Vestibular Endolymph Research Articles

Magnetic Resonance Volumetric Quantification of Vestibular Endolymphatic Hydrops in Patients with Unilateral Definite Meniere's Disease Using 3D Inversion Recovery with Real Reconstruction (3D-REAL-IR) Sequence.

The 3D-REAL-IR MRI sequence allows for an in vivo visualization of endolymphatic hydrops. Qualitative assessment methods of the severity of vestibular and cochlear hydrops are the most commonly used. A quantitative volumetric measurement of vestibular EH in patients with definite unilateral Ménière's disease using the 3D-REAL-IR sequence and the calculation of the endolymphatic ratio (ELR) was intended. Volumetric calculations of the vestibules, vestibular endolymph and vestibular ELR are performed in 96 patients with unilateral Ménière's disease and correlated with classic qualitative grading scales. Quantitative volumetric measurement of vestibular hydrops using the 3D-REAL-IR sequence is feasible and reproducible in daily clinical practice. Vestibular ELR values exceeding 60% defined radiologically significant vestibular hydrops, while values below 30% defined radiologically non-significant vestibular hydrops.

Volume quantification of endolymphatic hydrops in patients with vestibular schwannoma

ObjectiveThe origin of vestibular symptoms in patients with vestibular schwannoma (VS) is uncertain. We used intratympanic gadolinium-enhanced magnetic resonance imaging (MRI) to confirm the labyrinthine lesions in patients with VS and to explore the features of endolymphatic hydrops (EH) in these patients. MethodsIn total, 66 patients diagnosed with unilateral VS were enrolled in this study and underwent intratympanic gadolinium-enhanced MRI. The borders of the vestibule and endolymph were mapped on the axial MRI images, and the area and volume of vestibule and endolymph were automatically calculated using Osirix software, and the area and volume percentage of vestibular endolymph were obtained. ResultsThe area and volume percentages of vestibular endolymph on the affected side were significantly larger than those on the healthy side (both p < 0.001). Using Kendall's W test, we found that the area and volume percentages of vestibular endolymph on the affected side were consistent (p < 0.001), but the consistency was moderate (k = 0.574). The healthy side was also consistent (p < 0.001), and the degree of consistency was moderate (k = 0.444). We used 19.1% as the cut-off point to distinguish the presence or absence of vestibular EH; that is, the volume percentage of vestibular endolymph that was more than 19.1% were defined as the subgroup with hydrops, while the subgroup without hydrops included patients with a baseline level below 19.1%. No volume classification for vestibular EH was proposed. Based on this standard, 11/66 (16.7%) of the patients with VS in this study had vestibular EH. Conclusions.The volume percentage of the vestibular endolymph was more accurate than the area percentage for assessing vestibular EH. Using 19.1% as the cut-off point to distinguish the presence or absence of vestibular EH, we found that 16.7% of patients with VS had varying degrees of vestibular EH. We believe that the vestibular symptoms in patients with VS may originate from the peripheral lesions.

Endolymphatic Potential Measured From Developing and Adult Mouse Inner Ear.

The mammalian inner ear has two major parts, the cochlea is responsible for hearing and the vestibular organ is responsible for balance. The cochlea and vestibular organs are connected by a series of canals in the temporal bone and two distinct extracellular fluids, endolymph and perilymph, fill different compartments of the inner ear. Stereocilia of mechanosensitive hair cells in the cochlea and vestibular end organs are bathed in the endolymph, which contains high K+ ions and possesses a positive potential termed endolymphatic potential (ELP). Compartmentalization of the fluids provides an electrochemical gradient for hair cell mechanotransduction. In this study, we measured ELP from adult and neonatal C57BL/6J mice to determine how ELP varies and develops in the cochlear and vestibular endolymph. We measured ELP and vestibular microphonic response from saccules of neonatal mice to determine when vestibular function is mature. We show that ELP varies considerably in the cochlear and vestibular endolymph of adult mice, ranging from +95 mV in the basal turn to +87 mV in the apical turn of the cochlea, +9 mV in the saccule and utricle, and +3 mV in the semicircular canal. This suggests that ELP is indeed a local potential, despite the fact that endolymph composition is similar. We further show that vestibular ELP reaches adult-like magnitude around post-natal day 6, ~12 days earlier than maturation of cochlear ELP (i.e., endocochlear potential). Maturation of vestibular ELP coincides with the maturation of vestibular microphonic response recorded from the saccular macula, suggesting that maturation of vestibular function occurs much earlier than maturation of hearing in mice.

