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PubMedDate not supplied

A novel, simple organotypic culture method to study the organ of Corti from the neonatal gerbil.

An original, simple organotypic culture method was developed to grow the organ of Corti from the neonatal gerbil on the bottom of a Petri dish. In comparison with the commonly used Maximov slide assembly method, this method is easier, less time-consuming, and more economic. Our results in this study using fluorescent live/dead viability assay and fluorescein-conjugated antineurofilament antibodies show that the cultured organ of Corti and spiral ganglion cells not only survived for at least 14 days but also maintained their basic organization and normal development in vitro. Therefore, our method can serve as a reliable and easier alternative to the traditional techniques for studying the development as well as other physiological properties of the cultured organ of Corti.

Animals↗
PubMed1993

The developing organ of Corti contains retinoic acid and forms supernumerary hair cells in response to exogenous retinoic acid in culture.

The mammalian organ of Corti has one of the most highly ordered patterns of cells in any vertebrate sensory epithelium. A single row of inner hair cells and three or four rows of outer hair cells extend along its length. The factors that regulate the formation of this strict pattern are unknown. In order to determine whether retinoic acid plays a role during the development of the organ of Corti, exogenous retinoic acid was added to embryonic mouse cochleae in vitro. Exogenous retinoic acid significantly increased the number of cells that developed as hair cells and resulted in large regions of supernumerary hair cells and supporting cells containing two rows of inner hair cells and up to 11 rows of outer hair cells. The effects of retinoic acid were dependent on concentration and on the timing of its addition. Western blot analysis indicated that cellular retinoic acid binding protein (CRABP) was present in the sensory epithelium of the embryonic cochlea. The amount of CRABP apparently increased between embryonic day 14 and postnatal day 1, but CRABP was not detectable in sensory epithelia from adults. A retinoic acid reporter cell line was used to demonstrate that retinoic acid was also present in the developing organ of Corti between embryonic day 14 and postnatal day 1, and was also present in adult cochleae at least in the vicinity of the modiolus. These results suggest that retinoic acid is involved in the normal development of the organ of Corti and that the effect of retinoic acid may be to induce a population of prosensory cells to become competent to differentiate as hair cells and supporting cells.

Animals↗
PubMed1992

Actin-binding and microtubule-associated proteins in the organ of Corti.

Actin-binding and microtubule-associated proteins regulate microfilament and microtubule number, length, organization and location in cells. In freeze-dried preparations of the guinea pig cochlea, both actin and tubulin are found in the sensory and supporting cells of the organ of Corti. Fodrin (brain spectrin) co-localized with actin in the cuticular plates of both inner and outer hair cells and along the lateral wall of the outer hair cells. Alpha-actinin co-localized with actin in the cuticular plates of the hair cells and in the head and foot plates of the supporting cells. It was also found in the junctional regions between hair cells and supporting cells. Profilin co-localized with actin in the cuticular plates of the sensory hair cells. Myosin was detected only in the cuticular plates of the outer hair cells and in the supporting cells in the region facing endolymph. Gelsolin was found in the region of the nerve fibers. Tubulin is found in microtubules in all cells of the organ of Corti. In supporting cells, microtubules are bundled together with actin microfilaments and tropomyosin, as well as being present as individual microtubules arranged in networks. An intensely stained network of microtubules is found in both outer and inner sensory hair cells. The microtubules in the outer hair cells appear to course throughout the entire length of the cells, and based on their staining with antibodies to the tyrosinated form of tubulin they appear to be more dynamic structures than the microtubules in the supporting cells. The microtubule-associated protein MAP-2 is present only in outer hair cells within the organ of Corti and co-localizes with tubulin in these cells. No other MAPs (1,3,4,5) are present. Tau is found in the nerve fibers below both inner and outer hair cells and in the osseous spiral lamina. It is clear that the actin-binding and microtubule-associated proteins present in the cochlea co-localize with actin and tubulin and that they modulate microfilament and microtubule structure and function in a manner similar to that seen in other cell types. The location of some of these proteins in outer hair cells suggests a role for microfilaments and microtubules in outer hair cell motility.

Actinin↗
PubMed1992

Degeneration followed by partial regeneration of the organ of Corti in deafness (dn/dn) mice.

