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PubMed1988

Immunoelectron microscopy identifies several types of GABA-containing efferent synapses in the guinea-pig organ of Corti.

Using an immunoperoxidase technique, we have localized by light and electron microscopy GABA-immunostained fibers within a component of the efferent innervation of the organ of Corti. At the light microscopic level, GABA-immunostained fibers were observed within the inner spiral bundle (below the inner hair cells) and the tunnel spiral bundle. The immunostaining was clearly more intense in the upper turns than in the basal turns. Mostly in the upper turns, GABA-immunostained fibers were seen crossing the tunnel of Corti to reach the outer hair cells where they formed large immunostained patches at the base of the cells. Unevenly distributed throughout these upper turns, immunostained fibers were seen climbing along the outer hair cells and traveling near the non-sensorineural Hensen's cells. The electron microscopic observations of GABA-immunostained fibers in the upper turns allowed us to identify within the inner spiral bundle vesiculated varicosities synapsing with radial dendrites connected to the inner hair cells. In the outer hair cell area, the GABA-immunostained fibers made several kinds of synaptic contacts. They included a minor population of the large axosomatic synapses with the basal pole of the outer hair cells and many axodendritic synapses with the spiral dendrites connected to these cells. Occasionally, the GABA-immunostained climbing fibers also synapsed with the outer hair cells at a supranuclear level. These result confirm previous light microscopic data dealing with the projection of the GABA-immunostained fibers along the cochlear partition. Moreover, they extend them in characterizing several kinds of GABA-immunostained synapses. These latter findings agree with previous neurochemical electrophysiological data which suggests an efferent neurotransmitter role for GABA. Nevertheless, such an existence of an efferent innervation predominantly projecting to the upper turns of the cochlea adds another criterion distinguishing the "apical" from the "basal" cochlea.

Animals↗
PubMed2005

Phase of neural excitation relative to basilar membrane motion in the organ of Corti: theoretical considerations.

Although the auditory transduction process is dependent on neural excitation of the auditory nerve in relation to motion of the basilar membrane (BM) in the organ of Corti (OC), specifics of this process are unclear. In this study, therefore, an attempt was made to estimate the phase of the neural excitation relative to the BM motion using a finite-element model of the OC at the basal turn of the gerbil, including the fluid-structure interaction with the lymph fluid. It was found that neural excitation occurs when the BM exhibits a maximum velocity toward the scala vestibuli at 10 Hz and shows a phase delay relative to the BM motion with increasing frequency up to 800 Hz. It then shows a phase advance until the frequency reaches 2 kHz. From 2 kHz, neural excitation again shows a phase delay with increasing frequency. From 800 Hz up to 2 kHz, the phase advances because the dominant force exerted on the hair bundle shifts from a velocity-dependent Couette flow-induced force to a displacement-dependent force induced by the pressure difference. The phase delay that occurs from 2 kHz is caused by the resonance process of the hair bundle of the IHC.

Acoustic Stimulation↗
PubMed1994

Expression of BDNF and NT-3 mRNA in hair cells of the organ of Corti: quantitative analysis in developing rats.

Brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) are synthesized by inner and outer hair cells of the developing organ of Corti. This raises the possibility that the reorganization of cochlear innervation patterns that occurs postnatally may be influenced by changing levels of neurotrophin expression. To determine if differential expression of BDNF or NT-3 in the inner and outer hair cells correlates with the reorganization of afferent and efferent innervation, we used in situ hybridization techniques to quantify relative levels of transcript biosynthesis in hair cells of developing rats. BDNF transcripts decreased in inner and outer hair cells from E17 to insignificant levels at P4. NT-3 expression was high at E17 in inner and outer hair cells, decreased in outer hair cells by E21, in inner hair cells by P1, remained low during the first postnatal week and was increased in the adult. The decreases in expression of both neurotrophins at birth precede the retraction of afferent nerve terminals from outer hair cells. BDNF and NT-3 transcription decreases substantially in outer hair cells between E21 and P4 when efferent innervation begins, indicating target biosynthesis of these neurotrophins is not likely to be instrumental in efferent target selection.

Animals↗
PubMed1994

Supranuclear efferent synapses on outer hair cells and Deiters' cells in the human organ of Corti.

Synaptophysin immunoreactivity and transmission electron microscopy have demonstrated vesiculated nerve endings synapsing on the supranuclear zone of outer hair cells and also of Deiters' cells in the human organ of Corti. These fibers seem similar to supranuclear fibers, apparently derived from the olivocochlear efferent system, which have been described in the animal. However, these endings were found throughout the cochlea in the human whereas in the animal such fibers were limited to the apical cochlea. Although such fibers have been demonstrated among supporting cells by immunohistochemical techniques, this is the first demonstration by transmission electron microscopy of morphology consistent with a chemical synapse between such fibers and Deiters' cells. Although the role of such fibers is unknown, neurophysiologic evidence suggests that they may modify the micromechanics of the outer hair cell. The function of neural innervation of Deiters' cells is speculative.

Hair Cells, Auditory, Outer↗
PubMed1997

Low-frequency modulation of inner hair cell and organ of Corti responses in the guinea pig cochlea.

