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PubMed1991

Isolated supporting cells from the organ of Corti: some whole cell electrical characteristics and estimates of gap junctional conductance.

Whole cell voltage clamp studies were performed upon isolated and small groups of supporting cells from the guinea pig organ of Corti in order to evaluate junctional and non-junctional membrane characteristics. Single Hensen cells have an average input resistance and capacitance of 1.03 G omega and 24.9 pF, respectively. I-V functions indicate an outward K+ rectification, which is blocked by external TEA, intracellular Cs, or photo-irradiation of intracellularly injected fluorescent dye. Voltage clamping of pairs or small groups of cells indicates that supporting cells 'share' K+ channels within the syncitium. The input impedance of coupled cells was studied during uncoupling with CO2 or octanol media. As expected, coupled cells showed an increase in input capacitance and a decrease in input resistance over single cell values. Input capacitance is a more sensitive indicator of cell coupling than dc input resistance. During uncoupling, input capacitance values drop to single cell levels prior to an increase of dc input resistance to single cell levels. Modeling the results indicates that Hensen cells are well-coupled under normal conditions and may have junctional resistances with values less than 0.1% of the non-junctional resistance. The sensitivity of the supporting cell syncitium's input impedance to small changes in junctional resistance markedly influences the syncitium's RC filter characteristics, and thus may control the frequency response of sound evoked electrical activity measurable in supporting cells in vivo.

Animals↗

PubMed1993

Partial amino acid sequences of organ of Corti proteins OCP1 and OCP2: a progress report.

Progress in amino acid sequencing of two low-molecular weight acidic proteins of unknown function which are present at high concentrations in the organ of Corti is reported. These two proteins, provisionally termed OCP1 and OCP2, were originally demonstrated by two-dimensional polyacrylamide gel electrophoresis; their presence at high concentrations in the inner ear sensory epithelia strongly suggests that OCP1 and OCP2 serve some important function in the ear. In the present paper, extension of the amino-terminal sequence of OCP2 to 71 residues is described. In addition, the first results on sequencing of OCP1 are presented. Computer algorithms are used to predict important structural features, and the sequences are analyzed for probable phosphorylation, glycosylation, and Ca-binding sites. Projected further studies in immunochemistry and molecular biology of OCP1 and OCP2 based on the amino acid sequencing data are discussed.

Amino Acid Sequence↗

PubMedDate not supplied

Comparative surface studies of ototoxic effects of various aminoglycoside antibiotics on the organ of Corti in the guinea pig. A scanning electron microscopic study.

It was the purpose of this study to establish criteria for use in comparing the toxic effects of aminoglycosid antibiotics on the organ of Corti by means of scanning electron microscopy. Amikacin, Tobramycin and Gentamicin were administered twice a day subcutaneously for 10 days to healthy guinea pigs. One group of animals was sacrificed 1 day after completion of the treatment; the other group was allowed to survive 22 days. Depending upon the dosage of the administered drug, Amikacin (150 mg per kg body weight daily, corresponding to 10 times an average recommended human dose) caused pronounced outer hair cell damage even 1 day after the treatment was stopped. At this time Gentamicin and Tobramycin (150 mg per kg body weight daily, corresponding to 50 times an average human dose) showed less damage. After 22 days' survival, late toxic effects were found mainly in Gentamicin- and Tobramycin-treated animals. After 3 weeks, nearly total outer hair cell loss was found in the basal coil, while the 2nd and 3rd coils were often less severely damaged. At this time Amikacin-treated animals showed severe damage in all coils. 300 mg per kg body weight Amikacin (i.e. 20 times the average human dose) showed about the same toxic effect on sensory cells of the guinea pig as did 150 mg Gentamicin or Tobramycin per kg body weight. We are conscious of the fact that there are problems in correlating the weight of a drug and its probable toxic effect. In comparative animal experiments we consider it useful to standardize the time of exposure, the amount of drug administered (e.g. related to the human dose) and the survival time.

Amikacin↗

PubMed1992

Localization of Ca(2+)-stores and tissue compartments with a Ca(2+)-binding capacity in the organ of Corti of the guinea-pig by electron energy-loss spectroscopy.