The deafness mouse is a Mendelian recessive mutant which never hears and has no stimulus-related receptor or neural auditory responses. From birth through 12 days after birth (DAB), the organ of Corti develops normally as seen with light microscopy, except that the space of Nuel does not fully develop. The inner and outer hair cells are degenerating between 12 and 24 DAB and are gone by 45 DAB. As the hair cells degenerate, other cells of the organ of Corti become less recognizable, appear to collapse, and lose their identities as differentiated cells. By 45 DAB, from base to apex, the organ of Corti is composed of a low, roughly cuboidal epithelium with no distinguishing cell types; a hint of a tunnel of Corti remains at the apex. In the basal turn, the organ of Corti remains in this degenerated state through at least 460 DAB (senility for these mice). In the apical organ of Corti, considerable regeneration occurs between 45 and 90 DAB. By 90 DAB the apical turn of the organ of Corti has readily identifiable inner and outer pillar cells, inner and outer supporting cells, Hensen's cells, and Claudius' cells. A tunnel of Corti and space of Nuel are also present in the apex but there are no hair cells. Mechanisms are not known for either the degeneration or the regeneration.

Animals↗
PubMed1999

Melatonin and other antioxidants prolong the postmortem activity of the outer hair cells of the organ of Corti: its relation to the type of death.

The outer hair cells of the organ of Corti transform sound into electrical signals, beginning the nervous auditive process. These cells produce acoustic emissions when working routinely, known as otoacoustic emissions. Otoacoustic emissions (OAEs) are recorded from the hearing duct through a probe which incorporates a sound source and a sensitive microphone. On the other hand, the cochlea produces oxygen-derived free radicals and nitric oxide, in addition, melatonin is present in the cochlea. The authors have studied the influence of melatonin or an antioxidant mixture (alpha-tocopherol acid succinate, ascorbic acid, glutathione, and N-acetylcysteine) on the postmortem activity of the outer hair cells of the organ of Corti of the rat, measuring distortion product otoacoustic emissions. Control rats showed postmortem distortion product otoacoustic emissions for about 2 min when sacrificed by decapitation, and for about 3 min when sacrificed by chloroform inhalation. Melatonin prolonged the postmortem activity 3.5 times when the animals were sacrificed by decapitation, and 7 times when animals were sacrificed by chloroform inhalation. Similar results were obtained with the antioxidant mixture. Results show that melatonin and other antioxidants have, in general, a protective role on the postmortem activity of the outer hair cells of the organ of Corti.

Animals↗
PubMed1990

Electron microscopic observation of calcitonin gene-related peptide-like immunoreactivity in the organ of Corti of the rat.

Calcitonin gene-related peptide (CGRP)-like immunoreactive (CGRP-IR) nerve terminals in the organ of Corti of rats were studied by light and electron microscopy. Surface preparation of the organ of Corti were immunostained using anti-CGRP antiserum for avidin-biotin immunohistochemistry. Dense CGRP-IR fiber bundles were observed by light microscopy in the inner spiral bundles, tunnel spiral bundles and outer spiral bundles. Electron microscopic analysis indicated that CGRP-IR fibers belong to efferent nerves. In the inner spiral bundles, the CGRP-IR fibers showed a direct contact mainly with non-immunoreactive afferent fibers. Some CGRP-IR nerve endings in the inner spiral bundles formed contacts directly with inner hair cells. In the outer spiral bundles, CGRP-IR fibers formed synaptic contacts exclusively with the outer hair cells. It should be noted that the number of synapses of CGRP nerve endings with outer hair cells varied depending upon the sub-row: a falling gradient in number occurred along the inner-outer axis. Our results suggest that CGRP acts as an efferent neuromodulator in the organ of Corti.

Animals↗
PubMed1992

Postnatal development of the rat organ of Corti. I. General morphology, basilar membrane, tectorial membrane and border cells.

The development of the rat organ of Corti was studied during the first postnatal weeks. The temporal and the spatial patterns of cochlear development were investigated between 4 and 24 days after birth by means of semi-thin sections at approx. ten equidistant positions along the entire cochlear duct. At all examined positions width, thickness and cross sectional area of basilar membrane, cross-sectional area of tectorial membrane, of cells of Hensen, Claudius and Boettcher and of the organ of Corti were quantitatively analyzed. The most conspicuous maturational changes occur between 8 and 12 days after birth. These are the detachment of the tectorial membrane, the first appearance of filaments within the basilar membrane, the formation of the tunnel of Corti and the opening of the inner spiral sulcus. Quantitative analysis revealed that structures of a given position along the cochlear duct do not develop synchronously. Width of the basilar membrane and cross-sectional area of the tectorial membrane are already mature at the onset of hearing (10-12 days after birth). Length, thickness and cross-sectional area of the basilar membrane as well as cross-sectional area of the organ of Corti and of the cells of Hensen, Claudius and Boettcher still develop after the onset of hearing (up to 20-24 days after birth). We suggest that basic cochlear function is established by structures which are mature before the onset of hearing. Cochlear structures which develop after the onset of hearing might be involved in this improvement during this period.

