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Results for “Labyrinth Supporting Cells”

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At least 19 recordsLinked to original sources

PubMed2000

Characterization of an experimentally induced inner ear immune response.

OBJECTIVE: To determine the effects of a sterile immune response on the structure and function of the cochlea. METHODS: An immune response was created in guinea pigs by systemically sensitizing the animals to keyhole limpet hemocyanin and subsequently challenging the inner ear with the protein. Animals were allowed to survive for 1 to 5 weeks, after which the cochlea was evaluated histologically. Hearing was measured by auditory brainstem response before the inner ear challenge, during the survival period, and prior to sacrifice. RESULTS: Inflammatory cells infiltrated the cochlea from the circulation. Surface preparations and plastic sections of the organ of Corti 1 and 2 weeks after the initiation of the inflammation demonstrated degeneration of the sensory and supporting cells in cochlear turns containing inflammatory cells. Good preservation of structures was seen in the more apical cochlear turns with little or no inflammatory cells. In cochleas from animals that survived 5 weeks, most of the infiltrated cells were cleared after undergoing apoptosis and the inflammatory matrix in the scala tympani began to calcify. Hearing loss was moderate to severe depending on the amount of inflammation. CONCLUSION: Although in general the immune response serves to protect an organism from infection, these results demonstrate that bystander injury associated with local immune responses in the cochlea, an organ incapable of regeneration, causes permanent cochlear destruction and hearing loss.

Adjuvants, Immunologic↗

PubMed1997

Electrophysiological properties of vestibular sensory and supporting cells in the labyrinth slice before and during regeneration.

The whole cell patch-clamp technique in combination with the slice preparation was used to investigate the electrophysiological properties of pigeon semicircular canal sensory and supporting cells. These properties were also characterized in regenerating neuroepithelia of pigeons preinjected with streptomycin to kill the hair cells. Type II hair cells from each of the three semicircular canals showed similar, topographically related patterns of passive and active membrane properties. Hair cells located in the peripheral regions (zone I, near the planum semilunatum) had less negative resting potentials [0-current voltage in current-clamp mode (Vz) = -62.8 +/- 8.7 mV, mean +/- SD; n = 13] and smaller membrane capacitances (Cm = 5.0 +/- 0.9 pF, n = 14) than cells of the intermediate (zone II; Vz = -79.3 +/- 7.5 mV, n = 3; Cm = 5.9 +/- 1.2 pF, n = 4) and central (zone III; Vz = -68.0 +/- 9.6 mV, n = 17; Cm = 7.1 +/- 1.5 pF, n = 18) regions. In peripheral hair cells, ionic currents were dominated by a rapidly activating/inactivating outward K+ current, presumably an A-type K+ current (IKA). Little or no inwardly rectifying current was present in these cells. Conversely, ionic currents of central hair cells were dominated by a slowly activating/inactivating outward K+ current resembling a delayed rectifier K+ current (IKD). Moreover, an inward rectifying current at voltages negative to -80 mV was present in all central cells. This current was composed of two components: a slowly activating, noninactivating component (Ih), described in photoreceptors and saccular hair cells, and a faster-activating, partially inactivating component (IK1) also described in saccular hair cells in some species. Ih and IK1 were sometimes independently expressed by hair cells. Hair cells located in the intermediate region (zone II) had ionic currents more similar to those of central hair cells than peripheral hair cells. Outward currents in intermediate hair cells activated only slightly more quickly than those of the cells of the central region, but much more slowly than those of the peripheral cells. Additionally, intermediate hair cells, like central hair cells, always expressed an inward rectifying current. The regional distribution of outward rectifying potassium conductances resulted in macroscopic currents differing in peak-to-steady state ratio. We quantified this by measuring the peak (Gp) and steady-state (Gs) slope conductance in the linear region of the current-voltage relationship (-40 to 0 mV) for the hair cells located in the different zones. Gp/Gs average values (4.1 +/- 2.1, n = 15) from currents in peripheral hair cells were higher than those from intermediate hair cells (2.3 +/- 0.8, n = 4) and central hair cells(1.9 +/- 0.8, n = 21). The statistically significant differences (P < 0.001) in Gp/Gs ratios could be accounted for by KA channels being preferentially expressed in peripheral hair cells. Hair cell electrophysiological properties in animals pretreated with streptomycin were investigated at approximately 3 wk and approximately 9-10 wk post injection sequence (PIS). At 3 wk PIS, hair cells (all zones combined) had a statistically significantly (P < 0.001) lower Cm (4.6 +/- 1.1 pF, n = 24) and a statistically significantly (P < 0.01) lower Gp(48.4 +/- 20.8 nS, n = 26) than control animals (Cm = 6.2 +/- 1.6 pF, n = 36; Gp = 66 +/- 38.9 nS, n = 40). Regional differences in values of Vz, as well as the distribution of outward and inward rectifying currents, seen in control animals, were still obvious. But, differences in the relative contribution of the expression of the different ionic current components changed. This result could be explained by a relative decrease in IKA compared with IKD during that interval of regeneration, which was particularly evident in peripheral hair cells. (ABSTRACT TRUNCATED)