Effect of head pitch and roll orientations on magnetically induced vertigo.

Key points Lying supine in a strong magnetic field, such as in magnetic resonance imaging scanners, can induce a perception of whole‐body rotation.The leading hypothesis to explain this invokes a Lorentz force mechanism acting on vestibular endolymph that acts to stimulate semicircular canals.The hypothesis predicts that the perception of whole‐body rotation will depend on head orientation in the field.Results showed that the direction and magnitude of apparent whole‐body rotation while stationary in a 7 T magnetic field is influenced by head orientation.The data are compatible with the Lorentz force hypothesis of magnetic vestibular stimulation and furthermore demonstrate the operation of a spatial transformation process from head‐referenced vestibular signals to Earth‐referenced body motion.

Ratio of Vestibular Endolymph in Patients with Isolated Lateral Semicircular Canal Dysplasia.

Isolated vestibular-lateral semicircular canal dysplasia (LSCCD) is one of the most common anomalies of the inner ear. However, endolymphatic size in LSCCD is unknown. We measured the size of the endolymph in the vestibule of patients with LSCCD and compared it with that measured in patients without LSCCD. We extracted 1102 magnetic resonance (MR) studies for the evaluation of endolymphatic hydrops (EH) from our database of radiology reports. Among these, we found 15 ears from 11 patients with LSCCD; 4 patients had bilateral abnormalities. Seven of the 15 ears demonstrated aplasia and 8 ears, hypoplasia of the lateral semicircular canal (LSCC). The control group consisted of 26 ears from 13 randomly selected patients without LSCCD. We measured the area of endolymph in the vestibule (ELA), total area of vestibular lymph fluid (TLA), and area of the central bony island (CBI) of the LSCC from axial MR images obtained after intratympanic or intravenous administration of gadolinium-based contrast material. The ratio of endolymphatic area to total lymphatic area (%EL) was defined as %EL = ELA/TLA × 100.We evaluated the correlation between %EL and the area of the CBI and compared age, %EL, degree of cochlear EH, hearing level, and presence of rotating vertigo among the 3 groups (aplasia, hypoplasia, control). The mean %EL was 76.7% in the aplasia group, 50.0% in the hypoplasia group, and 27.8% in the control group (P < 0.001). There was a relatively strong linear correlation between the area of the CBI and %EL (r = -0.767). Patient age, mean hearing level, degree of cochlear EH, or presence of vertigo attacks did not differ significantly among the groups (P > 0.05). The size of vestibular endolymph was larger in the groups with aplasia or hypoplasia than the control group. Thus, the current diagnostic cut-off value for significant vestibular EH (>50%) might not be appropriate for ears with LSCCD.

Bacterial Invasion of the Inner Ear in Association With Pneumococcal Meningitis

To examine the pathways of bacterial invasion and subsequent spreading in the inner ear during pneumococcal meningitis. A well-established adult rat model of Streptococcus pneumoniae meningitis was used. Thirty rats were inoculated intrathecally with S. pneumoniae serotype 1, 3 or 9 V and received no additional treatment. The rats were sacrificed when reaching terminal illness or on Day 7 and then prepared for serial sectioning and PAS-Alcian blue staining for light microscopy. During the first few days after inoculation, bacteria invade the inner ear through the cochlear aqueduct, into the scala tympani of the cochlea (perilymphatic space). From here, bacteria spreads apically toward the helicotrema and subsequently basally through the scala vestibuli, toward the vestibule and the vestibular system. When the bacteria after 5 to 6 days had reached scala vestibuli of the basal turn of the cochlea, hematogenous spreading occurred to the spiral ligament and into the cochlear endolymph, subsequently to the vestibular endolymph. We found no evidence of alternative routes for bacterial invasion in the inner ear. Several internal barriers to bacterial spreading were found within the inner ear. Bacterial elimination was evidenced by engulfment by macrophages within the inner ear. From the meninges, pneumococci invade the inner ear through the cochlear aqueduct during the first days of infection, whereas hematogenous invasion via the spiral ligament capillary bed occur at later stages. Although internal barriers exist within the inner ear, the spreading of bacteria occurs via the natural pathways of the fluid compartments. Bacterial elimination occurs by local macrophage engulfment.

