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PubMed2003

Comparative expression patterns of T-, N-, E-cadherins, beta-catenin, and polysialic acid neural cell adhesion molecule in rat cochlea during development: implications for the nature of Kölliker's organ.

We investigated the expression patterns of several cell adhesion molecules (CAMs) during rat cochlea ontogeny, from embryo day 16 to adulthood, with the use of immunohistochemistry: neural cadherin (N-cad) and polysialic acid neural CAM (PSA-NCAM) as two different neural CAM paradigms; epithelial cadherin (E-cad), which was restricted to the epitheloid phenotype; and the cytoplasmic domain-free truncated-cadherin (T-cad). We made the following observations. (1) T-cad was present in all types of fibrocyte and in subdomains within the pillar cells. (2) E- and N-cad were expressed with mutually exclusive patterns and did not overlap with T-cad. All cochlear epithelial cells, including the sensory outer hair cells (OHCs), were E-cad-positive, except for the negative inner hair cells (IHCs) and the nonsensory Kölliker's organ domain close to the IHCs. N-cad expression appeared first in the developing IHCs and then in the neighboring Kölliker's organ in an increasingly mediolateral gradient in opposition to the E-cad gradient. The OHCs, which are never N-cad positive, intensively expressed E-cad, as did the Hensen cells at the beginning of their differentiation. (3) The cadherin-linked molecule beta-catenin, absent in fibrocytes, was detected in all epithelial cell membranes and was prominent in the E-cad-rich modiolar extremity of Kölliker's organ. (4) Gradual PSA-NCAM expression was observed in the lateral portion of Kölliker's organ, and the intense PSA-NCAM expression was seen surrounding the IHCs. As development proceeded, PSA-NCAM immunoreactivity progressively became restricted to the basal poles of the IHCs, where it remained in the adult rat cochlea, suggesting a synaptic plasticity. Synaptic plasticity in rat cochlea and hypotheses about T-cad functions and neosensory features of the Kölliker's organ are discussed.

Animals↗

PubMed1985

Projections from the cochlear nucleus to the inferior colliculus in normal and neonatally cochlea-ablated gerbils.

The distribution of the projection from one cochlear nucleus (CN) within each inferior colliculus (IC) was studied in adult, normal gerbils and adult gerbils subjected to unilateral ablation of the contralateral cochlea at 2 days of age. The projection was studied by using the Fink-Heimer technique for impregnating degenerating axons and their terminal processes with silver. Following an extensive, unilateral lesion of the CN, degeneration was seen in both ICs of all animals. In normal animals, degeneration was both more widespread and heavier in the contralateral than in the ipsilateral central nucleus of IC (ICC). Degeneration was most widespread in the rostral and lateral parts of both ICCs and in the ventral part of the contralateral ICC. Degeneration was observed in 26% of the area examined in ipsilateral ICC and in 73% of the area examined in contralateral ICC. In cochlea-ablated animals there was a much greater similarity in the area of degeneration in the ICC ipsilateral (57%) and contralateral (67%) to the CN lesion. The same regional distributions of degeneration were observed as in the normal animals except that the distribution of degeneration in the ipsilateral ICC more closely resembled the normal contralateral than the normal ipsilateral profile. We conclude that the normal distribution of projections from the CN within the ipsilateral ICC is substantially modified by neonatal ablation of the contralateral cochlea.

Animals↗

PubMed1987

Selective retrograde labeling of lateral olivocochlear neurons in the brainstem based on preferential uptake of 3H-D-aspartic acid in the cochlea.