Low-frequency tones are used to study changes in responsiveness as a function of phase in inner hair cell (IHC) and organ of Corti (OC) responses recorded from second turn of the guinea pig cochlea. In these experiments a 40 Hz stimulus is combined with a variable frequency probe to determine the degree to which tones at and below best frequency (BF) are modulated. Changes in responsiveness produced by the low-frequency input are quantified and related to position of the basilar membrane which is estimated using the phase of the cochlear microphonic measured in the OC fluid space. Results obtained when 40 Hz is presented at its lowest effective level demonstrate that ac responses to low-level BF probes are reduced for basilar membrane displacements to scala tympani while probe tones well below BF are modulated in the opposite direction. The transition between these two response patterns occurs when the overall DC produced in the OC by the two-tone input changes from positive to negative. Because of this association, the frequency dependence exhibited in the bias results may be linked to mechanisms responsible for generating the two polarities of the summating potential and the DC receptor potentials that it reflects. An attempt is also made to relate bias-induced changes in hair cell receptor potentials to modulations in single-unit rate responses. In other words, to address variations in the temporal relationships between excitation and suppression measured in the auditory nerve.

Acoustic Stimulation↗
PubMed2005

Endothelial nitric oxide synthase upregulation in the guinea pig organ of Corti after acute noise trauma.

Endothelial nitric oxide synthase (eNOS) upregulation was identified 60 h after acute noise trauma in morphologically intact cells of the reticular lamina in the organ of Corti of the guinea pig in the second turn of the cochlea. Using gold-coupled anti-eNOS antibodies and electron microscopy, it was shown that eNOS expression was upregulated in all cell areas and cell types except inner hair cells. Furthermore, eNOS was found in the organelle-free cytoplasm and in mitochondria of various cell types. The density of eNOS in mitochondria was considerably higher compared with the surrounding cytoplasm. Since eNOS activity is regulated by calcium, the eNOS detection was combined with calcium precipitation, a method for visualizing intracellular Ca2+ distribution. After acute noise trauma, intracellular Ca2+ was increased in all cell types and cell areas except in outer hair cells. Comparing the distribution patterns of eNOS and calcium, significantly elevated levels (P < 0.0001) of eNOS were detected within a 100 nm radius near calcium precipitates in all cuticular structures as well as microtubule-rich regions and Deiters' cells near Hensen cells. The observed colocalization lends support to the postulated mechanism of eNOS activation by Ca2+. eNOS upregulation after acute noise trauma might therefore be part of an induced stress response. The eNOS upregulation in cell areas with numerous microtubule- and actin-rich structures is discussed with respect to possible cytoskeleton-dependent processes in eNOS regulation.

Acoustic Stimulation↗
PubMedDate not supplied

Gentamicin-induced cytotoxicity involves protein kinase C activation, glutathione extrusion and malondialdehyde production in an immortalized cell line from the organ of corti.

The objective of the present study was to investigate the biochemical mechanisms underlying gentamicin cytotoxicity in OC-k3 cells derived from an immortalized cell line developed from the organ of Corti of transgenic mice. Administration of 50 microM gentamicin significantly reduced cell proliferation and viability, as well as initiating morphological changes associated with apoptosis. Protein kinase C (PKC) alpha activity was increased in gentamicin-treated cells, peaking 15 min after dosing (+138.2%). This PKCalpha increase was followed by a rise of glutathione (GSH) efflux and a concomitant 29% decrease in intracellular GSH levels at 30 min. PKCalpha-specific inhibitors blocked these cytotoxic effects. Gentamicin also increased malondialdehyde levels, while N-acetylcysteine, GSH and ascorbic acid prevented gentamicin-induced cell death. These findings suggest that gentamicin cytotoxicity involves a production of intracellular reactive oxygen species and a concomitant PKC-dependent fall of intracellular scavenging abilities (GSH), events that together drive cells to undergo apoptosis.

Animals↗
PubMed1983

Fine structure of the sensory epithelium of guinea-pig organ of Corti: subsurface cisternae and lamellar bodies in the outer hair cells.

The fine structure of subsurface cisternae and lamellar bodies in the outer hair cells of the guinea-pig organ of Corti was studied with thin sections and freeze-fracture replicas. Subsurface cisternae in the outer hair cells consist of multilayers along the lateral plasma membrane of the cell. The outermost layer is a flattened cistern in the upper part of the supranuclear region, but comprises a series of tubules in the lower part. Deeper layers are fenestrated cisternae in which disc-like areas are found in the upper part of the supranuclear region. Lamellar bodies consist of concentric layers of fenestrated cisternae and are located in the apical cytoplasm beneath the cuticular plate. They are continuous with the subsurface cisternae. In the supranuclear cytoplasm, bulges of the subsurface cisternae and the lamellar bodies are found. Dilated cisternae are also present. Some dilated cisternae contain many small vesicles, which display acid phosphatase activity. The dilated cisternae are considered as forms of the bulges undergoing transformation into multivesicular bodies. The possible role of the lamellar bodies, and the origin and fate of the subsurface cisternae are discussed.