Animals↗
PubMed1979

Intracellular potential changes of Corti's organ with anoxia.

Intracellular measurements of the resting cell membrane potentials of guinea pig Corti's organ were made in order to determine the sensitivity of this cell potential to anoxic hypoxia (a lowered oxygen state due to lack of respiratory oxygen) and to establish differences according to cell types or morphologic regions of the sensory epithelium. The negative cell potentials measured from successful electrode penetrations were found to be relatively more stable and resistant to change during a 120-s period of anoxia than was the positive endocochlear potential. The intracellular resting potentials were also much slower to recover after resumption of respiration. Data obtained from various cells in two different regions of Corti's organ is receiving oxygen from both the perilymph and the endolymph. An iontophoretic dye-marking technique was used to label some experimental cells for later histologic identification.

Action Potentials↗
PubMed2003

[Cisplatin-induced apoptotic cell death in spiral ganglion and organ of Corti of mongolian gerbil cochlear].

OBJECTIVE: To investigate whether apoptosis is one of the mechanism for cell death in spiral ganglion(SG) and organ of Corti of Mongolian gerbils cochlea induced by cisplatin. METHODS: Mongolian gerbils were administered 4 mg.kg-1.d-1 cisplatin consecutively for 4 to 7 days. The apoptotic cell death in spiral ganglion and organ of Corti of cochlea basal turn was detected via transmission electron microscopy and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) method. RESULTS: The observation of electron microscopy shown special morphological change consistent with the feature of cell apoptosis in some spiral ganglion cells and outer hair cells after 5-7 days' cisplatin treatment. The apoptotic labeling by TUNEL in spiral ganglion and organ of Corti was detected after 5 days' treatment and increased after 6 and 7 days' treatment whereas almost negative result were obtained in the normal control animals. CONCLUSION: Apoptosis is one of the important mechanisms for cell injury of spiral ganglion and organ of Corti induced by cisplatin.

Animals↗
PubMed1979

Uptake of putative neurotransmitters in the organ of Corti.

In vitro uptake of putative neurotransmitters into the organ of Corti of the guinea pig was studied by autoradiography. After incubation in 3H-glycine the label was heaviest over the inner hair cell, but was not confined to the synaptic region of the cell. After incubation in 3H-GABA, 3H-glutamate and 3H-aspartate, heavy labeling was seen over the fibers and terminals of the efferent olivocochlear bundle. Leucine, an amino acid not thought to be a neurotransmitter, was uniformly taken up by all cochlear structures. The fact that GABA, glutamate and aspartate are taken up into efferents, which are almost certainly cholinergic, suggests that high affinity uptake of these substances is not restricted to terminals in which these substances are released as neurotransmitters.

Animals↗
PubMed1991

The organ of corti, like the retina, should be considered as brain tissue.

The retina is considered as a part of brain tissue. The organ of corti, which is as specialized as the retina, is described as placodal in origin. A 14-point comparison--functional, evolutional, structural and biological--has been made between the organ of corti and the retina to justify the consideration of the organ of corti as also a part of brain tissue.

Animals↗
PubMed1997

Organization of cell junctions and cytoskeleton in the reticular lamina in normal and ototoxically damaged organ of Corti.

The reticular lamina creates an ion barrier, withstands mechanical stress in the organ of Corti and is able to maintain its integrity during and after severe hair cell loss. Tight junctions maintain the ionic gradient whereas adherens junctions and the cytoskeleton are responsible for the integrity and mechanical resistance of tissues. In this study we used immunofluorescence and electron microscopy to examine the distribution of proteins of tight junctions (cingulin), adherens junctions (E-cadherin, alpha- and beta-catenin) and the cytoskeleton (actin, cytokeratin and tubulin) in whole-mounts of the normal and ototoxically damaged organ of Corti. In normal ears the proteins of adherens junctions were found in all cell types of the reticular lamina. We now demonstrate that all cells forming the reticular lamina partially overlap each other organizing extensive cell contacts with a complex three-dimensional shape. During scar formation, the tight junctions as well as adherens junctions between hair and supporting cells appeared in two distinct focal planes, which could help to preserve the ionic barrier and tissue integrity during hair cell degeneration. During scar formation all cytoskeletal structures in the reticular lamina were reorganized in a specific spatio-temporal pattern. We present a three-dimensional model of cell contact organization in the reticular lamina of normal ears and during scar formation.

Actins↗
PubMed1993

Tissue culture of the organ of Corti.

