Type 1 allergy-induced endolymphatic hydrops and the suppressive effect of H1-receptor antagonist (olopatadine hydrochloride).

OBJECTIVE: To investigate whether endolymphatic hydrops (EH) is experimentally induced by type 1 (or immediate) hypersensitivity allergic reaction and to investigate the inhibitory action of a histamine H(1)-receptor antagonist (olopatadine hydrochloride [OLO-Hy]) on allergic EH induced by systemic immune challenge with 2,4-dinitrophenylated-Ascaris (DNP-As). METHODS: The experimental animals were actively sensitized with DNP-As twice at a 4-week interval and were provoked by an injection of DNP-BSA including DNP-As 1 week after the second sensitization. The OLO-Hy (+) group received oral administration of OLO-Hy (30 mg/kg) 1 hour before the provocation, whereas the OLO-Hy (-) group received distilled water. The temporal bones in all animals were light microscopically examined to assess the degree of EH quantitatively and the expression of degranulated mast cells in the endolymphatic sac. RESULTS: Endolymphatic hydrops was observed 1, 6, 12, and 24 hours after the last sensitization in the OLO-Hy (-) group but was not observed in the OLO-Hy (+) group. Quantitative analysis of the increase ratios (IRs) of the cross-sectional area of the scala media revealed that the IRs of the OLO-Hy (-) group were significantly greater compared with those of the control group (p < 0.001). There was also a significant difference in the IRs between the OLO-Hy (-) and OLO-Hy (+) groups (p < 0.001). CONCLUSION: The systemic sensitization with DNP-As produced allergy-induced experimental EH by type 1 hypersensitivity allergic reaction, and the development of this EH was prevented by histamine H(1)-receptor antagonists.

Localization of aquaporins in the mouse vestibular end organs.

CONCLUSION: We found that aquaporins (AQPs) in the fluid transporting cells, such as vestibular dark cells and endolymphatic sac epithelial cells, seem to be of importance in fluid transport in the inner ear, while those in the sensory and ganglion cells may play a functional role in sensory cell transduction. OBJECTIVE: Expression of AQPs (0-12) was analyzed in normal mouse vestibular end organs. METHODS: CBA/J mice were used in this study. Localization of AQPs 0-12 in the vestibular end organs and endolymphatic sac was investigated by immunohistochemistry. RESULTS: The AQPs were found abundantly distributed in many structures in the vestibular end organs, i.e. vestibular sensory and supporting cells, vestibular dark cells, vestibular ganglion cells, and the endolymphatic sac.

Localization of aquaporins 1, 2, and 3 and vasopressin type 2 receptor in the mouse inner ear.

CONCLUSION: It is suggested that aquaporins (AQPs) 1, 2, and 3, and vasopressin type 2 receptors (V2Rs) in the fluid transporting cells, such as stria vascularis, vestibular dark and transitional cells, and endolymphatic sac epithelial cells, have an important role in fluid transport in the inner ear, while those in the sensory and ganglion cells may play a functional role in the sensory cell transduction system. OBJECTIVE: To analyze expression of AQP1, AQP2, and AQP3 as well as V2Rs in the normal mouse inner ear. METHODS: CBA/J mice were used in this study. Localization of AQP1, AQP2, AQP3, and V2Rs in the inner ear, i.e. cochlea, vestibular end organs, and endolymphatic sac, was investigated by immunohistochemistry. RESULTS: The results show that AQP1, AQP2, AQP3, and V2Rs are abundantly distributed in many inner ear structures, i.e. stria vascularis, inner and outer hair cells, spiral ganglion cells, vestibular sensory and ganglion cells, vestibular dark and transitional cells, and the endolymphatic sac.

Type 1 allergy-induced endolymphatic hydrops and the suppressive effect of leukotriene receptor antagonist.