Animals↗

PubMedDate not supplied

Progenitor cell cycling during hair cell regeneration in the vestibular and auditory epithelia of the chick.

We investigated nucleotide-labeling patterns during ongoing hair cell regeneration in the avian vestibular epithelium and during drug-induced regeneration in the avian auditory epithelium. For utricle experiments, post-hatch chicks received an injection of bromodeoxyuridine (BrdU) and were allowed to survive from 2 hours to 110 days after the injection. Utricles were fixed and immunoreacted to detect BrdU. The number of BrdU-labeled nuclei in the hair cell and support cell layers of the utricular sensory epithelium changes significantly between 2 hours and 110 days post-BrdU. At 2 hours, most labeled cells are isolated, while by 5-10 days, the majority of labeled cells are organized in pairs that are most frequently composed of a hair cell and a support cell. Pairs of labeled cells are seen as late as 110 days. Clusters of more than 3 labeled cells are uncommon at all time-points. The total number of labeled cells increases approximately 1.5-fold between 5 and 60 days post-BrdU. This increase is due primarily to a rise in the number of labeled support cells, and it is likely that it represents additional rounds of division by a subset of cells that were labeled at the time of the BrdU injection. There is a significant decrease in labeled nuclei in the hair cell layer between 60 and 110 days post-BrdU, suggesting that hair cells die during this period. To investigate support cell recycling in the drug-damaged auditory epithelium, we examined nucleotide double labeling after separate injections of BrdU and tritiated thymidine. A small number of support cells that incorporate BrdU administered at 3 days post-gentamicin treatment also label with tritiated thymidine administered between 17 and 38 hours later. We conclude that a small population of support cells recycles during regeneration in both the normal utricle and the drug-damaged basilar papilla.

Animals↗

PubMedDate not supplied

Further observations of vestibular ototoxicity in the chick: effects of streptomycin on the ampullary sensory epithelium.

Starting eight days after hatching, chicks received daily subcutaneous injections of streptomycin sulfate, either 400 mg/kg for 30 days or 1,200 mg/kg for 15 days. Randomly selected chicks from each group were killed at intervals during the injection period, and the ampullae were examined for signs of vestibulotoxicity. Ampullary cell types differed in sensitivity to streptomycin. First, dark cell processes withered. Second, vacuoles formed in the apices of the light cells of the planum semilunatum. Third, nerve terminals swelled, and their organelles and ground substance clumped together, creating cleared areas. Last, hair cells and supporting cells became slightly vacuolated. However, these cells showed less overt damage than the other cell types. The onset of damage was earlier and the damage more severe with the 1,200-mg than with the 400-mg dosage. There was no evidence of hair cell loss during the experiment.

Animals↗

PubMed1993

Gap junctional connections between hair cells, supporting cells and nerves in a vestibular organ.

The pattern of gap-junctional connections between cells in the vestibular neuroepithelium of the posterior semicircular duct of the alligator lizard are described based upon the study of freeze fracture replicas and ultrathin sections with a transmission electron microscope. Both type I and type II hair cells are coupled to adjacent supporting cells by a series of small macular gap junctions located in a ring around the hair cell at the level of the apical circumferential belt of actin filaments. Adjacent supporting cells are extensively interconnected by gap junctions. A few cases of gap junctions between afferent dendrites and supporting cells, and between afferent dendrites and calyceal nerve endings were seen. These morphological observations together with data from other studies in the literature suggest a possible role for supporting cells in altering the micromechanical properties of the hair cell receptor organs during stimulation.

Actin Cytoskeleton↗

PubMed2006

MicroRNA gene expression in the mouse inner ear.