We have previously shown that perfusion of the gerbil cochlea with probe concentrations of 3H-D-aspartic acid (D-ASP) results in immediate, selective labeling of 50-60% of the efferent terminals under the inner hair cells, presumably by high-affinity uptake. The present study was undertaken to determine the origin of these endings. Twenty-four hours after cochlear perfusion with D-ASP, labeled neurons were observed in the ipsilateral, and to a much lesser extent in the contralateral, lateral superior olivary nucleus (LSO). The cells were small, primarily fusiform, and showed fewer synaptic contacts than other LSO cells. Combined transport of D-ASP and horseradish peroxidase indicated that all olivocochlear neurons within the LSO that projected to the injected cochlea were labeled by D-ASP. Labeled fibers coursed dorsally from the LSO, joined contralateral fibers that had passed under the floor of the fourth ventricle, and entered the VIIIth nerve root at its ventromedial edge. Adjacent to the ventral cochlear nucleus (VCN), densely labeled collateral fibers crossed the nerve root to enter the VCN. Labeled fibers and terminals were prominent in the central VCN. Neither retrograde transport of D-ASP by medial olivocochlear and vestibular efferents nor anterograde transport by VIIIth nerve afferents was observed. The D-ASP-labeled cells and fibers are clearly lateral olivocochlear efferents. Retrograde transport of D-ASP thus allows the cells, axons, and collaterals of the lateral olivocochlear system to be studied, morphologically, in isolation from other cells that project to the cochlea. Since the olivocochlear neurons are almost certainly cholinergic, retrograde amino acid transport does not necessarily identify the primary neurotransmitter of a neuron. Rather, it indicates the presence of selective uptake by the processes of that neuron at the site of amino acid injection. Retrograde labeling appears to be markedly enhanced by the use of metabolically inert compounds such as d-isomer amino acids.

Animals↗

PubMed1990

Auditory brainstem of the ferret: early cessation of developmental sensitivity of neurons in the cochlear nucleus to removal of the cochlea.

The role of primary afferent innervation in the maintenance of neurons in the mammalian auditory system was assessed by performing unilateral removals of the cochlea in neonatal and mature ferrets of known birth dates. Removals were performed under steroid anesthesia and resulted in the complete destruction of the organ of Corti and the loss of at least 80% of type I spiral ganglion neurons. Four main age groups [postnatal days (P)5, P24, P90, and P180] were used. Additional animals received no surgery, partial removals, or complete removals at older ages. Three months after the cochlear removal the animals were reanesthetized and perfused. The brainstem and the temporal bones were wax-embedded, frontally sectioned, and Nissl-stained. Sections of the right and left cochlear nuclei were compared quantitatively. Removal of the cochlea at P5 resulted in the loss of more than 50% of large (nongranular) neurons throughout the ipsilateral cochlear nucleus. Lesions at older ages did not produce any neuron loss. The size of the remaining neurons was reduced by 10-15% in all age groups. Partial lesions at P5 produced a graded response in the cochlear nucleus that was related to the extent of the lesion. The developmental sensitive period for the effects of cochlear removal on the ferret cochlear nucleus is therefore over before the age (P28-P30) at which the animal begins to hear. The present result differs markedly from the chicken, in which the sensitive period for removal of the cochlea persists for at least 2 months after the onset of hearing.

Afferent Pathways↗

PubMed1991

The sensory epithelium and its innervation in the mole rat cochlea.

The mole rat (Spalax ehrenbergi) burrows throughout its life in subterranean tunnels. Several structural and functional features which adapt the mole rat to its habitat have been discerned. The goal of this study was to elucidate the structural basis for adaptation of the auditory end-organ to an environment where low-frequency acoustical signals prevail. For this purpose, cochleae of adult mole rats were studied with light and electron microscopy. Inner hair cells throughout the cochlea, and outer hair cells in the basal (high-frequency) portion of the cochlea, were similar in structure to those seen in other mammals. In contrast, outer hair cells in the apical (low-frequency) portion displayed unique structural features. These features resembled the structure of inner hair cells or immature outer hair cells. The innervation of outer hair cells was most uncommon, in that classical medial efferent innervation was not found throughout the cochlear spiral.

Animals↗

PubMed1991

Hair cell differentiation in the developing chick cochlea and in embryonic cochlear organ culture.