The addition of 10 mM CaCl2 to glutaraldehyde fixative leads to the formation of small electron-dense deposits in the organ of Corti of the guinea-pig. These precipitates are mainly attached to cell membranes in contact with different extracellular lymphatic fluids. A higher number of precipitates is localized in the acellular parts of tectorial and basilar membrane. Electron energy-loss spectroscopy (EELS) was used to determine the elemental composition of the deposits formed. The spectra showed a prominent signal at the Ca2+ L2,3 ionization edge. Oxygen could also be detected in all the precipitates analysed. EELS analysis of mitochondria of the inner and outer hair cells after conventional fixation (glutaraldehyde followed by post-fixation in OsO4) revealed a small but significant calcium signal.

Animals↗

PubMed1998

[Effect of the pulsed Er:YSGG and Ho:YAG laser on the organ of Corti of the guinea pig cochlear--a scanning electron microscopy study].

BACKGROUND: Recent experimental studies have demonstrated that, apart from the continuous wave lasers, several pulsed laser systems are also suitable for stapedotomy. The aim of this study was to clarify whether irradiation of the basal convolution of the guinea pig cochlea could cause morphological inner-ear changes using the Er:YSGG and Ho:YAG laser with laser parameters required for stapedotomy. METHODS: After opening the bulla, the basal convolution of the guinea pig cochlea, whose thickness is similar to that of the human stapes footplate, was irradiated with the Er:YSGG and Ho:YAG laser. The laser parameters used were those necessary for an adequate perforation of a human stapes footplate (500-600 microns). The cochleae were removed 90 minutes, 1 day, 2 weeks, or 4 weeks after laser application. The organ of Corti was examined by scanning electron microscopy in all convolutions. RESULTS: Application of Er:YSGG laser parameters effective for stapedotomy (5 pulses, energy: 85 J/pulse, energy density: 36 J/cm2, total energy: 0.425 J) had no adverse effects on the organ of Corti in the guinea pig cochlea. On the other hand, effective Ho:YAG laser parameters (10 pulses, energy: 210 J/pulse, energy density: 90 J/cm2, total energy: 2.1 J) caused damage to the outer hair cells with fusion of stereocilia and formation of giant cilia leading to partial or total cell loss. The inner hair cells and supporting cells were usually normal. CONCLUSION: Our results clearly demonstrate that the Er:YSGG laser has high application safety. It could prove to be a viable alternative to the thermically acting CO2 laser for stapedotomy. The Ho:YAG laser is not well tolerated in animals and has low application safety. Its use in stapedotomy would be unreliable and dangerous for the inner ear.

Animals↗

PubMed1999

Ionic currents expressed in a cell line derived from the organ of Corti of the Immortomouse.

We have investigated the maturation of adult hair cell electrophysiology in a population of precursor cells in a conditionally immortal cell line. The cell line, UB/OC-2, from the embryonic organ of Corti of the H-2Kb-tsA58 transgenic mouse, permits cells to grow proliferatively at 33 degrees C and to differentiate at 39 degrees C. Whole-cell patch-clamp recordings showed that proliferating cells had a different electrophysiology to differentiating cells. Differentiating cells had a conditionally expressed slowly activating inward current activated by hyperpolarization. The current was not blocked by extracellular application of 0.5 mM Ba2+, but was blocked reversibly by 2 mM Cs+. The current was found to be carried by both K+ and Na+ ions (PK/PNa=2.2) and activated by 10 microM forskolin. These properties identify the slowly activating current as Ih. A proportion of proliferating and differentiating cells exhibited a voltage-gated Na+ current, INa. INa was abolished in Na-free external medium and was inhibited reversibly by tetrodotoxin (TTX) with Ki=64 nM. Together these results suggest that proliferating and differentiating hair cell precursors in the immortal cochlear cell line UB/OC-2 express currents which are also found in developing hair cells.

Animals↗

PubMed1987

Succinate dehydrogenase and mitochondria in the hair cells in the organ of Corti of mature and old shaker-1 mice.

The activity of succinate dehydrogenase in the sensory hair cells in the organ of Corti of mature and old shaker-1 mice described as the animals grew older, the outer hair cells being the most effected. The decrease inactivity of this enzyme was closely linked with the degeneration of the mitochondria. When mitochondria were absent or grossly degenerated there was no enzyme activity. It is possible that this defect is linked with the deafness in these mice.