In Vivo Visualized Endolymphatic Hydrops and Inner Ear Functions in Patients With Electrocochleographically Confirmed Ménière’s Disease

To study the correlation between the degree of endolymphatic hydrops as detected in vivo by magnetic resonance (MR) imaging and the auditory and vestibular function in patients with definite Ménière's disease. Prospective observational study. Tertiary referral center for balance disorders. In this prospective study, 41 patients who fulfilled the criteria for definite unilateral Ménière's disease according to the American Association of Otolaryngology-Head and Neck Surgery and who showed a summating potential-to-action potential ratio of greater than 0.4 on electrocochleography were included. Intratympanic contrast-enhanced 3 Tesla MR imaging of the inner ear was performed using a 3D Inversion Recovery Turbo Spin Echo sequence. The degree of endolymphatic hydrops was graded on a Likert scale (0-3) in the cochlea and by vestibular endolymph space ratio in the vestibulum. The degree of hydrops was then analyzed with respect to its correlation with audiometric hearing levels, interaural amplitude ratios of vestibular evoked myogenic potentials, degree of horizontal semicircular canal paresis on caloric irrigation, and disease duration. The degree of hearing loss and the disease duration correlated significantly with cochlear hydrops (r = 0.85; r = 0.34). Sacculus dysfunction was significantly correlated with vestibular hydrops (r = -0.47). There was no significant correlation between horizontal semicircular canal paresis and vestibular hydrops. In patients with clinically and electrocochleographically confirmed definite Ménière's disease, the degree of MR morphological hydrops severity correlates significantly with impairment of hearing function and sacculus function.

Imaging of Endolymphatic and Perilymphatic Fluid after Intravenous Administration of Single-dose Gadodiamide

To visualize endolymph as bright signal after intravenous injection of single-dose gadodiamide, we shortened the inversion time of heavily T(2)-weighted 3-dimensional (3D) fluid-attenuated inversion recovery (FLAIR) to 2050 ms. In 14 patients with suspected Ménière's disease, we observed high signal of vestibular endolymph in all ears, including 6 ears without vestibular endolymphatic hydrops. We observed high signal of cochlear endolymph in 17 ears with cochlear endolymphatic hydrops but not 11 ears without cochlear endolymphatic hydrops.

Critical Roles of Transitional Cells and Na/K-ATPase in the Formation of Vestibular Endolymph

The mechanotransduction of vestibular sensory cells depends on the high endolymphatic potassium concentration ([K+]) maintained by a fine balance between K+ secretion and absorption by epithelial cells. Despite the crucial role of endolymph as an electrochemical motor for mechanotransduction, little is known about the processes that govern endolymph formation. To address these, we took advantage of an organotypic rodent model, which regenerates a genuine neonatal vestibular endolymphatic compartment, facilitating the determination of endolymphatic [K+] and transepithelial potential (Vt) during endolymph formation. While mature Vt levels are almost immediately achieved, K+ accumulates to reach a steady [K+] by day 5 in culture. Inhibition of sensory cell K+ efflux enhances [K+] regardless of the blocker used (FM1.43, amikacin, gentamicin, or gadolinium). Targeting K+ secretion with bumetanide partially and transiently reduces [K+], while ouabain application and Kcne1 deletion almost abolishes it. Immunofluorescence studies demonstrate that dark cells do not express Na-K-2Cl cotransporter 1 (the target of bumetanide) in cultured and young mouse utricles, while Na/K-ATPase (the target of ouabain) is found in dark cells and transitional cells. This global analysis of the involvement of endolymphatic homeostasis actors in the immature organ (1) confirms that KCNE1 channels are necessary for K+ secretion, (2) highlights Na/K-ATPase as the key endolymphatic K+ provider and shows that Na-K-2Cl cotransporter 1 has a limited impact on K+ influx, and (3) demonstrates that transitional cells are involved in K+ secretion in the early endolymphatic compartment.

