The microenvironment of the cochlea is maintained by the barrier between the systemic circulation and the fluids inside the stria vascularis. However, the mechanisms that control the permeability of the intrastrial fluid-blood barrier remain largely unknown. The barrier comprises endothelial cells connected to each other by tight junctions and an underlying basement membrane. In a recent study, we found that the intrastrial fluid-blood barrier also includes a large number of perivascular cells with both macrophage and melanocyte characteristics. The perivascular-resident macrophage-like melanocytes (PVM/Ms) are in close contact with vessels through cytoplasmic processes. Here we demonstrate that PVM/Ms have an important role in maintaining the integrity of the intrastrial fluid-blood barrier and hearing function. Using a cell culture-based in vitro model and a genetically induced PVM/M-depleted animal model, we show that absence of PVM/Ms increases the permeability of the intrastrial fluid-blood barrier to both lowand high-molecular-weight tracers. The increased permeability is caused by decreased expression of pigment epithelial-derived factor, which regulates expression of several tight junction-associated proteins instrumental to barrier integrity. When tested for endocochlear potential and auditory brainstem response, PVM/ M-depleted animals show substantial drop in endocochlear potential with accompanying hearing loss. Our results demonstrate a critical role for PVM/Ms in regulating the permeability of the intrastrial fluid-blood barrier for establishing a normal endocochlear potential hearing threshold. mouse cochlea | paracellular permeability | tight junction | capillary T he intrastrial fluid-blood barrier separates the stria vascularis (SV) from peripheral circulation. The integrity of the barrier is critical for maintaining inner ear homeostasis, especially for sustaining the endocochlear potential (EP), an essential driving force for hearing function (1-4). Disruption of the barrier is closely associated with a number of hearing disorders, including autoimmune inner ear disease, noise-induced hearing loss, agerelated hearing loss, and several genetically linked diseases (5-10). Despite the importance of the intrastrial fluid-blood barrier, little is understood about regulation of the barrier and the mechanisms that control its permeability.In the classic view, the intrastrial fluid-blood barrier comprises basement membrane and endothelial cells (ECs) that connect to each other with tight junctions (11) to form a diffusion barrier that prevents most blood-borne substances from entering the ear (2). In a recent study, we found that the intrastrial fluid-blood barrier also includes a large number of pericytes and perivascular-resident macrophage-like melanocytes (PVM/Ms) (12, 13). The PVM/Ms are not observed in other capillary regions such as in capillary beds of the spiral ligament. The PVM/Ms are in close contact with vessels through cytoplasmic processes. The structural complexity of PVM/Ms' capillar...

The ear is a remarkably sensitive pressure fluctuation detector. In guinea pigs, behavioral measurements indicate a minimum detectable sound pressure of ~20 μPa at 16 kHz. Such faint sounds produce 0.1 nm basilar membrane displacements, a distance smaller than conformational transitions in ion channels. It seems that noise within the auditory system would swamp such tiny motions, making weak sounds imperceptible. Here, a new mechanism contributing to a resolution of this problem is proposed and validated through direct measurement. We hypothesize that vibration at the apical end of hair cells is enhanced compared to the commonly measured basilar membrane side. Using in vivo optical coherence tomography, we demonstrated that apical-side vibrations peak at a higher frequency, had different timing, and were enhanced compared to the basilar membrane. These effects depend nonlinearly on the stimulus level. The timing difference and enhancement are important for explaining how the noise problem is circumvented.

The dynamic responses of the hearing organ to acoustic overstimulation were investigated using the guinea pig isolated temporal bone preparation. The organ was loaded with the f luorescent Ca 2؉ indicator Fluo-3, and the cochlear electric responses to low-level tones were recorded through a microelectrode in the scala media. After overstimulation, the amplitude of the cochlear potentials decreased significantly. In some cases, rapid recovery was seen with the potentials returning to their initial amplitude. ] changes were not seen in preparations that were stimulated at levels that did not cause an amplitude change in the cochlear potentials. The overstimulation also gave rise to a contraction, evident as a decrease of the width of the organ of Corti. The average contraction in 10 preparations was 9 m (SE 2 m). Partial or complete recovery was seen within 30-45 min after the overstimulation. The [Ca 2؉ ] changes and the contraction are likely to produce major functional alterations and consequently are suggested to be a factor contributing strongly to the loss of function seen after exposure to loud sounds.Noise-induced hearing loss is a common condition that leads to considerable communication problems for affected individuals. Recent research on the physiology of this condition (reviewed in ref. 1) has been mainly focused on damage to the stereocilia (SC) of the sensory cells in the inner ear, important because this is the location of the ion channels converting mechanic vibrations into electric currents. Damage to the SC correlates well with alterations of the tuning curves of auditory nerve fibres (2). A capacity for repair of the SC after acoustic trauma also has been implicated (3), but the mechanisms underlying the stereociliary changes as well as the repair process remain unknown.Acoustic trauma also may cause degeneration of the sensory cells, resulting in an irreversible elevation in hearing thresholds (4). The degeneration most likely involves not only stereociliary changes but also alterations at the cell body level. The events taking place in these cells during and after overstimulation remain largely obscure. Cody and Russell (5) have shown that sustained depolarizations of the outer hair cells (OHCs) occur after moderately intense acoustic overstimulation and that repolarization parallels the recovery of auditory sensitivity. The underlying mechanisms are unclear.In isolated OHCs, mechanical overstimulation results in cytoplasmic [Ca 2ϩ ] increase (6). To investigate how this finding relates to reduced hearing sensitivity after acoustic trauma, the guinea pig isolated temporal bone preparation (7) was used to perform simultaneous investigations of calcium-dependent fluorescence, stimulus-evoked cochlear potentials and cochlear morphology. The sensory cells were visualized in situ in an almost native environment, and the cochlear electric responses were recorded. The preparation was used previously to study changes of organ of Corti mechanics following acoustic trauma (8) and has now been ...

During sound stimulation, receptor potentials are generated within the sensory hair cells of the cochlea. Prevailing theory states that outer hair cells use the potential-sensitive motor protein prestin to convert receptor potentials into fast alterations of cellular length or stiffness that boost hearing sensitivity almost 1000-fold. However, receptor potentials are attenuated by the filter formed by the capacitance and resistance of the membrane of the cell. This attenuation would limit cellular motility at high stimulus frequencies, rendering the above scheme ineffective. Therefore, Dallos and Evans (1995a) proposed that extracellular potential changes within the organ of Corti could drive cellular motor proteins. These extracellular potentials are not filtered by the membrane. To test this theory, both electric potentials inside the organ of Corti and basilar membrane vibration were measured in response to acoustic stimulation. Vibrations were measured at sites very close to those interrogated by the recording electrode using laser interferometry. Close comparison of the measured electrical and mechanical tuning curves and time waveforms and their phase relationships revealed that those extracellular potentials indeed could drive outer hair cell motors. However, to achieve the sharp frequency tuning that characterizes the basilar membrane, additional mechanical processing must occur inside the organ of Corti.