MicroRNAs (miRNAs) are small non-coding RNAs that function through the RNA interference (RNAi) pathway and post-transcriptionally regulate gene expression in eukaryotic organisms. While miRNAs are known to affect cellular proliferation, differentiation, and morphological development, neither their expression nor roles in mammalian inner ear development have been characterized. We have investigated the extent of miRNA expression at various time points throughout maturation of the postnatal mouse inner ear by microarray analysis. Approximately one third of known miRNAs are detected in the inner ear, and their expression persists to adulthood. Expression of such miRNAs is validated by quantitative PCR and northern blot analysis. Further analysis by in situ hybridization demonstrates that certain miRNAs exhibit cell-specific expression patterns in the mouse inner ear. Notably, we demonstrate that miRNAs previously associated with mechanosensory cells in zebrafish are also expressed in hair cells of the auditory and vestibular endorgans. Our results demonstrate that miRNA expression is abundant in the mammalian inner ear and that certain miRNAs are evolutionarily associated with mechanosensory cell development and/or function. The data suggest that miRNAs contribute substantially to genetic programs intrinsic to development and function of the mammalian inner ear and that specific miRNAs might influence formation of sensory epithelia from the primitive otic neuroepithelium.

Animals↗

PubMed1983

Distribution and polarity of actin in inner ear supporting cells.

Actin is present in the supporting cells of the chinchilla cochlea. Actin filaments which decorate with myosin subfragment S1 are found in both inner and outer pillar cells and in the Deiters' cells which surround outer hair cells. The opposing polarities of the S1 decorated actin filaments suggest that the supporting cells may play an active role in the structural support of the organ of Corti.

Actins↗

PubMedDate not supplied

Hair-cell numbers continue to increase in the utricular macula of the early posthatch chick.

It is generally assumed that hair-cell numbers do not increase in the vestibular epithelia of postembryonic birds after hatching. However, for the domestic chicken, it is not known when or if hair-cell numbers ever reach a steady state level during life. The numbers of hair cells in the utricular maculae of chickens from embryonic day (E) 7 to posthatch day (PH) 112 were therefore counted directly. Hair-cell numbers increase approximately 15 fold between E7 and PH2, from an average of 1,858/macula at E7 to 27,017 at PH2. Between PH2 and PH112 hair-cell numbers increase by a further 36%, to 36,650/macula. A mathematical description of the increase in hair-cell numbers observed with time predicts a half life of 29.88 days for a utricular hair cell and a steady-state turnover value of 850 hair cells/day by approximately PH60. The patterns of hair and supporting cells in the postembryonic utricular macula were also assessed. The ratios of supporting cells and hair cells, the average number of supporting cells around each hair cell, and the average number of hair cells each supporting cell contacts at PH2, PH16 and approximately 2.5 years of age are not significantly different. In contrast to the mitotically quiescent basilar papilla where all supporting cells contact at least one hair cell, 7.6% of supporting cells in the extrastriolar region of the postembryonic utricular macula do not make apical contact with a hair cell. These results indicate that hair-cell numbers in the utricular macula increase significantly after hatching, and support the concept that contact-mediated inhibition influences the proliferative potential of inner-ear supporting cells.

Acoustic Maculae↗

PubMedDate not supplied

Glutaraldehyde fixatives for preserving the chick's inner ear.

We preserved the inner ears of chicks in various concentrations of glutaraldehyde (2 to 3.5%) and cacodylate buffers (0.025 to 0.1 M). Buffer concentrations below 0.1 M caused osmotic damage that higher glutaraldehyde concentrations only partially counteracted. The combination of 3.5% glutaraldehyde and 0.1 M cacodylate buffer optimally preserved the different cell types and also eliminated problems of swelling and shrinkage. We further improved cellular preservation by immediately immersing the dissected specimen into chilled (4 degrees C), aerated fixative. The improved fixation greatly increased the retention of cytoplasmic ground substances, particularly in supporting cells and nerve terminals.

Aldehydes↗

PubMed1982

Supporting-cell and extracellular responses to acoustic clicks in the free-standing region of the alligator lizard cochlea.