Sodium selectivity of semicircular canal duct epithelial cells

BackgroundSodium absorption by semicircular canal duct (SCCD) epithelial cells is thought to contribute to the homeostasis of the volume of vestibular endolymph. It was previously shown that the epithelial cells could absorb Na+ under control of a glucocorticoid hormone (dexamethasone) and the absorptive transepithelial current was blocked by amiloride. The most commonly-observed target of amiloride is the epithelial sodium channel (ENaC), comprised of the three subunits α-, β- and γ-ENaC. However, other cation channels have also been observed to be sensitive in a similar concentration range. The aim of this study was to determine whether SCCD epithelial cells absorb only Na+ or also K+ through an amiloride-sensitive pathway. Parasensory K+ absorption could contribute to regulation of the transduction current through hair cells, as found to occur via vestibular transitional cells [S. H. Kim and D. C. Marcus. Regulation of sodium transport in the inner ear. Hear.Res. doi:10.1016/j.heares.2011.05.003, 2011].ResultsWe determined the molecular and functional expression of candidate cation channels with gene array (GEO GSE6197), whole-cell patch clamp and transepithelial recordings in primary cultures of rat SCCD. α-, β- and γ-ENaC were all previously reported as present. The selectivity of the amiloride-sensitive transepithelial and cell membrane currents was observed in Ussing chamber and whole-cell patch clamp recordings. The cell membrane currents were carried by Na+ but not K+, but the Na+ selectivity disappeared when the cells were cultured on impermeable supports. Transepithelial currents across SCCD were also carried exclusively by Na+.ConclusionsThese results are consistent with the amiloride-sensitive absorptive flux of SCCD mediated by a highly Na+-selective channel, likely αβγ-ENaC. These epithelial cells therefore absorb only Na+ via the amiloride-sensitive pathway and do not provide a parasensory K+ efflux from the canals via this pathway. The results further provide caution to the culture of epithelial cells on impermeable surfaces.

Vitamin D upregulates expression of ECaC1 mRNA in semicircular canal

The low luminal Ca 2+ concentration of mammalian endolymph in the vestibular labyrinth is required for normal balance. We found transcripts in primary cultures of semicircular canal duct (SCCD) epithelial cells from neonatal rats representing a complete transport system for transepithelial absorption of Ca 2+ that is comprised of the epithelial Ca 2+ channels ECaC1 (CaT2, TRPV5) and ECaC2 (CaT1, TRPV6), calbindin (calbindin-D9k, calbindin-D28k), Na +/Ca 2+ exchanger (NCX1, NCX2, and NCX3), and plasma membrane Ca 2+-ATPase (PMCA1, PMCA3, and PMCA4) by RT-PCR. Further, vitamin D receptor was also expressed in SCCD and it was found by quantitative RT-PCR that incubation for 24 h with 1,25-dihydroxyvitamin D 3 upregulated the expression of ECaC1, calbindin-D9k, and calbindin-D28k. These observations provide evidence for the first time of an ECaC-based Ca 2+ transport system in SCCD that could maintain the low Ca 2+ concentration in vestibular endolymph.

In Vivo Study of the Electrochemical Composition of Luminal Fluid in the Guinea Pig Endolymphatic Sac

The aim of this study was to investigate the ionic composition (sodium, potassium) of the luminal fluid in the endolymphatic sac and to correlate it with the transepithelial potential. Experiments were performed in guinea pigs using either an intradural posterior fossa approach or a translabyrinthine approach. The endolymphatic sac transepithelial potential (ESP) was measured and the luminal fluid was sampled. The sodium, potassium and protein concentrations were determined. The results were: i) the luminal fluid in the endolymphatic sac differs in composition from perilymph, on the one hand, and from both cochlear and vestibular endolymph, on the other hand, indicating that the endolymphatic sac maintains chemical (sodium, potassium) and electrical (ESP) gradients; ii

Cytochemical and patch-clamp studies of calcium influx through voltage-dependent Ca2+ channels in vestibular supporting cells of guinea pigs.