Animals↗
PubMed1992

Distribution of glutamate-like and glutamine-like immunoreactivities in the rat organ of Corti: a light microscopic and semiquantitative electron microscopic analysis with a note on the localization of aspartate.

The light- and electron microscopic localization of glutamate and glutamine in the rat organ of Corti was studied by means of antisera raised against the respective amino acids coupled to carrier proteins. The light microscopic analysis was performed in semithin sections treated according to the peroxidase-antiperoxidase procedure. The two amino acids were visualized in the same ultrathin sections by use of postembedding immunocytochemistry with two different gold particle sizes. The distribution of aspartate-like immunoreactivity was also recorded, but only at the light microscopic level. In the hair cells, the level of glutamate-like immunoreactivity was higher than that in supporting cells but lower than that in the presumed glutamatergic terminals of cerebellar parallel and mossy fibres. The latter types of terminal were sampled from ultrathin sections that had been incubated under the same conditions as the cochlear sections. Within the hair cells, gold particles signalling glutamate were enriched on mitochondria but not on clusters of synaptic vesicles. Glutamine-like immunoreactivity was present in hair cells as well as supporting cells. The glutamate/glutamine ratio, expressed as the ratio between the respective gold particle densities, was considerably lower for hair cells compared with the cerebellar excitatory terminals. No consistent difference was found between outer and inner hair cells in relation to the levels and subcellular distribution of glutamate and glutamine immunoreactivities. Aspartate-like immunoreactivity was accumulated in outer hair cells, with some labelling also of border cells and Böttcher cells. While the present study confirmed the presence of glutamate in hair cells and demonstrated that these cells are also endowed with the important glutamate precursor glutamine, it revealed notable differences between hair cells and presumed glutamatergic terminals in the CNS. These could reflect differences in the synthesis and compartmentation of transmitter glutamate. Methodological factors could also contribute. Alternatively, the differences could be interpreted to suggest that the hair cell transmitter is not glutamate, but a similar compound. Aspartate could be a candidate in the case of the outer hair cells.

Animals↗
PubMedDate not supplied

Hensen's cell cyst of the organ of Corti.

The temporal bone collection at the Massachusetts Eye and Ear Infirmary contains the bones of two subjects with bilaterally symmetrical cyst formations in the Hensen's cell areas of the organs of Corti. In both subjects the cysts are located in the 8-14-mm regions of the cochlear ducts. Both cochleas of one subject show pressure atrophy of the outer hair cells, demonstrating that the cysts may have the potential for producing high-tone hearing loss. While the genesis and contents of the cysts are unknown, it is postulated that they may represent the coalescence of lipid droplets extruded from Hensen's cells.

Aged↗
PubMed1990

Biochemical features of major organ of Corti proteins (OCP-I and OCP-II) including partial amino acid sequence.

Further biochemical and biophysical characterization of two low-molecular-weight, strongly acidic proteins that are present at extremely high levels in the organ of Corti, tentatively named OCP-I and OCP-II, is presented. The two proteins are also present, although at much lower levels, in the vestibular end-organs and a variety of other inner ear tissues; they have not been observed in other systems. OCP-I and II are highly soluble and do not contain appreciable amounts of carbohydrate. The two proteins, originally described in the guinea pig, are compared electrophoretically with the corresponding proteins in several other mammalian species. Preliminary data on the amino acid composition of the two proteins are presented. Moreover, the amino-terminal sequence of a 22-residue segment of OCP-II is shown and compared to the sequences of known proteins.

Amino Acid Sequence↗
PubMed1987

Regional specialization of the cell coat in the hair cells of the organ of Corti.

The anionic sulfated groups of the cell coat glycoconjugates were studied in sensory and supporting cells of the organ of Corti with the colloidal iron hydroxide technique of Mowry, modified by Seno et al. [(1985) Histochemistry 82, 307-312]. This technique specifically stains the apical (endolymphatic) surface of both inner and outer hair cells. As this is the area at which the influx of potassium into the sensory cells takes place, the accumulation of negative charges in this location may be a mechanism to concentrate cations (mainly potassium) close to this portion of the hair cell membrane.

Animals↗
PubMed1994

Photo-released intracellular Ca2+ evokes reversible mechanical responses in supporting cells of the guinea-pig organ of Corti.

Loaded under whole cell patch-clamp configuration, the caged Ca2+ molecule DM-nitrophen was used to increase [Ca2+]i rapidly and reversibly in isolated Deiters cells of the organ of Corti. Photolysis of DM-nitrophen increased [Ca2+]i from resting concentrations of 20-50 nM to values above microM, as measured with the fluorescent indicator Fluo-3. Immediately after the photoliberation of Ca2+, a movement of the head of the phalangeal process could be observed in 75% of cells (n = 28). This mechanical movement, with an amplitude ranging between 0.5 to 1 micron within few hundred of ms, consisted of an extension of the phalanges away from the cell body. Measurement of phalangeal stiffness in transversal flexion toward the cell body ranged between 15-440 pN/micron. Stiffness can increase by 28 to 51% after rising [Ca2+]i. The results suggest Ca2+ as a potential intracellular messenger for active mechanical responses in Deiters cells.

Acetates↗