Aldosterone inhibits Dot1l expression in guinea pig cochlea.

BACKGROUND: Aldosterone relieves transcriptional repression of epithelial sodium channel (ENaC) by inhibiting Dot1a and Af9 expression and their interaction with ENaC promoter in various tissues. Expressions of ENaC and Af9 in inner ear have been identified. However, it is not known how Dot1l is regulated by aldosterone in inner ear. METHODS: Twenty-eight adult guinea pigs were randomly divided into the control group and treatment group. Aldosterone 1 mg/kg/d was injected intraperitoneally in the treatment group and saline in the control group for 7 days. Animals were killed 1 month later following auditory brainstem response examination. Histomorphology of cochlea was detected with hematoxylin-eosin staining, and Dot1l expression was examined with immunohistochemistry and Western blot. RESULTS: There was no significant difference in ABR thresholds before and after injection of aldosterone or saline in either group. Endolymphatic hydrops was found in 75% of animals in the treatment group. Dot1l was found in both groups in the stria vascularis, Reissner's membrane, spiral limbus, organ of Corti and spiral ligament. Dot1l expression in the treatment group was decreased by aldosterone. CONCLUSIONS: Dot1l in guinea pig cochlea is inhibited by aldosterone with induction of endolymphatic hydrops. Dot1l may be closely related to endolymph regulation by aldosterone and to pathogenesis of Meniere's disease.

Low-salt diet increases mRNA expression of aldosterone-regulated transporters in the intermediate portion of the endolymphatic sac.

The endolymphatic sac is a small sac-shaped organ at the end of the membranous labyrinth of the inner ear. The endolymphatic sac absorbs the endolymph, in which the ion balance is crucial for inner ear homeostasis. Of the three sections of the endolymphatic sac, the intermediate portion is the center of endolymph absorption, particularly sodium transport, and is thought to be regulated by aldosterone. Disorders of the endolymphatic sac may cause an excess of endolymph (endolymphatic hydrops), a histological observation in Meniere's disease. A low-salt diet is an effective treatment for Meniere's disease, and is based on the assumption that the absorption of endolymph in the endolymphatic sac abates endolymphatic hydrops through a physiological increase in aldosterone level. However, the molecular basis of endolymph absorption in each portion of the endolymphatic sac is largely unknown because of difficulties in gene expression analysis, resulting from its small size and intricate structure. The present study combined reverse transcription-quantitative polymerase chain reaction and laser capture microdissection techniques to analyze the difference of gene expression of the aldosterone-controlled epithelial Na+ channel, thiazide-sensitive Na+-Cl- cotransporter, and Na+, K+-ATPase genes in the three individual portions of the endolymphatic sac in a rat model. A low-salt diet increased the expression of aldosterone-controlled ion transporters, particularly in the intermediate portion of the endolymphatic sac. Our findings will contribute to the understanding of the physiological function of the endolymphatic sac and the pathophysiology of Meniere's disease.

A possible mechanism of the formation of endolymphatic hydrops and its associated inner ear disorders.

The pathology of Meniere's disease (MD) is well established to be endolymphatic hydrops. However, the mechanism underlying deafness and vertigo of MD or idiopathic endolymphatic hydrops is still unknown. In order to evaluate the pathogenesis of deafness and vertigo in MD, it seems to be rational to investigate the interrelationship between hydrops and inner ear disorders using animals with experimentally-induced endolymphatic hydrops. In spite of intense efforts by many researchers, the mechanism of vertiginous attack has been unexplained, because animals with experimental hydrops usually did not show vertiginous attack. Recently, there are two reports to succeed to evoke vertiginous attack in animals with experimental hydrops. In the present paper were first surveyed past proposals about underlying mechanism of the development of hydrops and inner ear disorders associated with hydrops, and were discussed the pathogenetic mechanism of vertiginous attack in hydrops. In conclusion, abrupt development of hydrops was thought to play a pivotal role in the onset of vertiginous seizure.

Endolymphatic hydrops and ionic transporters: genetic and biohumoral aspects.

