Downregulation of Cav3.1 T-type Calcium Channel Expression in Age-related Hearing Loss Model.

OBJECTIVE: Age-related hearing loss (AHL), characterized by degeneration of cochlea structures, is the most common sensory disorder among the elderly worldwide. The calcium channel is considered to contribute to normal hearing. However, the role of the T-type voltage-activated calcium channel, Cav3.1, remains unclear in AHL. Here, we investigate the age-related change of Cav3.1 expression in the cochlea and D-gal-induced senescent HEI-OC1 cells. METHODS: Cochleae from C57BL/6 mice at 2 months and 12 months of age were assessed. Senescence in House Ear Institute-Organ of Corti 1 (HEI-OC1) cells was induced by D-gal treatment. The immunofluorescence technique was employed to investigate the distribution of Cav3.1 in vivo and in vitro. Quantitative assessment was achieved by Western blotting and real-time PCR. RESULTS: In comparison with 2-month-old animals, 12-month old C57BL/6 mice exhibited great loss of hair cells and elevated auditory brainstem threshold. The Cav3.1 was located in hair cells, spiral ganglion cells, lateral walls, and the expression of Cav3.1 protein and mRNA decreased in the aged cochleae. D-gal-induced senescence assay confirmed the down-regulation of Cav3.1 expression in senescent HEI-OC1 cells. CONCLUSION: Our results show that age-related down-regulated expression of Cav3.1 in the cochleae is associated with AHL and may contribute to the pathogenesis of AHL.

Cellular cartography of the organ of Corti based on optical tissue clearing and machine learning.

The highly organized spatial arrangement of sensory hair cells in the organ of Corti is essential for inner ear function. Here, we report a new analytical pipeline, based on optical clearing of tissue, for the construction of a single-cell resolution map of the organ of Corti. A sorbitol-based optical clearing method enabled imaging of the entire cochlea at subcellular resolution. High-fidelity detection and analysis of all hair cell positions along the entire longitudinal axis of the organ of Corti were performed automatically by machine learning-based pattern recognition. Application of this method to samples from young, adult, and noise-exposed mice extracted essential information regarding cellular pathology, including longitudinal and radial spatial characteristics of cell loss, implying that multiple mechanisms underlie clustered cell loss. Our method of cellular mapping is effective for system-level phenotyping of the organ of Corti under both physiological and pathological conditions.

Organ of Corti vibration within the intact gerbil cochlea measured by volumetric optical coherence tomography and vibrometry.

There is indirect evidence that the mammalian cochlea in the low-frequency apical and the more commonly studied high-frequency basal regions function in fundamentally different ways. Here, we directly tested this hypothesis by measuring sound-induced vibrations of the organ of Corti (OoC) at three turns of the gerbil cochlea using volumetric optical coherence tomography vibrometry (VOCTV), an approach that permits noninvasive imaging through the bone. In the apical turn, there was little frequency selectivity, and the displacement-vs.-frequency curves had low-pass filter characteristics with a corner frequency of ~0.5-0.9 kHz. The vibratory magnitudes increased compressively with increasing stimulus intensity at all frequencies. In the middle turn, responses were similar except for a slight peak in the response at ~2.5 kHz. The gain was ~50 dB at the peak and 30-40 dB at lower frequencies. In the basal turn, responses were sharply tuned and compressively nonlinear, consistent with observations in the literature. These data demonstrated that there is a transition of the mechanical response of the OoC along the length of the cochlea such that frequency tuning is sharper in the base than in the apex. Because the responses are fundamentally different, it is not appropriate to simply frequency shift vibratory data measured at one cochlear location to predict the cochlear responses at other locations. Furthermore, this means that the number of hair cells stimulated by sound is larger for low-frequency stimuli and smaller for high-frequency stimuli for the same intensity level. Thus the mechanisms of central processing of sounds must vary with frequency. NEW & NOTEWORTHY A volumetric optical coherence tomography and vibrometry system was used to probe cochlear mechanics within the intact gerbil cochlea. We found a gradual transition of the mechanical response of the organ of Corti along the length of the cochlea such that tuning at the base is dramatically sharper than that at the apex. These data help to explain discrepancies in the literature regarding how the cochlea processes low-frequency sounds.

Evaluation of Cochlear Duct Length Computations Using Synchrotron Radiation Phase-Contrast Imaging.

HYPOTHESIS: Evaluation of cochlear duct length (CDL) using novel imaging techniques will help improve the accuracy of existing CDL equations. BACKGROUND: Various relationships relating A value measured from a patient's computed tomography scan and CDL have been proposed to aid in preoperative electrode selection and frequency mapping. METHODS: Ten cadaveric temporal bones were scanned using synchrotron radiation phase-contrast imaging. Reference CDL values were calculated by placing points representing the organ of Corti (OC), lateral wall (LW), and electrode location (I) on the synchrotron radiation phase-contrast imaging slices along the length of the cochlea. The CDL estimates from the existing three equations (OC, LW, I) in addition to two newly proposed equations (OC and LW) were compared with reference CDL values at each respective location. RESULTS: When compared with reference CDL values, the new OC equation improved the CDL estimates from a 6.2% error to a 5.1% error while the new LW equation improved the CDL estimate error from 3.9 to 3.6%. Bland-Altman plots revealed both new equations increased similarity to reference values and brought more samples to within clinically significant ranges. Validation of the original electrode location equation to the reference values showed a 4.6% difference. CONCLUSION: The newly proposed equations for LW and OC provided an improvement over past equations for determining CDL from the A value by showing improved agreement with reference values. Therefore, these equations can provide quick and accurate preoperative estimates of CDL for improving customized frequency mapping.

Micro-optical coherence tomography of the mammalian cochlea.

