The mechanism underlying maintenance of the endocochlear potential by the K+ transport system in fibrocytes of the inner ear.

The endocochlear potential (EP) of +80 mV in the scala media, which is indispensable for audition, is controlled by K+ transport across the lateral cochlear wall. This wall includes two epithelial barriers, the syncytium and the marginal cells. The former contains multiple cell types, such as fibrocytes, which are exposed to perilymph on their basolateral surfaces. The apical surfaces of the marginal cells face endolymph. Between the two barriers lies the intrastrial space (IS), an extracellular space with a low K+ concentration ([K+]) and a potential similar to the EP. This intrastrial potential (ISP) dominates the EP and represents the sum of the diffusion potential elicited by a large K+ gradient across the apical surface of the syncytium and the syncytium's potential, which is slightly positive relative to perilymph. Although a K+ transport system in fibrocytes seems to contribute to the EP, the mechanism remains uncertain. We examined the electrochemical properties of the lateral wall of guinea pigs with electrodes sensitive to potential and K+ while perfusing into the perilymph of the scala tympani blockers of Na+,K+-ATPase, the K+ pump thought to be essential to the system. Inhibiting Na+,K+-ATPase barely affected [K+] in the IS but greatly decreased [K+] within the syncytium, reducing the K+ gradient across its apical surface. The treatment hyperpolarized the syncytium only moderately. Consequently, both the ISP and the EP declined. Fibrocytes evidently use the Na+,K+-ATPase to achieve local K+ transport, maintaining the syncytium's high [K+] that is crucial for the K+ diffusion underlying the positive ISP.

Visualization of spiral ganglion neurites within the scala tympani with a cochlear implant in situ.

Current cochlear histology methods do not allow in situ processing of cochlear implants. The metal components of the implant preclude standard embedding and mid-modiolar sectioning, and whole mounts do not have the spatial resolution needed to view the implant within the scala tympani. One focus of recent auditory research is the regeneration of structures within the cochlea, particularly the ganglion cells and their processes, and there are multiple potential benefits to cochlear implant users from this work. To facilitate experimental investigations of auditory nerve regeneration performed in conjunction with cochlear implantation, it is critical to visualize the cochlear tissue and the implant together to determine if the nerve has made contact with the implant. This paper presents a novel histological technique that enables simultaneous visualization of the in situ cochlear implant and neurofilament-labeled nerve processes within the scala tympani, and the spatial relationship between them.

Stem cell transplantation for auditory nerve replacement.

The successful function of cochlear prostheses depends on activation of auditory nerve. The survival of auditory nerve neurons, however, can vary widely in candidates for cochlear implants and influence implant efficacy. Stem cells offer the potential for improving the function of cochlear prostheses and increasing the candidate pool by replacing lost auditory nerve. The first phase of studies for stem cell replacement of auditory nerve has examined the in vitro survival and differentiation as well as in vivo differentiation and survival of exogenous embryonic and tissue stem cells placed into scala tympani and/or modiolus. These studies are reviewed and new results on in vivo placement of B-5 mouse embryonic stem cells into scala tympani of the guinea pig cochleae with differentiation into a glutamatergic neuronal phenotype are presented. Research on the integration and connections of stem cell derived neurons in the cochlea is described. Finally, an alternative approach is considered, based on the use of endogenous progenitors rather than exogenous stem cells, with a review of promising findings that have identified stem cell-like progenitors in cochlear and vestibular tissues to provide the potential for auditory nerve replacement.

Survival of neuronal tissue following xenograft implantation into the adult rat inner ear.

The poor regenerative capacity of the spiral ganglion neurons of the mammalian inner ear has initiated research on how to assist the functional recovery of the injured auditory system. A possible treatment is to use a biological implant with a potential to establish central or peripheral synaptic contacts to develop into a functional auditory unit. The feasibility of this approach was tested by xenograft implantation of dorsal root ganglion (DRG) neurons from embryonic days 13 to 14, mouse expressing either LacZ or enhanced green fluorescent protein (EGFP) into the scala tympani of the adult rat inner ear. Transplanted DRG neurons survived in the scala tympani for a postoperative period ranging from 3 to 10 weeks, as verified by histochemical detection of LacZ, EGFP fluorescence and immunohistochemical labeling of the neuronal markers neurofilament and Thy 1.2. DRG neurons were found close to structures near the sensory epithelium (the organ of Corti) as well as adjacent to the spiral ganglion neurons with their peripheral dendrites. These results illustrate not only the survival of xenografted DRG neurons in the adult inner ear but also the feasibility of a neuronal transplantation strategy in the degenerated auditory system, thereby creating possibilities to replace spiral ganglion neurons.