OBJECTIVE: To investigate allergic endolymphatic hydrops (EH) and the effect of leukotriene receptor antagonist (LTRA). METHODS: Experiment 1: Thirty-six guinea pigs were actively sensitized with DNP-ascaris twice and were provoked with DNP-bovine serum albumin 1 week after the second sensitization. Alterations in the inner ear were investigated histologically at 1, 12, 24, and 36 hours after the provocation, and changes of the endolymphatic space were quantitatively assessed. The animals in the control group received no sensitization but only distilled water. Experiment 2: Twenty-four guinea pigs were actively sensitized and provoked in the same manner. Animals received oral administration of LTRA 1 hour before the provocation. Alterations in the inner ear were investigated as same manner as in Experiment 1. Experiment 3: Eleven of 19 guinea pigs were actively sensitized and provoked in the same manner. Eight animals in the control group received distilled water. One hour after these procedures, the changes in p-AVP levels were investigated. RESULTS: Experiment 1: EH was observed 12, 24, and 36 hours after the last sensitization. In these groups, their cross-sectional areas of the scala media were significantly larger than that of the control group. Degranulation of mast cells was observed in the endolymphatic sac. Experiment 2: In animal groups with LTRA, EH was not observed at all. Experiment 3: P-AVP levels were significantly elevated in animals with the sensitization. CONCLUSION: The sensitization with DNP-Ascaris produced allergic EH and elevation of p-AVP, and allergic EH was inhibited by LTRA.

Endocytosis of cationized ferritin in marginal cells of the stria vascularis is regulated by protein kinase, protein phosphatase, and MEK/ERK and PI3-K signaling pathways.

HYPOTHESIS: The endocytosis of cationized ferritin (CF) via a clathrin-mediated pathway is regulated by a signaling network. BACKGROUND: Marginal cells showed the active endocytosis of CF via a clathrin-mediated pathway. The internalization of receptors through the clathrin-mediated pathway is an important regulatory event in signal transduction. Numerous kinases are involved in endocytosis, and each endocytic route is subjected to high-order regulation by cellular signaling mechanisms. METHODS: CF was infused into the cochlear duct with phorbol 12-myristate 13 acetate, okadaic acid, staurosporin, phenylarsine oxide, PD98059, SB20580 and wortmannin. Endocytic activity was measured at 30 minutes post-infusion by transmission electron microscopy. RESULTS: The endocytosis of CF was stimulated by a protein kinase C activator (phorbol 12-myristate 13 acetate) and a protein kinase A activator (8-bromoadenosine-3', 5'-cyclic monophosphate). It was inhibited by protein phosphatase inhibitors (okadaic acid and phenylarsine oxide), mitogen-activated protein kinase/extracellular signal-related kinase inhibitors (PD98059 and SB20580), and a phosphatidylinositol 3-kinase inhibitor (wortmannin). CONCLUSION: Our previous study showed the endocytosis of microperoxidase to be strongly dependent on protein kinase C, protein phosphatase, extracellular signal-related kinase, and phosphatidylinositol 3-kinase signaling networks but not on protein kinase A and mitogen-activated protein kinase signaling networks. The present study indicated that the signaling cascade regulating CF's internalization differed from the cascade for microperoxidase's endocytosis.

Endocytosis of microperoxidase in marginal cells is mainly regulated by RhoA signaling cascade, but not by Rho-associated protein kinase, myosin light-chain kinase and myosin phosphatase.

Endocytosis plays an important role in cell function and the activation and propagation of signaling pathways. Signaling occurs on endocytic pathways and signaling endosomes, and endocytosis is subjected to high-order regulation by cellular signaling mechanisms. Marginal cells showed active endocytosis of microperoxidase (MPO) via the clathrin-independent pathway. We examined the signaling pathway that regulates MPO endocytosis in marginal cells using specific inhibitors and activators of signaling molecules. The results showed that pertussis toxin - which inhibits the ribosylation of G-protein-coupled receptor - did not affect MPO endocytosis, but Clostridium botulinum C3 toxin - which induces RhoA inactivation resulting in extracellular-signal-related kinase inactivation - inhibited MPO endocytosis. The main endocytotic pathway of MPO did not depend on the Rho-associated protein kinase molecular switch or actin/myosin motor system, but was mainly regulated by the RhoA signaling cascade.

Hormonal aspects of Ménière's disease on the basis of clinical and experimental studies.