The mammalian cochlea has historically resisted attempts at high-resolution, non-invasive imaging due to its small size, complex three-dimensional structure, and embedded location within the temporal bone. As a result, little is known about the relationship between an individual's cochlear pathology and hearing function, and otologists must rely on physiological testing and imaging methods that offer limited resolution to obtain information about the inner ear prior to performing surgery. Micro-optical coherence tomography (µOCT) is a non-invasive, low-coherence interferometric imaging technique capable of resolving cellular-level anatomic structures. To determine whether µOCT is capable of resolving mammalian intracochlear anatomy, fixed guinea pig inner ears were imaged as whole temporal bones with cochlea in situ. Anatomical structures such as the tunnel of Corti, space of Nuel, modiolus, scalae, and cell groupings were visualized, in addition to individual cell types such as neuronal fibers, hair cells, and supporting cells. Visualization of these structures, via volumetrically-reconstructed image stacks and endoscopic perspective videos, represents an improvement over previous efforts using conventional OCT. These are the first µOCT images of mammalian cochlear anatomy, and they demonstrate µOCT's potential utility as an imaging tool in otology research.

Kanamycin-furosemide ototoxicity in the mouse cochlea: a 3-dimensional analysis.

OBJECTIVE: Administration of an aminoglycoside antibiotic and loop diuretic causes damage to hair cells in the organ of Corti, resulting in their death and the death of their corresponding spiral ganglion neurons. While this phenomenon has been studied previously, analysis of its effects in the whole cochlea has not been reported. The authors sought to evaluate the effects of a combination dose of kanamycin and furosemide in mice cochlea using an imaging system and computer analysis that allowed for nondestructive, whole-cochlea visualization. STUDY DESIGN: Study using an animal model. SETTING: Cochlear analysis laboratory. SUBJECTS AND METHODS: Five mice received kanamycin and furosemide and 3 mice received saline. Cochleas were harvested and imaged with scanning thin-sheet laser imaging microscopy (sTSLIM) to analyze sensory cells and cochlea structures. RESULTS: The drug-treated animals showed substantial loss of inner hair cells and complete outer hair cell loss. All treated mice showed spiral ganglion neuron loss with fewer neurons than control animals and decreased cell density in the middle turn of the cochlea. The spiral ligament and spiral limbus in the treated animals also showed a decrease in fibrocyte cell density in the middle to apical portion of the cochlea. The stria vascularis appeared normal in all animals. CONCLUSION: Imaging methods that allow for whole-cochlea analysis provide insight into changes that occur in the cochlea after ototoxic insult. Trends that may not be apparent in cross-section samples of the cochlea can be observed. Computer analysis of these trends allows them to be assessed accurately.

Cochlear coordinates in regard to cochlear implantation: a clinically individually applicable 3 dimensional CT-based method.

SETTING: Cochlear implant (CI)/tertiary referral center. SUBJECTS: Twenty-five patients implanted with an Advanced Bionics HiRes90K HiFocus1J CI. STUDY DESIGN/MAIN OUTCOME MEASURES: A 3-dimensional cylindrical coordinate system is introduced using the basal turn of the cochlea as the x and y planes and the center of the modiolus as the z axis. The 0-degree angle is defined by the most lateral point of the horizontal semicircular canal. It is applied to both preoperative and postoperative computed tomographies in 25 patients. The angular position of the round window is examined. Interobserver reproducibility is tested by localization of all electrode contacts within the coordinate system. To observe realignment over time, electrode coordinates in postoperative images were projected on preoperative images. Additionally, comparison to existing imaging-related coordinate systems was made. RESULTS: The angular position of the center of the round window is 34.6 +/- 0.4 degrees (standard deviation) with an intraclass coefficient of 1.00. The intraclass coefficient for interobserver reproducibility of the 16 electrode contacts ranged from 0.74 to 1 for the rotational angle (phi) and 0.77 to 1 for the distance to the modiolus (rho). In 21 of 25 patients, a perfect match or minimal displacement of up to 3 electrode contacts was seen. Comparison to existing systems showed good correlation. CONCLUSION: A 3-dimensional cochlear coordinate system easily applicable in clinical patients is described, which fulfills the requirements set by an international consensus.

Imágenes en anatomía coclear TC y RM / Cochlear anatomy CT and MR imaging

Los autores ofrecen un breve repaso de anatomía normal de la cóclea con imágenes de TC y RM a fin de posibilitar una más acabada identificación de hallazgos patológicos en pacientes con hipoacusia perceptiva (AU)

Furosemide alters organ of corti mechanics: evidence for feedback of outer hair cells upon the basilar membrane.

A widely held hypothesis of mammalian cochlear function is that the mechanical responses to sound of the basilar membrane depend on transduction by the outer hair cells. We have tested this hypothesis by studying the effect upon basilar membrane vibrations (measured by means of either the Mössbauer technique or Doppler-shift laser velocimetry) of systemic injection of furosemide, a loop diuretic that decreases transduction currents in hair cells. Furosemide reversibly altered the responses to tones and clicks of the chinchilla basilar membrane, causing response-magnitude reductions that were largest (up to 61 dB, averaging 25-30 dB) at low stimulus intensities at the characteristic frequency (CF) and small or nonexistent at high intensities and at frequencies far removed from CF. Furosemide also induced response-phase lags that were largest at low stimulus intensities (averaging 77 degrees) and were confined to frequencies close to CF. These results constitute the most definitive demonstration to date that mechanical responses of the basilar membrane are dependent on the normal function of the organ of Corti and strongly implicate the outer hair cells as being responsible for the high sensitivity and frequency selectivity of basilar membrane responses. A corollary of these findings is that sensorineural hearing deficits in humans due to outer hair cell loss reflect pathologically diminished vibrations of the basilar membrane.