In 1975, Sobkowicz et al. (1) described long-term organotypic cultures of the organ of Corti of the newborn mouse. This paper provides detailed methods for dissection and maintenance of the isolated organ of Corti with its corresponding segment of spiral ganglion in culture. Descriptions and illustrations of cellular characteristics of the developing organ are carefully documented. The work is based on 19 years of experience and over one thousand cultures. Review of the literature and the application of the technique to research on the inner ear are provided.

Animals↗
PubMed1986

The temperature dependence of electrical coupling in the organ of Corti.

Electrical coupling in an in vitro preparation of the organ of Corti was evaluated during changes in temperature of the bathing media. The effect of cooling the organ from 35 +/- 2 degrees C to 17 +/- 3 degrees C is to reduce membrane potentials, increase input resistance and decrease coupling ratios. Decreases in coupling ratios ranging from 15 to 75% have been observed. The effects are reversible upon warming. Membrane potentials are very susceptible to depolarization caused by cooling. The reduction in coupling is not due to depolarization nor is it dependent upon extracellular Ca2+. It is conceivable, however, that intracellular stores of Ca2+ are released or that intracellular pH is altered.

Animals↗
PubMed1986

Inhibition of myosin light-chain kinase activity in the organ of Corti by 0.3-5 kilodalton substances of the otosclerotic perilymph.

The guinea pig organ of Corti contains myosin light-chain kinase (MLCK) activity. The upper and lower most parts of the cochlea do not show significantly different activities of the enzyme, which is Ca2+ and calmodulin-dependent. Short-term noise exposure does not cause a significant change. 0.3-5 Kilodalton substances of the otosclerotic perilymph, separated by SG-25 column chromatography, inhibit the MLCK activity in in vitro organ of Corti preparations. This inhibitory action of the perilymph substances can also be observed with the purified MLCK of turkey gizzard. The activity of the enzyme can be specifically inhibited by cyclic AMP-dependent protein kinase.

Animals↗
PubMed1995

Ultrastructural localization of G-protein Gs in the organ of Corti.

Immunocytochemical localization of a stimulatory GTP-binding protein Gs in the organ of Corti in the inner ear was examined with a post-embedding immunogold technique, using antibodies raised against a synthetic decapeptide (RMHLRQYELL) of the C-terminus of the alpha subunit of Gs. Immunoreactivity was strong on the membranes of supporting cells in the reticular lamina, including inner and outer pillar cells and the phalangeal process of Deiters' cells. Immunolabeling also was seen on the membranes of cell bodies of those cells which surround nerve fibers, basilar fibers, outer spiral fibers and afferent nerve endings at outer hair cells. Gold particles also labeled the membrane of inner phalangeal cells and border cells. In contrast, outer and inner hair cells were not labeled. Possible roles of Gs in the organ of Corti are discussed.

Amino Acid Sequence↗
PubMed1986

Organ of Corti surface preparations for computer-assisted morphometry.

This paper describes a relatively rapid method (67 h) for producing surface preparations of the organ of Corti that are suitable for a computer-assisted morphometric analysis of cochlear hair cells. On day 1, a cochlea was fixed in OsO4, dehydrated in ethanol and infiltrated with an Epon-like plastic (Medcast). On day 2, the Medcast within the cochlea was polymerized at 60 degrees C in an oven. On day 3, the complete organ of Corti was dissected into approximately 20 segments which were trimmed and mounted on a slide in Medcast. Two 400 mesh transmission electron microscope grids are mounted at the apical and basal ends of the organ of Corti. These grids served as reference points for the establishment of an independent X, Y coordinate system on the slide by a computer-assisted light microscope. The segments were permanently attached to the slide by polymerization of the Medcast at 60 degrees C for approximately 15 h. At the beginning of day 4, the organ of Corti surface preparation was ready for examination by light microscopy or could be further sectioned and examined by transmission electron microscopy.

Animals↗
PubMedDate not supplied

Results of an ultrastructural study comparing stria vascularis with organ of Corti in guinea pigs treated with kanamycin.

Ultrastructural study of ototoxicity is well documented with two points of interest: organ of Corti for aminoglycosides and stria vascularis for loop diuretics. As a previous study suggested initial lesions of stria vascularis, an attempt of comparison and of chronological study was made between the organ of Corti and stria vascularis lesions by kanamycin intoxication. The method was devised by J. M. ARAN, with electrophysiological control. We failed to find in the stria vascularis a radial or longitudinal pattern of lesions. We could not discern a chronological injury between the organ of Corti and stria vascularis because both were damaged even in the less deafened animals. Nevertheless, two facts were clarified: hair cell lesions are lysosomial as for the kidney lesions, while stria vascularis lesions are mitochondrial, melanine granulations play a part in drug metabolism (increased number, secretory aspect) and deserve further study.

Animals↗