1. Acoustic clocks were delivered to the tympanic membrane of anesthetized alligator lizards, and electric responses were measured within the free-standing region of the cochlea using glass micropipettes. Responses were recorded intracellularly in supporting cells and extracellularly in the receptor organ and in the scalae. Gross responses were also recorded with wire electrodes in scala tympani. 2. Intra- and extracellular responses contain two components: (1) an early (with latent period less than 0.15 ms after the onset of the click), rate-independent component presumed to originate in the receptor cells, which we call the 'receptor component'; and (2) a later (with latent period from 2 to 5 ms after the onset of the click), rate-dependent component presumed to originate in the primary neurons, which we call the 'neural component'. 3. The receptor component consists of a positive, slow, polarity-independent potential which is superimposed on a small, oscillatory, polarity-dependent potential. The average magnitude of the receptor component in supporting cells (0.72 mV for -20 dB clicks) is about 10 times that in the extracellular spaces and about 1/5 of that recorded in receptor cells. This component depends nonlinearly on the sound stimulus in the -20 to -55 dB range of click levels. 4. The average magnitude of the neural component in supporting cells (0.3 mV for clicks at -20 dB and 10 clicks/s) is about 5-10 times larger than that in the extracellular spaces. 5. The receptor and neural components have different distributions within the cochlea. The slow potential of the receptor component has positive polarity within the receptor organ and in scala tympani, and negative polarity in scala media. In contrast, the neural component has approximately the same biphasic (negative then positive) waveform in all extracellular compartments where it was detected. However, the neural component has a larger magnitude and an inverted (positive then negative) waveform in supporting cells. The neural component has not been detected in receptor cells.

Acoustic Stimulation↗

PubMed2006

Prospective identification and purification of hair cell and supporting cell progenitors from the embryonic cochlea.

Expression of the cyclin-dependent kinase inhibitor p27(Kip1) defines a post-mitotic population of cells in the embryonic mammalian cochlea that constitutes the nascent organ of Corti. Here, we describe techniques to purify these precursors using a transgenic p27/GFP reporter and fluorescence activated cell sorting (FACS). We demonstrate that these cells express other markers of the sensory lineage, such as Sox2, and when placed in dissociated cell culture differentiate as hair cells and supporting cells. The purified sensory progenitors thus obtained provide a means of studying the process of hair cell and supporting cell differentiation in vitro, as well as providing a means of analyzing the molecular and physiological properties of this unique population of cells.

Animals↗

PubMed1996

[Application of laser scanning confocal microscopy in inner ear morphological studies].

Laser scanning confocal microscope (LSCM) enables one to observe both the surface structure and the inner configuration in the same specimen, by its possibility of direct, non-invasive serial optical sectioning of whole mounted specimens. The potential value of LSCM in the field of inner ear morphological study was evaluated. The configuration of upper parts of organ of Corti was observed with the LSCM combined with double-stained fluorescence immunohistochemistry technique. The actin filament of hair cells by phalloidin, and the cytokeratin of supporting cells by monoclonal pan-anticytokeratin antibody. The stereocilia, cuticular plate, and the cuticle-free area of hair cells were well demonstrated. In the same specimen, the head plate of outer pillar cell, the phalangeal apical plate and the phalangeal process of Deiter cells were clearly showed as well. LSCM provide a new tool to the morphological study of organ of Corti.

Animals↗

PubMed1992

Atypical cells in the normal guinea pig organ of Corti as revealed by micromanipulation in SEM.

Inner ear tissue of the normal guinea pig was conductively stained (OTOTO-method) for SEM investigations. The Hensen's cells of the organ of Corti were removed using a micromanipulator inside the SEM. By this method atypical bodies of sensory and supporting cells were revealed in the apical turns of the cochlea. Atypical sensory cells showed great variations in size and shape. Several had no contact to Deiter's cells and no or only one nerve supply at their basal end. Atypical Deiter's cells showed alterations in shape and in the form of their phalangeal processes. Additionally altered parts of the organ of Corti were isolated by micromanipulation and embedded for correlative TEM-investigations.

Animals↗

PubMed1985

Outer hair cell loss and supporting cell expansion following chronic gentamicin treatment.

A sequence of changes in the organ of Corti associated with the destruction of outer hair cells (OHCs) and their replacement by supporting cells following chronic gentamicin treatment has been examined using thin-sections and SEM. The progression of change of OHCs was matched by concomitant expansion of adjacent supporting cells. Hair cells ruptured in the lateral membrane. The apical fragment was retained in the reticular lamina and became surrounded basally by the expanded supporting cells. No large breaches at the surface of the organ of Corti were formed. Rather, it appeared that the tight junctions around the hair cell were maintained until junctions were established between newly adjacent supporting cells in the space once occupied by the hair cell body. Only then was the OHC apex disrupted and the debris released into the sub-tectorial space. Some features of the OHC degeneration process were reminiscent of the controlled, cellular self-destruction phenomenon of apoptosis. The results suggest the possibility that the processes of hair cell loss and replacement may be controlled enabling maintenance of permeability barriers during structural reorganisation.