Animals↗

PubMed1990

Long-term effects of intense sound on hair cells of Corti's organ and endocochlear DC potential.

Cochleograms of guinea pig ears were made 30-40 days after exposure to intense pure tones of 300 Hz, 500 Hz, 2 kHz, or 4 kHz at 130-150 dB SPL for 4-24 h. At 4 kHz, hair cells in the basal turn disappeared totally, in the second turn moderately, and were relatively undamaged in the third and apical turns. At 500 Hz, hair cells in the second and third turns were almost completely injured and at 300 Hz moderately damaged in the third and apical turns although the basal turn remained undamaged. At 2 kHz for 9 h, hair cells were almost completely injured in all turns. Negative endocochlear DC potential (negative EP) induced by furosemide was observed in the basal turn but not in the third turn of animals exposed to 300 Hz. Contrarily, negative EP was observed in the third turn but not in the basal turn of animals exposed to 4 kHz. We conclude that the hair cells of Corti's organ play an essential role in the production of negative EP.

Acoustic Stimulation↗

PubMed1993

Three-dimensional cytoskeletal structures of the chinchilla organ of Corti: scanning electron microscopy application of the polyethylene glycol method.

We describe the application of a polyethylene glycol (PEG) embedding technique to examine the sensory and supporting structures of the inner ear. The chinchilla organ of Corti was exposed by cracking PEG embedded cochleas. A range of PEG molecular weights (2000-8000) were utilized; PEG 2000, with a melting point of 57 degrees C was preferred. After removal of the PEG, the three-dimensional aspects of intracellular structures were observed using scanning electron microscopy. Filamentous elements in the hair cell cuticular plate and in the supporting cells were clearly observed, as was the meshwork of cross-linked actin filaments in the cuticular portion of sensory hair cells. Microtubule and microfilament alignment patterns in pillar and Deiters cells were also clearly demonstrated. Characteristic structures in the outer hair cell synaptic region, such as the post-synaptic cistern and synaptic body, were well preserved using the PEG method.

Actin Cytoskeleton↗

PubMed1990

Can shape deformations of the organ of Corti influence the travelling wave in the cochlea?

Via their motile reactions outer hair cells may produce oscillatory power or amplify the fluid waves in the cochlea. When the cells move or change their lengths, the organ of Corti (OC) will change its shape. This paper describes the consequences of such shape changes for the physics of cochlear waves. The assertions posed are based on a mathematical derivation but the major conclusions can be grasped without following the mathematics in detail. There are two basic types of OC deformations. In the first type the net cross-sectional area of the OC is periodically varying, in the second type--the central subject of this paper--is is not. It is shown that for the latter type of OC shape deformation there is no interaction with the cochlear fluids, at least for long waves. Hence, in this case the outer hair cells cannot amplify cochlear waves. The other type of OC deformation has a better coupling with the cochlear waves. However, much of the pressure developed by oscillating hair cells is spent in the 'wrong' way, namely, by squeezing fluid lengthwise through the narrow channel of the OC. Therefore, in this mode power transfer from cochlear hair cells to cochlear fluids (and from there to the basilar membrane) would be very inefficient.

Animals↗

PubMed2003

Localization of the calcium channel subunits Cav1.2 (alpha1C) and Cav2.3 (alpha1E) in the mouse organ of Corti.

Voltage-activated Ca2+ channels play an important role in synaptic transmission, signal processing and development. The immunohistochemical localization of Cav1.2 (alpha1C) and Cav2.3 (alpha1E) Ca2+ channels was studied in the developing and adult mouse organ of Corti using subunit-specific antibodies and fluorescent secondary antibodies with cochlear cryosections. Cav1.2 immunoreactivity has been detected from postnatal day 14 (P14) onwards at the synapses between cholinergic medial efferents and outer hair cells as revealed by co-staining with anti-synaptophysin and anti-choline acetyltransferase. Most likely the Cav1.2 immunoreactivity was located presynaptically at the site of contact of the efferent bouton with the outer hair cell which suggests a role for class C L-type Ca2+ channels in synaptic transmission of the medial efferent system. The localization of the second Ca2+ channel tested, Cav2.3, showed a pronounced change during cochlear development. From P2 until P10, Cav2.3 immunoreactivity was found in the outer spiral bundle followed by the inner spiral bundle, efferent endings and by medial efferent fibers. Around P14, Cav2.3 immunoreactivity disappeared from these structures and from P19 onwards it was observed in the basal poles of the outer hair cell membranes.