We have defined a method for growing chick embryonic cochleae in organ culture that preserves many aspects of hair cell differentiation. Cochlear ducts were isolated from embryonic day 8 chicks, placed in organ culture, and incubated for 48 hours (to a point equivalent to embryonic day 10). The cultured ducts were then fixed and processed for scanning electron microscopy. As controls, cochlear ducts at embryonic days 8 and 10 were dissected and immediately fixed and processed for scanning electron microscopy. We chose this period to culture cochleae because at the corresponding time in vivo hair cells undergo a dynamic phase of differentiation. During this time, the number of stereocilia in the stereociliary bundle increases, and two to three rows of stereocilia nearest the kinocilium elongate, initiating the staircase pattern of the bundle. Also, the orientation of many hair cells shifts from nonpolarized at embryonic day 8 to polarized toward the inferior edge of the basilar papilla at embryonic day 10. Many of these aspects of hair cell differentiation proceed normally in organ culture. The appropriate distal-to-proximal gradients of hair cell density, apical surface area, and stereociliary number are preserved. Elongation of the 1-2 stereociliary rows next to the kinocilium continues, and more stereociliary bundles are oriented toward the inferior edge in cultured cochleae than in embryonic day 8 chicks. It appears that cochlear organ culture can serve as an effective method with which to study how hair cell differentiation is regulated.

Animals↗

PubMed1995

Distribution of non-NMDA glutamate receptor mRNAs in the developing rat cochlea.

In situ hybridization was used to document the distribution of mRNA encoding six subunit isoforms of non-N-methyl D-aspartic acid (NMDA) glutamate receptors (GluR1, GluR2, GluR3, GluR4, GluR5 and GluR6) in the inner ears of embryonic, postnatal and adult rats. GluR2 and GluR3 expression in the spiral ganglion appeared well before birth, and reached adult levels several days before the onset of function in the cochlea. In the spiral limbus, expression of GluR2 and GluR3 mRNA reached very high levels at around the time of birth, then declined after a few days. Low levels of GluR1, GluR4 and GluR6 expression were detected in various tissues of the cochlea during development. In the adult cochlea, GluR expression was limited to GluR2 and GluR3 mRNAs in the spiral ganglion neurons and GluR2 mRNA in fibrocytes of the spiral limbus, a non-neural tissue. The ontogenetic expression of additional GluR subunit genes and their appearance in different cochlear tissues could reflect different roles for these genes during development, or less precise regulation of gene expression within the GluR family. In particular, the very high levels of GluR gene expression in the spiral limbus during the perinatal period support a non-neural function, perhaps as cell surface receptors during tissue differentiation.

Animals↗

PubMed1990

Scanning electron microscopy of nerve fibers in human fetal cochlea.

The cochleas of four human fetuses ranging 22-25 weeks gestation were studied by scanning electron microscopy (SEM) for the purpose of obtaining a better understanding of the nerve fiber arrangement in the human ear. After critical point drying, the specimens were dissected and the floor of the tunnel of Corti and the outer wall of Nuel's space were exposed for observation. Upper cochlear turns, especially the apical turn, seemed to be still immature. Observed nerve fibers were classified into three types: 1. Spiral fibers: Fibers traveling basalward and following the shape of the cochlea were found in both the tunnel of Corti and Nuel's space and believed to be the afferent nerves responsible for innervating the outer hair cells. 2. Radial fibers: radiating outward from the osseous spiral lamina--one such radial fiber transversing high in the tunnel space (supposedly the efferent nerve servicing the outer hair cells), and another sort of radial fiber (found crossing the tunnel floor), the nature of which was uncertain 3. Irregular fibers: Consisting of thin, randomly running fibers within the cochlea. The destination of these fibers was not determined, but possibly they represent transitory nerve branchings of afferent or more probably efferent nerves, which would later regress during maturation.

Afferent Pathways↗

PubMed1990

Biochemical and morphological differentiation of acetylcholinesterase-positive efferent fibers in the mouse cochlea.