Ménière's disease (MD) is an inner ear disorder, characterized by a burden of symptoms, probably arising from the interplay of genetic and environmental factors. In this brief review, we consider the role of ion channels and transporters in the pathophysiology of MD, focusing on genetic and biohumoral aspects. Pathophysiological mechanisms related to altered concentrations of ions in the endolymph include altered osmotic pressure leading to hydrops and/or immunomodulatory effects of K+ and Endogenous Ouabain (EO) concentrations in the inner ear. Aquaporins 1-5 (AQPs) have been found in the inner ear; AQP2 is the only isoform controlled by a hormone, namely, vasopressin (antidiuretic hormone, ADH). Genetic studies on AQPs have provided inconclusive results. Recently, two genetic polymorphisms have been associated with MD: rs3746951, a missense variant (Gly180Ser) in the Salt-Inducible Kinase-1 (SIK1) gene and rs487119, an intronic variant of gene SLC8A1 coding for a Na+,Ca++ exchanger (NCX-1). EO is a hormone released by the midbrain and adrenal glands. It controls the constitutive capacity of modulating Na+,K+-ATPase activity. Higher plasma levels of EO have been found in MD subjects compared to a control group.

Effect of electroacupuncture on arginine vasopressin-induced endolymphatic hydrops.

OBJECTIVE: To investigate the influence of electroacupuncture (EA) on experimentally induced endolymphatic hydrops (EH) in guinea pigs, and elucidate the association between the dehydrating effect of EA and changes in stria vascularis ultrastructure and expression of vasopressin type 2 receptor (V2R), cyclic adenosine monophosphate (cAMP), and aquaporin 2 (AQP2) in the endolymphatic sac (ES). METHODS: The EH model was established by intraperitoneal injection of arginine vasopressin (AVP). As a treatment, EA was delivered to Baihui (GV 20) and Tinggong (SI 19) acupoints, once daily for 10 consecutive days. For histomorphological studies, degree of cochlear hydrops was evaluated by hematoxylin-eosin staining, and the ratio of scala media (SM) area to SM + scala vestibuli area was calculated. In mechanical studies, ultrastructural changes in stria vascularis tissue were examined by transmission electron microscopy. In addition, cAMP levels and mRNA expression levels of V2R and AQP2 in the ES were compared among groups. RESULTS: EA treatment significantly reduced cochlear hydrops compared with hydropic guinea pigs (P = 0.015). Furthermore, EA attenuated ultrastructural changes in the stria vascularis tissue following EH, significantly upregulated the expression of V2R (P = 0.016), and attenuated AVP-induced upregulation of both cAMP (P = 0.038) and AQP2 expression (P = 0.017) in the ES. CONCLUSION: Collectively, the results of the present study suggest that the dehydrating effect of EA is associated with improvement of stria vascularis ultrastructure and V2R-cAMP-AQP2 signaling pathway regulation in the ES.

Inner ear pathologies impair sodium-regulated ion transport in Meniere's disease.

Meniere's disease (MD), a syndromal inner ear disease, is commonly associated with a pathological accumulation of endolymphatic fluid in the inner ear, termed "idiopathic" endolymphatic hydrops (iEH). Although numerous precipitating/exacerbating factors have been proposed for MD, its etiology remains elusive. Here, using immunohistochemistry and in situ protein-protein interaction detection assays, we demonstrate mineralocorticoid-controlled sodium transport mechanisms in the epithelium of the extraosseous portion of the endolymphatic sac (eES) in the murine and human inner ears. Histological analysis of the eES in an extensive series of human temporal bones consistently revealed pathological changes in the eES in cases with iEH and a clinical history of MD, but no such changes were found in cases with "secondary" EH due to other otological diseases or in healthy controls. Notably, two etiologically different pathologies-degeneration and developmental hypoplasia-that selectively affect the eES in MD were distinguished. Clinical records from MD cases with degenerative and hypoplastic eES pathology revealed distinct intergroup differences in clinical disease presentation. Overall, we have identified for the first time two inner ear pathologies that are consistently present in MD and can be directly linked to the pathogenesis of EH, and which potentially affect the phenotypical presentation of MD.

[Optimization of delivering minimum Gd-DTPA at the posterior upper point on tympanic medial wall and hT2W-3D-FLAIR sequence for detecting endolymphatic hydrops].