Animals↗

PubMed2002

Alteration in expression of p27 in auditory epithelia and neurons of mice during degeneration.

The aim of this study was to examine roles of p27, a cyclin-dependent kinase inhibitor, in cochleae of adult mice. Expression of p27 was found in cochlear supporting cells and spiral ganglion neurons of normal mice. Cisplatin treatment caused progressive degeneration of cochlear supporting cells and spiral ganglion neurons, and numbers of p27-positive cells in these cells decreased. This indicates a close relationship between p27 and cell death in cochleae. However, the relationships between decrease in number of p27-positive cells and that of survival cells differed according to type of cell. For Deiters' cells, there was apparent decrease in number of p27-positive cells, although no decrease in cell numbers. The present findings indicate that p27 plays roles in degeneration of cochleae according to cell type.

Animals↗

PubMed1985

Electron-microscopic observations of the gravity receptor epithelia of normal and spinner juvenile Octopus maya.

Light and electron microscopy of the gravity receptor epithelia (maculae) of statocysts of normal and "spinner" juvenile Octopus maya showed differences between the structures of the hair cells, supporting cells, and afferent neurons of these cephalopods. The maculae of spinner animals were approximately 30% smaller in their surface area and had 40% fewer hair cells. Moreover, the average distance between randomly-chosen hair bundles in scanning electron micrographs of maculae of normal animals was significantly greater (4.33 +/- 6.47 microns) than those of maculae of spinner animals (3.38 +/- 4.90 microns; P less than 0.0001). The sectional area of the supporting cell's microvilli in spinner maculae was larger (0.16 +/- 0.18 microns) than those of normal (0.10 +/- 0.10 micron; P less than 0.0001) O. maya. The morphological differences observed between certain structural components of the maculae of normal and spinner O. maya may be related to the absence and/or malformation of the neuroepithelial suprastructures in spinners. This may have direct or indirect effects to their inability to orient to gravity with these organs.

Acoustic Maculae↗

PubMed1993

The fine structure of the developing otolithic organs of the rat.

The fine structure of the otolithic organs in fetal and neonatal rats were studied by light and transmission electron microscopy. The otoconia were found at the 16.5 gestational day while the otolithic membrane appeared much later. Types II and I hair cells were first observed at the 16.5 and 18.5 gestational days, respectively. Secretory granules in the supporting cells seemed to release an organic material which was destined to be incorporated into the organic matrix of the otoconia and/or the otolithic membrane. Protrusions of apical cytoplasm, cytoplasmic globules and dilated rough endoplasmic reticulum (rER) were observed in epithelial cells of both sensory and non-sensory regions, in particular, in the transitional cells. By contrast, otoconia were mainly located above the neuroepithelial area. The functional roles of protrusions and cytoplasmic globules in the genesis of otoconia remain to be clarified. In addition, the pathway for transport of calcium remains obscure.

Animals↗

PubMedDate not supplied

Three-dimensional observation of the cochlea. Intracellular structure of the hair cell and the supporting cell.

Intracellular structures of the guinea pig cochlea were observed by scanning electron-microscopy. The cochlea was freeze-fractured and then macerated with 0.1% OsO4 solution (aldehyde-osmium-DMSO-osmium (AODO) method). Intracellular organelles, such as mitochondria, endoplasmic reticulum (ER), and Golgi apparatus, were demonstrated stereoscopically. The ER of the outer hair cell showed the most interesting features, such as subsurface cisternae and lamellar body. The subsurface cisternae which formed a stratiform network covered the inner surface of the cell membrane of the supranuclear part. Variously shaped mitochondria were found on the innermost layer of the subsurface cisternae. The lamellar body consisted of dilated cisternae and tubules of ER. The tubular ER of the lamellar body were contiguous with the subsurface cisternae. The pillar cells, Deiters' cells and Hensen's cells had well-developed tubular ER, while Claudius' cells had poorly developed tubular ER.

Animals↗