We have compared the biochemical expression of cholinergic enzymes with the morphological differentiation of efferent nerve fibers and endings in the cochlea of the postnatally developing mouse. Choline acetyltransferase (ChAT) and acetylcholinesterase (AChE) are present in the newborn cochlea at specific activities 63% and 25%, respectively, of their mature levels. The relative increases in ChAT, in AChE, and in its molecular forms over the newborn values start about day 4 and reach maturity by about day 10. The biochemical results correlate well with the massive presence of nerve fibers stained immunocytochemically for ChAT and AChE or enzymatically for AChE in the inner and outer hair cell regions. Ultrastructral studies, however, indicate the presence of only few vesiculated fibers and endings in the inner and outer hair cell regions. The appearance of large, cytologically mature endings occurs only toward the end of the third postnatal week. The discrepancy may be resolved in the electron microscopy using the enzymatic staining for AChE. Labeling is seen on many nonvesiculated fibers and endings in the hair cell regions, suggesting that the majority of the efferent fibers in the perinatal organ may be biochemically differentiated but morphologically immature. The results may imply that the efferents to inner and outer hair cells develop earlier than indicated by previous ultrastructral studies. Moreover, the pattern of development suggests that in the cochlea, as in other tissues, the biochemical differentiation of the efferent innervation may precede the morphological maturation.

Acetylcholinesterase↗

PubMed2005

Protein-protein interactions of a Kvbeta subunit in the cochlea.

Accessory subunits associated with voltage-gated potassium (Kv) channels can influence the biophysical properties and promote the surface expression of channel-forming alpha-subunits. Previously, we cloned several alpha-subunits and a beta-subunit from a cDNA library of the chicken cochlea. In the present study, we raised an antibody against the N-terminus of chicken Kvbeta1.1 (cKvbeta1.1) and characterized the Kvbeta-related polypeptide in cochlear tissues and heterologous cells. The anti-cKvbeta1.1 antibody recognizes a 45-kDa polypeptide in chick cochlear extracts as well as in Chinese hamster ovary (CHO) cells transfected with cKvbeta1.1. The accessory subunit was localized to the ganglion cells of the chick cochlea using immunohistochemistry and in situ hybridization. Coimmunoprecipitation studies show that Kvbeta1.1 interacts with Shaker channel members Kvalpha1.2 and 1.3, both of which colocalize with beta to the cochlear ganglion cells. Additionally, coimmunoprecipitation studies show that Kvalpha1.2 and 1.3 interact with each other, suggesting that these ion channels are formed by heteromultimers. In comparison, Kvbeta did not coprecipitate with a member of the Shal subfamily. The presence of Kvbeta in the cochlea suggests that this subunit contributes to the modulation of auditory signals in the ganglion cells, presumably by regulating properties of inactivation as well as surface expression of Kvalpha channels.

Animals↗

PubMed2005

Isolation and culture of hair cell progenitors from postnatal rat cochleae.

Cochlear hair cells are a terminally differentiated cell population that is crucial for hearing. Although recent work suggests that there are hair cell progenitors in postnatal mammalian cochleae, isolation and culture of pure hair cell progenitors from a well-defined cochlear area have not been reported. Here we present an experimental method that allows isolation and culture of hair cell progenitors from postnatal rat cochleae. These progenitor cells are isolated from the lesser epithelial ridge (LER, or outer spiral sulcus cell) area of pre-plated neonatal rat cochlear segments. They express the same markers as LER cells in vivo, including ZO1, Islet1, Hes1, and Hes5. When these cells are induced to express Hath1, they show the potential to differentiate into hair cell-like cells. Interestingly, these cells can be lifted from monolayer cultures and maintained in aggregate cultures in which spheres can be formed. Hair cell progenitors in the spheres display their proliferating capability and express only epithelial markers. Furthermore, when these spheres are mixed with dissociated mesenchymal cells prepared from postnatal rat utricular whole mounts, and replated onto a collagen substratum, the epithelial progenitor cells are able to differentiate into cells expressing markers of hair cells and supporting cells in epithelial islands, which mirrors the inner ear sensory epithelium in vivo. Successful isolation and culture of hair cell progenitors from the mammalian cochlea will facilitate studies on gene expression profiling and mechanism of differentiation/regeneration of hair cells, which are crucial for repairing hearing loss.

Actins↗

PubMed1999

Identification of aquaporin 5 (AQP5) within the cochlea: cDNA cloning and in situ localization.