Objective: To optimize delivery of gadolinium-diethylenetriamine pentaacetic acid(Gd-DTPA) at the posterior upper point on tympanic medial wall and heavily T2-weighted 3-dimensional fluid-attenuated inversion recovery (hT2W-3D-FLAIR) sequence, and to implement the technique of detecting endolymphatic hydrops using gadolinium-enhancement MRI. Methods: Thirteen patients with periphery vertigo, who visited Department of Otorhinolaryngology Head and Neck Surgery, Shanghai Changhai Hospital during June and December of 2017, were enrolled in the study.0.10-0.20 ml of Gd-DTPA in various dilutions (10, 20, and 40-fold) were delivered at the posterior upper point on tympanic medial wall using a soft-tipped tympanic suction and drug-spraying needle through an artificially perforated tympanic membrane. Inner ear MRI was performed at 8, 24 h after Gd-DTPA administration using a 3T MR machine in combination with a 20-channel Tim 4G head/neck coil and the sequence of hT2W-3D-FLAIR to detect the gadolinium-enhancement signal within the inner ear and possible endolymphatic hydrops. The scanning time was either 8 min 35 s or 15 min 11 s. Results: Efficient inner ear uptake of Gd-DTPA was detected and induced high signal to noise ratio of MRI in patients receiving targeted delivery of 0.15-0.20 ml of 10-fold diluted contrast agent at the posterior upper point on tympanic medial wall. At 8 h after delivery, significant uptake was detected in the scala tympani and vestibuli of hook region and basal turn of the cochlea, and perilymhatic compartment of the vestibule. At 24 h after delivery, the distribution of Gd-DTPA became homogenous in each turn of the cochlea and perilymphatic compartment of the vestibule. However, obvious individual variance existed in the inner ear uptake when 0.10 ml of 40-fold diluted Gd-DTPA was delivered. Efficient inner ear uptake and high quality images that generated in patients receiving 0.10, 0.15, and 0.20 ml of 20-fold Gd-DTPA demonstrated endolymphatic hydrops with minor individual variance. There was insignificant difference in the enhancement signal of inner ear between 0.15 and 0.10 ml groups when Gd-DTPA was diluted at 20-fold except for the signal of semicircular canal of 0.15 ml group (190.00±53.95 vs 165.50±42.13, t=2.61, P<0.05). There was insignificant difference in the image quality between 8 min 35 s and 15 min 11 s canning time. Various degrees of endolymphatic hydrops were detected in 7 cochleae and 11 vestibule, and both simultaneous cochlear and vestibular endolymphatic hydrops were detected in 4 ears. Cochlear endolymphatic hydrops was detected in all the 3 patients with definite Meniere's disease, and 2 of them had combined cochlear and vestibular endolymphatic hydrops. Endolymphatic hydrops was not detected in patients with possible Meniere's disease nor with symptoms of superior semicircular canal dehiscence. Conclusion: Targeted delivery of 0.10 ml with 20-fold diluted Gd-DTPA (total dosage of 5 µmol) at the posterior upper point on tympanic medial wall in combination with 8 min 35 s scanning time hT2W-3D-FLAIR sequence for inner ear MRI in a 3T MR machine is a clinically practical method to detect endolymphatic hydrops, and reduce the requirement for MRI hardware.

Effect of aldosterone on cochlear Af9 expression and hearing in guinea pig.

CONCLUSIONS: Af9 protein in cochlea may be closely related to endolymph regulation by aldosterone and thus may be involved in pathogenesis of endolymphatic hydrops (EH). OBJECTIVES: EH is the pathological characteristic of Ménière's disease (MD). Aldosterone could induce EH, but its relationship with MD is still controversial. The aim of the present study is to investigate the Af9 protein expression in guinea pig cochlea and regulation of Af9 expression and cochlear function by aldosterone. The role of Af9 in pathogenesis of EH is discussed. METHODS: Thirty guinea pigs were randomly divided into two groups. The treatment group was intraperitoneally injected with aldosterone 0.1 mg/kg/d for 5 days, while the control group was done with saline. Hearing and histomorphology of cochlea were examined. In addition, expression of Af9 protein was studied. RESULTS: The hearing threshold of the treatment group was increased. EH was induced in 73% of guinea pigs in the treatment group, and no EH was found in the control group. Af9 protein was found in spiral limbus, stria vascularis, Reissner's membrane, organ of Corti and spiral ganglion in both groups. Af9 expression in cochlea decreased significantly at protein level after treatment by aldosterone.

[Effects of electroacupuncture on cochlea morphology and expression of aquaporins in guinea pigs with endolymphatic hydrops].