Internal shearing within the hearing organ evoked by basilar membrane motion.

The vibration of the hearing organ that occurs during sound stimulation is based on mechanical interactions between different cellular structures inside the organ of Corti. The exact nature of these interactions is unclear and subject to debate. In this study, dynamic structural changes were produced by stepwise alterations of scala tympani pressure in an in vitro preparation of the guinea pig temporal bone. Confocal images were acquired at each level of pressure. In this way, the motion of several structures could be observed simultaneously with high resolution in a nearly intact system. Images were analyzed using a novel wavelet-based optical flow estimation algorithm. Under these conditions, the reticular lamina moved as a stiff plate with a center of rotation in the region of the inner hair cells. Despite being enclosed in several types of supporting cells, the inner hair cells, together with the adjacent inner pillar cells, moved in a manner signifying high compliance. The outer hair cells displayed radial motion indicative of cellular bending. Together, these results show that shearing motion occurs between several parts of the organ, and that structural relationships within the organ change dynamically during displacement of the basilar membrane.

Induction of macrophage exudation in the inner ear by OK432 treatment.

OK432, a heat- and penicillin-treated lyophilized powder of a low virulent Su strain of Streptococcus pyogenes, was injected into the perilymphatic space of guinea pigs in order to determine the kinetics of exudation of inflammatory cells into the cochlea over a 7-day period. OK432 induced exudation of many neutrophils and Asialo GM1-positive cells into the scala tympani. The numbers of these cells peaked on the first day after OK432 treatment, and then gradually decreased. Four days after treatment very few of either of these types of cells were observed in the scala tympani. Asialo GM1-positive cells were confirmed to be activated macrophages. In ears treated with physiological saline or with the additive included in OK432 in control studies, no notable changes in the inner ear were recognized. These findings suggest that the inner ear can undergo induction of macrophage rapidly exudation by OK432 treatment as has previously been observed for other organs.

Asialo GM1-positive cells in mouse cochlea.

The presence of ganglio-N-tetraosylceramide (asialo GM1)-positive cells in mouse cochlea was tested for using an immunohistochemical method with an anti-asialo GM1 antibody which reacts with natural killer cells and macrophages in mouse. Only a few asialo GM1-positive cells were present on the surface of the scala tympani of the basal turn near the collecting venules. Transmission electron microscopic study showed the internal structure of these cells to be that of macrophages. These findings suggest that asialo GM1-positive cells function as resident type macrophages against invasion by pathogens of the mouse cochlea.

Bone lining cells of the mammalian cochlea.

It is generally believed that all bone surfaces are covered by a nearly continuous layer of cells known as bone lining cells (BLC) which separate the general extracellular fluid (GECF) from bone and its fluid compartment. Within the cochlea of some mammals regions of bone matrix are exposed to extracellular fluid. Within the scalae of the cochlea, perilymph is in contact with bone matrix; there is no evidence of a lining endothelium. Within the modiolus and subjacent to the spiral ligament, bone matrix is in contact with GECF. These findings may have importance in understanding calcium homeostasis within the scalae and may relate to the pathophysiology of labyrinthitis ossificans. Additionally, since BLCs probably represent a specific phenotype, the presence of a pure population of BLCs within the scalae may provide a source for the development of a pure culture of this cell.

The occurrence of capillary endothelial mitoses in the mesenchyme of the scala tympani during fetal development of the guinea pig.

Mitotic figures were found in the capillary endothelial cells of the cochleae of guinea pig fetuses, indicating a cellular proliferation. The significance of capillary sprouting in the mesenchyme beneath the basilar membrane a few days before opening of the scala tympani is discussed.