Dysfunction of fluid and electrolyte homeostasis is considered to cause variety of inner ear disorders. One group of candidate proteins that may play a critical role in the inner ear fluid homeostasis is the aquaporins, a family of proteins whose members have well defined roles in fluid transport in variety of organs. This study reports the identification of AQP5, a member of the aquaporin family, within the rat inner ear and its in situ localization. AQP5 was initially identified within rat cochlear RNA via RT-PCR and sequence analysis of the amplified fragments. Immunoblot of cochlear homogenate yielded a predominant AQP5-immunoreactive band of M(r) 35 kDa. The anti-AQP5 immunoreactivity, indicating expression of the AQP5 polypeptide, was localized within the cochlea in situ to the cell types that form the lateral wall of the cochlear duct-the external sulcus (ES) cells and the cells of the spiral prominence. Expression of AQP5 was observed in the apical turn but not the basal turn of the cochlea; nor was it observed in the vestibular neuroepithelia or its supporting cells. The restricted expression of AQP5 to the apical turns of the cochlea suggests its potential role in low frequency hearing.

Animals↗

PubMed1996

Different effects of noise and salicylate and their interactions on the guinea pig cochlea.

We investigated the effects of noise and salicylate on the guinea pig cochlea by monitoring the acoustic nerve compound action potential (CAP) as well as the 2f1-f2 distortion product in the cochlear microphonics (DP-CM). The alterations of DP-CMs by noise and salicylate revealed an apparent difference, even when the noise-induced and salicylate-induced CAP threshold shifts were equal. This result indicated that noise and salicylate might have an effect on different sites in the cochlea. Combined effects were altered by changing the order in which the two agents were applied. The existing noise-induced damage attenuated the additional ototoxicity of salicylate. In contrast, attenuation did not occur when salicylate administration preceded noise exposure. Considering known effects of noise and/or salicylate altering mechano-electrical transduction (MET) and electro-mechanical transduction of the cochlear outer hair cells, our results seem to suggest that the MET plays a key role in the transduction mechanism in the cochlea.

Animals↗

PubMed1990

Regional variations in the expression of cytokeratin proteins in the adult human cochlea.

In the adult human cochlea, a cytokeratin (Ck) network exists along the entire surface of the organ of Corti, enclosing it like a shell. Only the surfaces of the outer and inner hair cells are not integrated in this network. In temporal bone specimens, Ck filaments in Hensen's cells were found to be arranged parallel with and closely apposed to the plasma membrane. In the stria vascularis, Cks were identified only in the marginal cells. Cells in Reissner's membrane and spiral prominence showed varying degrees of immunoreactivity to different monoclonal antibodies directed against Cks. A distinct positivity for Cks was found in most spiral ganglion cells, indicating their presence in all cells. The principal pattern of immunoreactivity was the same in the organ of Corti of the entire cochlea. However, a quantitative gradient in the expression of Cks was observed, with more Cks at the apex than at the base. This was correlated to a difference in the number of Hensen's cells between the two regions. The distinct shell configuration of the Ck network in Corti's organ gives it a tonotopically related difference in rigidity which must be of considerable importance for the perception of sound in the cochlea. The absence of Cks in inner and outer sulcus cells gives them cytoskeletal characteristics of mesenchymal cells with a possible regenerative potential.

Adult↗

PubMed1996

Combined effects of acute lead acetate exposure and tone exposure of the guinea pig cochlea.

Lead acetate exposure to humans can induce various disorders of the cranial nerves. Although vertigo and sensorineural deafness have been reported in lead workers, the dose effects of lead acetate on the cochlea and eighth cranial nerve are not well documented. We investigated the effects of lead acetate on the male albino Hartley guinea pig cochlea by measuring cochlear microphonics (CM), whole nerve action potential (AP), endocochlear potential (EP) and K+ ion concentration of the endolymph. Animals were given lead acetate by intraperitoneal injection as 20 mg/week for 4 consecutive weeks. A total dose < 80 mg did not induce electrophysiological changes in the cochlea. However, the AP output voltage (N1) decreased if the 80 mg lead acetate treatment was followed by an 80 dB tone exposure at 6 kHz during 24 h. A change was observed in CM and EP but not K+ ion concentration in the scala media.