OBJECTIVE: To observe the effects of electroacupuncture (EA) on cochlea morphology and expression of aquaporin 1 (AQP1) in guinea pigs with endolymphatic hydrops, so as to explore the possible mechanism of EA on endolymphatic hydrops. METHODS: Forty guinea pigs were randomly divided into a blank group, a model group, a medication group and an EA group, 10 guinea pigs in each one. Model of endolymphatic hydrops was established by using intraperitoneal injection of aldosterone. Guinea pigs in the blank group and model group were treated with identical immobilization as EA group but no treatment was given; guinea pigs in the medication group were treated with intragastric administration of hydrochlorothiazide at a dose of 5 mg/kg, once a day for consecutive 10 days; guinea pigs in the EA group were treated with' EA at "Baihui" (GV 20) and "Tinggong"(SI 19), once a day for consecutive 10 days. The serum ionic concentration in each group was tested by turbidimetric method; hematoxylin-eosin staining was used to measure the severity of cochlea hydrops; immunohistochemical method was used to observe the expression of AQP1 in the cochlea. RESULTS: (1) There was no endolymphatic hydrops in the blank group, moderate-severe endolymphatic hydrops in the model group and slight endolymphatic hydrops in the EA group and medication group. (2) The concentration of K+ and Ca2+ in the EA group was higher than that in the model group and medication group (all P<0. 01); the concentration of Na+ was lower than that in the model group (P< 0. 01) but higher than that in the medication group (P<0. 01); the concentration of Cl- was higher than that in the medication group (P<0. 01), but not significantly different from the model group (P>0. 05). (3) The ratio of expression area of AQP1 in the model group was lower than that in the blank group (P<0. 01); the ratio of expression area of AQP1 in the EA group was higher than that in the model group (P<0. 01), and lower than that in the medication group without significant difference (P>0. 05). CONCLUSION: EA could relieve the endolymphatic hydrops in guinea pigs; the mechanism is likely to be related with up-regulating the expression of AQP1 in cochlea and ion concentration might be an important factor involved.

Developmental changes of ENaC expression and function in the inner ear of pendrin knock-out mice as a perspective on the development of endolymphatic hydrops.

Pendrin mutations cause enlarged vestibular aqueducts and various degrees of sensorineural hearing loss. The selective abolition of pendrin causes dilation of the membranous labyrinth known as endolymphatic hydrops, loss of the endocochlear potential, and consequently loss of hearing function. Because Na+ transport is one of the most important driving forces for fluid transport, the epithelial Na+ channel (ENaC) is believed to play an important role in fluid volume regulation in the inner ear. Therefore, the dysfunction of Na+ transport through ENaC by the acidification of endolymph in Pendred syndrome is one of the potential causes of endolymphatic hydrops. We investigated the changes of ENaC expression and function during the development of the pendrin knock-out mouse. In the cochlea, the expression of ß and γENaC was significantly increased at P56 in Pds-/- mice compared with Pds+/+ mice. In the vestibule, the expression of ßENaC was significantly increased at P56, and γENaC expression significantly increased from P6 to P56 in Pds-/- mice. The ENaC-dependent trans-epithelial current was not significantly different between Pds+/+ and Pds-/- mice in Reissner's membrane or the saccular extramacular roof epithelium at P0, but the current was significantly increased in Pds-/- mice at P56 compared with Pds+/+ mice. These findings indicate that the expression and function of ENaC were enhanced in Pds-/- mice after the development of endolymphatic hydrops as a compensatory mechanism. This result provides insight into the role of Na+ transport in the development and regulation of endolymphatic hydrops due to pendrin mutations.

[Effects of intratympanic injection of dexamethasone on endolymphatic hydrops and changes in guinea pigs inner ear CFTR expression].