Animals↗

PubMed2006

A PI3K pathway mediates hair cell survival and opposes gentamicin toxicity in neonatal rat organ of Corti.

Gentamicin is well known to promote hair cell death in inner ear, but it also appears to activate opposing pathways that promote hair cell survival. In combination with others, our previous work has indicated that a K-Ras/Rac/JNK pathway is important for hair cell death and an H-Ras/Raf/MEK/Erk pathway is involved in promoting hair cell survival (Battaglia et al., Neuroscience 122(4):1025-1035, 2003). However, these data also suggested that a Ras-independent survival pathway for activation of MEK might be stimulated by gentamicin. To investigate alternatives to the Ras/Raf/MEK/Erk pathway in promoting hair cell survival, cochlear explants were exposed to gentamicin combined with several inhibitors of alternative pathways (LY294002, calphostin C, SH-6, U73122). When exposed to gentamicin with the PI3K inhibitor LY294002 (10, 50 microM), the protein kinase C (PKC) inhibitor calphostin C (50, 100 nM) or the PKB/Akt inhibitor SH-6 (5, 10 microM), hair cell damage was significantly increased compared to gentamicin alone. By Western blotting, strong PKB/Akt activation was observed in the organ of Corti following exposure to 50 microM gentamicin for 6 h. In addition, PKC activation by 12-O-tetradecanoylphorbol-13-acetate protected outer hair cells from gentamicin induced cell death. In contrast, the phospholipase C-gamma (PLCgamma) inhibitor U73122 (2, 5 microM) did not affect hair cell damage when combined with gentamicin. Also, phosphorylation of PLCgamma was not increased in the organ of Corti following gentamicin treatment, as evaluated by Western blot. The results indicate that PI3K promotes hair cell survival via its downstream targets, PKC and PKB/Akt. This suggests that both Ras-dependent and Ras-independent survival pathways are involved during gentamicin exposure. In contrast, PLCgamma activation of PKC does not appear to play a role.

Animals↗

PubMed1991

Gentamicin-induced alterations of succinic dehydrogenase activity in the organ of Corti as revealed by non-decalcified frozen sections of the guinea pig's cochlea.

The distribution of succinic dehydrogenase (SDH) activity and the changes of SDH activity after injection of gentamicin (GM) were observed in the organ of Corti using non-decalcified frozen sections of the guinea pig's cochlea. The distribution of SDH activity was found to increase from the apex to the basal turn. At each turn, SDH activity of the inner hair cells, the inner supporting cells and the nerve endings surrounding the supporting cell and on the hair cells presented a greater activity than that found in the outer hair cells, adjacent Deiter's cells and associated nerve endings. It was further observed that GM had a greater effect on SDH activity in the basal turn than the other turns. At each turn, a more sensitive area of response to GM was found on the nerve endings one each hair cell, especially on the outer hair cells of the basal turn.

Animals↗

PubMed1993

Characterization of Ca2+ signals generated by extracellular nucleotides in supporting cells of the organ of Corti.

ATP has been demonstrated to act as a co-transmitter or neuromodulator in various physiological processes. There is recent evidence that ATP receptors, characterized as P2 purinergic receptors, are expressed in the sensory hair cells of the auditory organ. The aim of the present study was to know whether other cell types of the organ of Corti, the supporting cells, were also sensitive to external ATP. In both types of supporting cells considered in this study, Deiters' cells (DCs) and Hensen's cells (HEs), extracellular ATP at sub-micromolar concentrations evoked a transient increase in [Ca2+]i as monitored with fluorescence microscopy using the calcium probe Indo-1. An apparent Kd of 0.5 and 0.9 microM was determined for DCs and HEs, respectively. Furthermore, we demonstrated that ATP stimulated Ca2+ release from internal stores in DCs, but not in HEs. Dynamic calcium imaging by confocal laser scanning microscopy of ATP induced Ca2+ mobilization demonstrated a calcium wave propagation in the cell body of DCs which originated in the phalangeal processes, suggesting a functional organization of Ca2+ sequestering stores in DCs.

