our results suggest that glycogen is associated with the contractile apparatus of these cells. This hypothesis is in good accordance with previous experimental data which suggest that glycogen is used as energy source for the contractile movements of outer hair cells.

Aging↗
PubMed1988

Neuron-specific enolase during the development of the organ of Corti.

NSE immunoreactivity has been studied in the organ of Corti of the developing mouse from birth to 21 days. NSE immunohistochemical stain is observed in spiral ganglion cells, in nerve fibers and in nerve endings of inner and outer hair cells, and in both populations of sensory cells. Spiral ganglion cells in lower and central parts of the ganglion stain for NSE at birth, but all nerve cells are stained by day 4. Radial and spiral fibers and the endings on inner hair cells stain at birth, but the nerve endings on outer hair cells develop NSE between days 3 and 6. The inner and outer hair cells are NSE-positive at day 2 but the NSE immunoreactivity in the outer hair cells decreases at the end of the second week until the cells become negative. The NSE stain in the neuronal pathways of the inner and outer hair cell regions increases for about 19 days, showing a predominant accumulation in neuronal endings. The data suggest that the development of NSE expression in the organ of Corti reflects the nascence and maturation of the synaptic contacts. Spiral neurons, their fibers and endings as well as inner and outer hair cells express NSE in the isolated organ of Corti in culture. Variability of stain among the different cell populations indicates a role of local factors in the regulation of NSE expression.

Animals↗
PubMedDate not supplied

Age-related degeneration of the organ of Corti in two genotypes of mice.

Degenerative changes in the organ of Corti were investigated in two genotypes of mice, CBA/Ca (moderate age-related late-onset hearing loss) and C57BL/6J (spontaneous early-onset auditory deterioration), ranging in age from 1 to 21 months (C57BL) and from 1 to 33 months (CBA). Light, transmission and scanning electron microscopies were used for qualitative and quantitative analyses. Alterations of deterioration in the CBA-mouse cochlea showed a late-onset loss of hair cells, lipofuscin in the outer hair cells (OHCs), giant stereocilia on the inner hair cells and elevated pillar heads. In C57BL mice, the primary lesions of the organ of Corti include early damage of stereocilia on OHCs, missing OHCs, and a ruptured reticular lamina. Giant cilia and elevated pillar heads were not evident in C57BL mice. In the aged C57BL mouse cochlea, the whole organ of Corti was collapsed and replaced by the supporting cells, whilst in the oldest CBA mice, completely normal-looking OHCs still remained even in severely deteriorated areas of the organ of Corti. The results indicate that the patterns of degeneration of the organ of Corti are different between CBA/Ca and C57BL/6J mice, probably under the control of different genes.

Aging↗