OBJECTIVE: To observe the effect of tympanic injection of dexamethasone in the guinea pig endolymphatic hydrops and the change CFTR expression, to explore the effect of glucocorticoid treatment endolymphatic and its possible mechanism. METHOD: Thirty guinea pigs were divided into three groups: hormone group, water group, control group. The animals(hormone group, water group) in study were injected DDAVP 4 µg/kg in the first 7 d, and will increase to 6 µg/kg in the second 3 d. The control group was given normal saline, continuous 10 d. After twelfth days, the hormone group transtympanic injection of dexamethasone (5 mg/ml, 0.5 ml), and water group, control group transtympanic given normal saline (0.5 ml), continuous injection 5 d. Using immuno- histochemistry and Western blot to detect the cystic fibrosis transmembrane conductance regulator cochlear factor (CFTR) expression. RESULT: The water group ABR thresholds was significantly higher than that before the experiment (P < 0.01), and the water group was significantly higher than the rest of the groups (P < 0.01); Hormone group compared with the control group increased threshold value (P < 0.05). The control group had no endolym- phatic hydrops, the water group showed varying degrees of endolymphatic hydrops, cochlear duct and vestibular plus cochlear duct area ratio compared with the control group, hormone group was significantly higher (P < 0.01). hormone group area ratio was higher than the control group (P < 0.05). CFTR was primarily expressed in the stria vascularis, Corti's, spiral ligament, basilar membrane, cochlear ganglion,etc . The expression of CFTR in the water group was increased than that in the control group, and the hormone group (P < 0.01); the expression of hormone group increased compared with the control group (P < 0.05). CONCLUSION: Tympanic injection of dexa- methasone can alleviate the desmopressin acetatein guinea pigs caused by membranous labyrinth, and the improve- ment of the hearing; Tympanic injection of dexamethasone can make the endolymphatic hydrops cochlea of guinea pigs decreased CFTR expression, indicating that the expression and possible mechanisms of CFTR intratympanic steroids reduce endolymphatic hydrops changes.

The predominant vestibular uptake of gadolinium through the oval window pathway is compromised by endolymphatic hydrops in Ménière's disease.

OBJECTIVE: Intratympanic administration of gentamicin is becoming a common therapy to control vertigo in Ménière's disease (MD). Drug entry through an oval window pathway was recently demonstrated in rats. The present study aimed to evaluate the permeability of the oval window to gadolinium in MD patients. METHODS: Eight patients with MD and other inner ear diseases received a transtympanic injection of 5-fold diluted gadopentetate dimeglumine (Gd-DTPA). Three-dimensional fluid-attenuated inversion-recovery (3D-FLAIR) and 3D inversion recovery turbo spin echo with real reconstruction (3D-real IR) imaging was performed using a 3-Tesla MRI scanner at 3, 6, 12, and 24 h after injection. The extent of vestibular endolymphatic hydrops (EH) was determined, and the dynamics of the signal changes in the vestibule and the cochlear basal turn were evaluated. RESULTS: Transtympanically injected Gd-DTPA entered the cochlea of all 8 of the patients and entered the vestibule of 7 of the 8 patients. EH was demonstrated in 2 patients with MD and the patient with idiopathic sensorineural hearing loss or idiopathic vertigo. In the patients lacking EH, the vestibules exhibited more efficient uptake of Gd-DTPA than did the cochleae. In the patients with vestibular EH, the Gd-DTPA signal in the vestibule was less intense than that in the cochlear basal turn. CONCLUSION: In patients with inner ear diseases, the vestibular distribution of Gd-DTPA was compromised by vestibular EH, suggesting oval window passage of Gd-DTPA is reduced in the presence of EH.

Characterization of neuronal cell death in the spiral ganglia of a mouse model of endolymphatic hydrops.

HYPOTHESIS: Spiral ganglion neurons (SGN) in the Phex male mouse, a murine model of postnatal endolymphatic hydrops (ELH) undergo progressive deterioration reminiscent of human and other animal models of ELH with features suggesting apoptosis as an important mechanism. BACKGROUND: Histologic analysis of the mutant's cochlea demonstrates ELH by postnatal Day (P) 21 and SGN loss by P90. The SGN loss seems to occur in a consistent topographic pattern beginning at the cochlear apex. METHODS: SGN were counted at P60, P90, and P120. Semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR), quantitative PCR, and immunohistochemical analyses of activated caspase-3, caspase-8, and caspase-9 were performed on cochlear sections obtained from mutants and controls. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling assay (TUNEL) was carried out on 2 mutants and 2 controls. RESULTS: Corrected SGN counts in control mice were greater in the apical turn of the cochleae at P90 and P120, respectively (p < 0.01). Increased expression of activated caspase-3, caspase-8, and caspase-9 was seen in the mutant. At later time points, activated caspase expression gradually declined in the apical turns and increased in basal turns of the cochlea. Quantitative and semiquantitative PCR analysis confirmed increased expression of caspase-3, caspase-8, and caspase-9 at P21 and P40. TUNEL staining demonstrated apoptosis at P90 in the apical and basal turns of the mutant cochleae. CONCLUSION: SGN degeneration in the Phex /Y mouse seems to mimic patterns observed in other animals with ELH. Apoptosis plays an important role in the degeneration of the SGN in the Phex male mouse.

Expression of osmotic stress protein 94 in murine endolymphatic hydrops model.