Action Potentials↗

PubMed1994

Holmium: YAG laser surgery in obliterated cochleas: an experimental study in human cadaver temporal bones.

A pulsed holmium:YAG laser (lambda = 2120 nm) was used to reopen the basal turn of artificially obliterated human cochleas in freshly dissected cadavers. This allowed intracochlear insertion of the stimulation electrode of a cochlear implant under simulated surgical conditions. Laser energy was transmitted through a 400-microns nylon fiber via the opened facial recess directly to the round window niche. At an energy level of 500 mJ per 2.5 microseconds pulse, a repetition rate of 2 Hz, and an exposure time of 20-30 s, the photo-ablative mechanism of laser-bone interaction led to a 8-10 mm recanalization of the basal turn of the cochlea without damaging surrounding structures. The artificial bony occlusion and the recanalized basal turn of the cochlea were visualized by means of computed tomography and studied under light microscopy, using a histologic thin-section technique.

Aluminum Silicates↗

PubMed1992

Structure and function of the cochlea in the African mole rat (Cryptomys hottentotus): evidence for a low frequency acoustic fovea.

The cochlea of the mole rat Cryptomys hottentotus was investigated with physiological and anatomical methods. In order to reveal the place-frequency map of the cochlea, iontophoretic HRP-applications were made in the cochlear nucleus at physiologically characterized locations. Subsequent HRP-transport in auditory nerve fibres and labeling patterns of spiral ganglion cells within the cochlea were evaluated. A cochlear place-frequency map was constructed from 17 HRP-applications in the cochlear nucleus at positions where neurons had characteristic frequencies between 0.1 and 12.6 kHz. As in other mammals, high frequencies were found to be represented at the cochlear base, low frequencies at the cochlear apex. The place-frequency map had three distinct parts which were characterized by their different slopes. A clear over-representation of the frequencies between 0.6 and 1 kHz was revealed, in this frequency range the slope of the place-frequency map amounted to 5.3 mm/octave. As calculated from the regression analysis, below 0.6 kHz the slope of the cochlear place-frequency map amounted to 0.24 mm/octave, above 1 kHz to 0.9 mm/octave. As in other mammals width of the basilar membrane (BM) increased from the cochlear base towards the cochlear apex. Also in concordance with the findings in other mammals, BM-thickness decreased from the cochlear base to the apex. However, it was remarkable to find that there was no or little change in BM-width and thickness between 40 and 85% BM-length. It was also revealed that scala tympani was only 1/10th the size found in the rat or other mammals of similar body size.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1993

Late developmental changes of the innervation densities of the myelinated fibres and the outer hair cell efferent fibres in the rat cochlea.

The baso-apical distributions of the myelinated nerve fibres (representative for the inner hair cell afferent fibres) and the outer hair cell efferent fibres were studied during postnatal development of the rat cochlea. The myelinated fibres were counted in the primary osseos spiral lamina from semi-thin sections. The outer hair cell efferent fibres were counted in the tunnel of Corti by means of ultra-thin sections. The developmental changes of the myelinated fibres were investigated between 8 and 60 days after birth (DAB); those of the outer hair cell efferent fibres between 20 and 30 DAB. Between 12 DAB (onset of hearing) and 20 DAB the baso-apical distribution of the myelinated fibres does not change. Striking maturational changes occur later after the onset of hearing, between 20 and 30 DAB. The innervation density of the myelinated fibres increases in the lower middle region of the cochlea. In this region a maximum of innervation density appears. The efferent fibres to the outer hair cells show at 20 DAB a maximum of innervation density in the middle of the cochlea but between 20 and 30 DAB, the fibre density decreases in this region. During the same period the maximum of innervation density shifts towards the base. The change in the innervation densities of the myelinated fibres and the outer hair cell efferent fibres occurs late in development, after the onset of hearing, and after the organ of Corti shows an adult-like appearance.

Animals↗