To clarify whether or not vestibular supporting cells have voltage-dependent Ca2+ channels, cytochemical and patch-clamp studies were performed using cells isolated from the ampullae of the semicircular canal of the guinea pig. Image analysis used fura-2 as a Ca(2+)-sensitive fluorescence dye and showed that the intracellular Ca2+ concentration ([Ca2+]i) increased with bath application of high (150 mM)K+, but was unaffected by 80 mM K+. The increase in [Ca2+]i induced by high K+ was completely blocked by 1 microM nifedipine as an L-type Ca2+ channel antagonist. In the patch-clamp whole-cell recording of the isolated supporting cells, the voltage-dependent inward current was induced by a depolarizing pulse lasting 2 s in a high (50 mM) Ca2+ and tetraethylammonium-containing external solution replaced by choline chloride and a Cs(+)-containing internal solution. The inward current was obtained when the membrane was depolarized to -50 mV and maximum current was observed at -10 to +10 mV. This inward current was completely blocked by 1 microM nifedipine. These findings strongly suggest that voltage-dependent Ca2+ channels exist in the vestibular supporting cells and regulate Ca2+ concentration in the vestibular endolymph.

Existence of voltage-dependent Ca2+ channels in vestibular dark cells: cytochemical and whole-cell patch-clamp studies.

To determine whether functional Ca2+ channels are present in vestibular dark cells, changes in intracellular Ca2+ concentration ([Ca2+]i) due to K+ applications were measured using the Ca(2+)-sensitive dye (fura-2) and patchclamp whole-cell recordings were made in dark cells isolated from the ampullae of the semicircular canal of the guinea pig. Exchange of the external solution with a buffer medium containing a high K+ concentration (80 mM K+ or 150 mM K+) caused a concentration-dependent increase in [Ca2+]i in vestibular dark cells. Application of 1 microM nifedipine as a Ca2+ channel antagonist completely blocked the increase in [Ca2+]i. Further treatment with 10 microM BAY K 8644 as a Ca2+ channel agonist caused an increase in [Ca2+]i. In the patch-clamp whole-cell recordings a 1-s depolarizing pulse given into the dark cell in the presence of a high barium concentration (50 mM Ba2+) induced an inward current. In determining the current-voltage relationship, a current was detected at a potential that depolarized at-50 mV and was maximal at +10 mV. This inward current was completely blocked by 1 mM La3+ as a Ca2+ channel antagonist. These findings suggest the presence of voltage-dependent Ca2+ channels in dark cells, which have a presumed function in the regulation of [Ca2+]i in the vestibular endolymph.

Ultrastructure of the vestibular dark cell area in patients with acoustic neurinoma.

The human vestibular dark cell (DC) areas of the utricle and ampulla of the lateral semicircular canal were investigated ultrastructurally in 7 patients with acoustic neurinoma. Two types of DCs mainly constituted the epithelial cells of the DC area. One type of DC had basolateral infoldings that were closely interwoven with the melanocyte processes, and the pinocytotic vesicles were frequently found within the basolateral infoldings and the melanocyte processes. The other type of DC had basolateral infolsings stacked upon each other. The former type of DC was more frequently found than the latter type in the DC area adjacent to the sensory epithelium and in the mid portion of the DC area. However, the latter type of DC was more predominant in the DC area distant from the sensory epithelium. These findings suggest that the activity of DCs is modulated by the melanocytes and that the former type of DC has a more active role in ion and/or fluid transport than the latter type of DC. Thus, the DC area near the sensory epithelium might be more actively engaged in the production and regulation of vestibular endolymph than that distant from the sensory epithelium.