CONCLUSION: The up-regulation of osmotic stress protein 94 (OSP94) in the murine endolymphatic hydrops (EH) models suggests that OSP94 might be involved in cellular adaptation in response to ionic and osmotic stress in the murine inner ear. OBJECTIVES: The purpose of the present study was to investigate the expression of OSP94 in cochlear tissues of the murine EH models and control animals. METHODS: Nine adult BALB/c mice were treated with both intratympanic injection of lipopolysaccharide and intraperitoneal administration of aldosterone to induce EH. Nine mice were used as control animals. The expression level of OSP94 in the EH and control groups was compared using immunohistochemistry and real-time RT-PCR. RESULTS: Immunohistochemical staining of tissues in the EH group showed an up-regulation of OSP94 expression in the cochlea, especially in the stria vascularis and Reissner's membrane. Quantitative real-time PCR analysis also showed that transcription of the OSP94 gene in the cochlea was significantly up-regulated in the EH group.

Plasma vasopressin and V2 receptor in the endolymphatic sac in patients with delayed endolymphatic hydrops.

OBJECTIVE: There are some kinds of sicknesses provoked by inadequate adaptation to physical and/or psychogenic stress in daily life. Delayed endolymphatic hydrops (DEH) is an inner ear disease like Ménière's disease (MD) characterized by episodic vertigo in the setting of preexisting unilateral deafness that especially occurs in civilized people with a stressful lifestyle. Its otopathologic finding was demonstrated to be inner ear endolymphatic hydrops through a temporal bone study in 1976, as in the case with MD in 1938. To elucidate the relationship between stress and the inner ear, we examined the plasma antidiuretic stress hormone vasopressin (pAVP) and its type 2 receptor (V2R) expression in the endolymphatic sac in patients with DEH. STUDY DESIGN: A prospective molecular biological study. METHODS: Between 1998 and 2007, we enrolled 20 patients with ipsilateral DEH to examine their pAVP during remission from vertigo attacks. Plasma vasopressin was also examined in 87 patients with unilateral MD and 30 control patients with chronic otitis media. Using the real-time polymerase chain reaction method with tissue samples obtained during surgery, we examined V2R mRNA expression in the endolymphatic sac in 6 patients with ipsilateral DEH, 9 patients with unilateral MD, and 6 control patients with acoustic neuroma. RESULTS: Plasma vasopressin (1.5 times versus controls; unpaired t test, p = 0.140) and V2R mRNA expression in the endolymphatic sac (35.8 times versus controls; unpaired t test, p = 0.002) were higher in patients with DEH compared with those with acoustic neuroma. There were no significant differences in pAVP or V2R expression in the endolymphatic sac between DEH and MD. Patients with DEH showed a significantly negative correlation between pAVP and V2R (Pearson test, r = -0.92, p = 0.009) as in those with MD (Pearson test, r = -0.68, p = 0.043). CONCLUSION: Civilized people are frequently exposed to stress in their daily life, and pAVP can easily become elevated at any time. Therefore, a negative feedback system between pAVP and V2R in the endolymphatic sac may function for inner ear fluid homeostasis against stress-induced increases in pAVP. For the pathogenesis of endolymphatic hydrops resulting in vertigo attacks in patients with DEH as well as MD, pAVP may represent a matter of consequence, but V2R overexpression in the endolymphatic sac could be much more essential as a basis for these diseases.

Individual differences in the permeability of the round window: evaluating the movement of intratympanic gadolinium into the inner ear.

OBJECTIVE: Many recent studies have reported on intratympanic gentamicin therapy for the treatment of intractable Ménière's disease. Intratympanic administration of steroids has also been used to treat sudden sensorineural hearing loss. These intratympanic drug therapies are based on the assumption that the drug administered intratympanically enters the inner ear through the round window membrane. We used magnetic resonance imaging (MRI) to evaluate whether and how intratympanically administered gadolinium (Gd) enters the inner ear. METHODS: GD hydrate was injected intratympanically through the tympanic membrane using a 23-G needle into 61 ears of 55 patients with inner ear diseases. The injected Gd was diluted 8-fold in saline for injection into 58 ears and 16-fold for 3 ears. Three-dimensional fluid-attenuated inversion recovery (3D-FLAIR) imaging was performed using a 3-Tesla MRI unit 1 day after the intratympanic injection. RESULTS: In 53 of 61 ears, the Gd-containing inner ear was detected well as a high signal on 3D-FLAIR imaging. However, Gd was not visible in 2 ears with Ménière's disease and in 1 ear with profound deafness. The concentration of Gd in the perilymph was lower in 4 ears with Ménière's disease and 1 ear with delayed endolymphatic hydrops than after intratympanic administration of the 16-fold Gd dilution. CONCLUSION: Round window permeability was absent in 5% of ears, and 13% of ears had poor round window permeability. These results should be considered when planning intratympanic drug administration therapy to treat inner ear diseases.