Adenosine Triphosphate↗

PubMed1986

Intracellular recordings from supporting cells in the organ of Corti.

Horseradish peroxidase marking and intracellular recording techniques were utilized to examine whether the heterogeneity of supporting cell types is reflected in their electrical behavior. The magnitudes of ac and dc response components were compared to their extracellular counterparts. The relationships among these potentials seem to be governed by the cell's location within the organ of Corti.

Animals↗

PubMed2001

Intracellular calcium dynamics and membrane conductance changes evoked by Deiters' cell purinoceptor activation in the organ of Corti.

Deiters' cells function as supporting cells for the sensory-motor outer hair cells of the mammalian cochlea and are interconnected by gap junctions. Here the electrical and Ca2+ responses of Deiters' cells evoked by purinergic stimulation were investigated in the organ of Corti, the auditory sensory epithelium. Adenosine 59-triphosphate (ATP, 50-100 microM) applied focally by pressure increased the intracellular free Ca2+ concentration ([Ca2+]i). At the same time ATP evoked an early inward current that was followed by an outward component, reflecting a sustained Ca2+-dependent reduction of the pre-stimulus offset current. These responses were maintained when Ca2+ was removed from the extracellular medium (0 [Ca2+]o), indicating a contribution to Ca2+ signalling from P2Y metabotropic receptors. UV photolysis of caged inositol 1,4,5-triphosphate (InsP3, 16 microM) produced Ca2+ responses similar to those evoked by exogenous ATP, accompanied by reduction of the offset current. In Deiters' cells uncoupled by octanol (1mM), ATP activated only the early inward current, suggesting that functional gap junctions are required in the late phase of the current responses. Following the delivery of UV flashes to pairs of Deiters' cells loaded with caged InsP3, the electrical coupling ratio (CR), monitored by double patch-clamp recordings, was strongly attenuated. These data support the idea that, by promoting inflow of cations and by controlling gap-junction conductance in a Ca2+-and InsP3-dependent way, ATP might serve a protective role in the cochlea.

Animals↗

PubMed1979

Structural changes in the organ of Corti of the guinea pig after obstruction of the arterial blood flow to the inner ear.

Forty-one young guinea pigs were studied after obstruction of the anterior inferior cerebellar artery in order to ascertain the effects on the surface specimen of the organ of Corti. Most of the animals had damage in the third and fourth turns (22) and a minority of these had dizziness and destruction nystagmus (3). Our hypothesis is that one of the causes of Meniere's disease is an interference with the inner ear circulation, and our plans for testing this hypothesis are given.

Animals↗

PubMed2005

Recombinant human erythropoietin prevents ischemia-induced apoptosis and necrosis in explant cultures of the rat organ of Corti.

This study was designed to evaluate the effect of recombinant human erythropoietin (rhEPO), insulin-like growth factor-1 (rhIGF-1) and epidermal growth factor (rhEGF) on ischemia-induced hair cell loss in an organotypic cochlea culture. The apical, middle and basal parts of the organs of Corti (newborn rat, postnatal days 3-5) were exposed to ischemia (3.5 h) in glucose-free artificial perilymph (pO2 10-20 mmHg) with or without growth factors. Controls were exposed to normoxia. Twenty-four hours after the onset of ischemia, the cultures were stained using tetramethyl rhodamine isothiocyanate (TRITC) phalloidin (hair cells), propidium iodide (membrane integrity) and apoptosis detection kit (DNA-fragmentation). Ischemia (3.5 h) induced a hair cell loss of 20 and 40% in the middle and basal cochlear parts, respectively, and an increase of the numbers of PI-stained and DNA-fragmented nuclei (controls 0-1, ischemia 4-7 nuclei/100 microm). The basal part was more affected than the apical one. rhEPO and rhIGF-1 significantly attenuated the ischemia-induced hair cell loss by reducing processes involved in apoptosis and necrosis. rhEPO has been in clinical use for more than a decade and found to be well tolerated. Therefore, rhEPO could be an effective drug for the prevention of hearing loss via a hair cell protective mechanism.

Animals↗