前庭暗細胞におけるカルシウムチャンネルの存在

To determine whether the functional Ca2+ channel exist in the vestibular dark cells, an increase in intracellular Ca2+ concentration ([Ca2+]i) due to high K+ application was measured in dark cells isolated from the ampullae of the semicircular canal of the guinea pig using the Ca2+ sensitive dye, Fura-2, and digital imaging microscopy. When 40 mM K+was added to the bath, there was a rapid increase in [Ca2+]i which returned to the original level after washing. Application of the Ca2+ channel blocker, nifedipine (1 μM) blocked the incraese in [Ca2+]i. These results suggest the existence of voltage-dependent Ca2+ channels in the dark cells, which are presumably involved in the regulation of Ca2+ concentration in vestibular endolymph.

Functional Aspects of the Vestibular Dark Cells in the Guinea Pig: Morphological Investigation Using Ruthenium Red Staining Technique

The functional significance of the vestibular dark cells in the guinea pig was investigated using ruthenium red staining technique. The apical cell surface of the dark cell was covered by the fuzzy layer of the glycocalyx, which could be responsible for water and ion transport across the apical plasma membrane. It has been suggested that the otoconia have been at first demineralized by an unknown mechanism when they are attached to the dark cell surface, and the mucopolysaccharide and mucoproteins in the otoconial matrix are then absorbed by active pinocytosis in the dark cells. These functions may be closely related to the possible regulation of the calcium ion in the vestibular endolymph.

Ultrastructure of Ampullar Dark Cells in the Absence of Circulating Adrenocorticosteroid Hormones

The morphology of rat ampullar dark cells was studied in the absence of circulating adrenal hormones at two time intervals of 8-14 and 22-28 days. Five morphological parameters of dark cells were quantitatively determined: cell area per baseline length, basolateral membrane surface density, basolateral membrane length per baseline length, volume density of intercellular space and volume density of vacuoles. Significant statistical reduction of basolateral membrane length per baseline length was observed after an absence of circulating adrenal hormones for 8-14 days and 22-28 days compared to sham operated animals that served as control. The basolateral membrane surface density decreased significantly in the animals sacrificed 22-28 days following adrenalectomy compared to sham operated animals. Cell area per baseline length decreased and volume density of vacuoles increased at both time intervals following adrenalectomy; however, these differences were statistically insignificant. The volume density of intercellular space between dark cells was practically constant among the groups. Results of this study are similar to those reported for ion-regulating nephron cells, and support the hypothesis that cells involved in microhomeostasis of vestibular endolymph may at least be partly regulated by adrenal hormones.

Cytochemical studies of Ca++-ATPase activity in the vestibular epithelia of the guinea pig

We used ultracytochemistry to examine Ca++-ATPase activity in the vestibular epithelia of the guinea pig. Many reaction products were found along the basolateral plasma membrane of the vestibular dark cell. There were also marked reaction deposits on the apical and lateral cell membranes of the transitional cells, and the utricular and saccular wall cells. Both sensory and supporting cells showed Ca++-ATPase activity along their ciliary membrane and apical-lateral cell surfaces. Our findings indicate that the Ca++-ATPase activity found on the plasma membrane is closely related to Ca++-transport across the plasma membrane. When either Ca++ or ATP was omitted from the incubation medium, enzyme activity (as seen by the staining reaction present) was completely abolished. Our present results suggest that Ca++-ATPase located in the vestibular epithelia plays a significant role in the regulation of the Ca++-concentration in the vestibular endolymph.

Elemental composition of the developing inner ear.

The elemental composition of the inner ear fluid-filled compartments has been analyzed using the x-ray energy dispersive technique (CBA mouse). Special attention has been focused on the maturation of endolymph. A few days before and after birth the relative peak intensity of potassium (RK) in the vestibular endolymphatic compartment was slightly surpassing that in the cochlear part of the labyrinth. From the fourth day after birth (DAB) a rapid increase occurred concerning the RK. The highest RK was found in the endolymphatic space in the basal part of the cochlea followed by that in the vestibular endolymph. The lowest RK was measured in endolymph of the apical part of the cochlea. These obvious differences were abandoned already on the sixth DAB. A mature composition of endolymph was reached on the eighth DAB. The present technique does not allow analyses of differences between cochlear and vestibular endolymph with regard to minimal shifts.