Decompression effects of erythritol on endolymphatic hydrops.

OBJECTIVE: The effect of the uptake of erythritol with and without the addition of pectin on fecal condition, p-OSM and p-AVP levels, and the endolymphatic volume was investigated to consider the possibility that erythritol is applicable as a therapeutic agent for Meniere's disease. MATERIALS AND METHODS: Two experiments were performed using 100 female Hartley guinea pigs. Experiment 1 was designed to morphologically investigate the influence of the uptake of erythritol with or without the addition of pectin on the endolymphatic volume. Experiment 2 was designed to investigate changes in p-OSM and p-AVP levels after the uptake of erythritol with or without the addition of pectin. RESULTS: (1) Endolymphatic hydrops significantly decreased after the uptake of a mixture of erythritol and pectin, but did not decrease after the uptake of pectin or erythritol (p<0.001). (2) The fecal condition was muddy in all animals with the uptake of erythritol alone, but muddy or very soft feces were not observed in animals with a mixture of pectin and erythritol. p-AVP and p-OSM levels were significantly elevated in animals with the uptake of erythritol alone or a mixture of erythritol and pectin. Notably, the increase in p-AVP and p-OSM levels was significantly more evident in animals with the uptake of erythritol alone (one-way ANOVA, p<0.001). CONCLUSIONS: The addition of pectin almost completely suppressed erythritol-induced diarrhea. Consequently, the secondary elevation of p-AVP and p-OSM due to diarrhea was also reduced. The uptake of a mixture of erythritol and pectin markedly decompressed endolymphatic hydrops, although the uptake of erythritol alone did not. The difference of the decompression effect between animal groups with the uptake of erythritol alone and a mixture of erythritol and pectin seemed to be attributable to the difference of p-AVP levels due to diarrheal state.

Aquaporins as potential drug targets for Meniere's disease and its related diseases.

The homeostasis of water in the inner ear is essential for maintaining function of hearing and equilibrium. Since the discovery of aquaporin water channels, it has become clear that these channels play a crucial role in inner ear fluid homeostasis. Indeed, proteins or mRNAs of AQP1, AQP2, AQP3, AQP4, AQP5, AQP6, AQP7 and AQP9 are expressed in the inner ear. Many of them are expressed mainly in the stria vascularis and the endolymphatic sac, which are the main sites of secretion and/or absorption of endolymph. Vasopressin type2 receptor is also expressed there. Water homeostasis of the inner ear is regulated in part via the arginine vasopressin-AQP2 system in the same fashion as in the kidney, and endolymphatic hydrops, a morphological characteristic of Meniere's disease, is thought to be caused by mal-regulation of this system. Therefore, aquaporins appear to be important for the development of novel drug therapies for Meniere's disease and related disorders.

[Effect of vasopressin on aquaporin 7 expression in rat inner ear].

OBJECTIVE: To study the effect of vasopressin on aquaporin 7 (AQP7) expression in rat inner ear and reveal the possible role of aquaporins in the formation of endolymphatic hydrops induced by vasopressin. METHODS: Wistar rats were intraperitoneally injected with 50 microg/kg arginine vasopressin once a day for one week. Differentially expressed genes of aquaporins induced by vasopressin injection in rat inner ear were filtered by cDNA microarray. The changes of mRNA expression level of AQP7 in inner ear of rats treated with vasopressin injection were measured by RT-PCR. RESULTS: Differentially expressed gene AQP7 of aquaporins induced by vasopressin injection was screened out in rat inner ear. The expression level of AQP7 mRNA in inner ear of rats treated with vasopressin injection was significantly lower. CONCLUSION: Vasopressin may down-regulate the expression of AQP7 mRNA in the endolymphatic sac and induce a decreased absorption of endolymph, which decreases the water permeability in the potassium ions recycle pathway in the organ of Corti and disturbs the circulation of endolymph, resulting in endolymphatic hydrops.