CONCLUSIONS: Endolymph homeostasis is thought to be mediated by the vasopressin-aquaporin-2 (VP-AQP2) system in the inner ear. Endolymphatic hydrops, the morphological characteristics of Ménière's disease (MD), seems to reflect the malregulation of the VP-AQP2 system in inner ear fluid. The elevation of plasma vasopressin (p-VP) level, which is often observed in MD and its related diseases, might be one of the causative factors underlying these diseases. PURPOSE OF REVIEW: Review of the role of the VP-AQP2 system in the inner ear fluid homeostasis and in the formation and development of endolymphatic hydrops. RECENT CLINICAL AND EXPERIMENTAL FINDINGS: A clinical survey has revealed that the p-VP level is often elevated in MD and its related diseases and that the increase in the p-VP level was closely linked to vertigo attacks in MD. Experimental studies have revealed that proteins and mRNAs of aquaporin-2 and vasopressin type 2 receptor were expressed in the stria vascularis of the cochlea and the epithelium of the endolymphatic sac, and that the volume of the endolymphatic compartment was mediated by the activity of the VP-AQP2 system in the inner ear.

Narrow-band evoked oto-acoustic emission from ears with normal and pathologic conditions.

CONCLUSION: Evoked oto-acoustic emission (EOAE), in particular the slow component, is fragile with the inner ear lesions and is apt to disappear in impaired ears. This presence is thought to mean that inner ear is not badly damaged, and that the presence of EOAEs in early stage sudden deafness carries a good prognosis. Narrow-band EOAE analysis would open a potentially promising way to manage sensorineural deafness. OBJECTIVE: The aim of present study was to evaluate the characteristics of EOAEs from pathologic ears by a narrow-band EOAE analysis, which allowed us to investigate amplitude, frequency content and latency of EOAEs simultaneously and also to easily detect weak echoes in cases with inner ear lesions. MATERIALS AND METHODS: EOAEs were analyzed by investigating narrow-band frequency contents of EOAEs, filtered by a 100-Hz step of pass bandwidth in frequency regions from 1.0 to 2.0 kHz, and by 500 Hz of pass bandwidth in the frequency ranges of 0.5-1.0 and 2.0-5.0 kHz. EOAE testing was performed in 40 normal ears and 111 ears with pathologic disorders, including sudden deafness, Ménière's disease and surgically proven acoustic neurinomas. Spontaneous oto-acoustic emission was investigated in some cases. In acoustic neurinoma, especially computed tomography scan and magnetic resonance imaging tests were performed to assess the tumor size. RESULTS: (1) Narrow-band EOAE analysis revealed that EOAEs from normal ears were composed of two main echo trains and several sub-echoes. The main echo trains were divided into a fast component with a short latency of <10 ms and a slow component with a long latency of >10 ms. (2) EOAEs could often be detected from ears with moderate to severe hearing loss >45 dB HL in early stage sudden deafness. The prognosis of sudden deafness was good in cases where both a fast component and slow component were detected in the acute stage within 2 weeks after the deafness onset, and was pessimistic, when either or both of them failed to recover. (3) In Ménière's disease, EOAE was found in 6 (40%) of 15 cases with hearing loss >50 dB, and detected in 54 (90%) of 60 cases with slight to moderate deafness <50 dB HL. Echo duration tended to become shorter, and the slow component decreased in amplitude even in ears with slight deafness <30 dB. The detection threshold of the slow component was also elevated. In ears with more advanced deafness, the slow component disappeared and only the fast component with short latency persisted. Ultimately, the fast component also faded out if the hearing was severely impaired. (4) EOAEs were detectable in 20 (95.2%) of 21 ears with surgically proven acoustic neurinoma, 16 of which had both the slow and fast components. The echo pattern of acoustic neurinoma was basically similar to that of normal ears, but the detection threshold was elevated to a varying degree, although there were some cases with much better detection threshold as compared with severe deafness.

Modification of surgical procedures of type 1 tympanoplasty for non-cholesteatomatous chronic otitis media.

OBJECTIVE: To introduce our modified procedures of type 1 tympanoplasty and to assess their efficacy. METHODS: We modified the surgical procedures of type 1 tympanoplasty and have used these procedures since September 1999. The modified points are enlargement of the facial recess approach, no elevation of the posterior meatal skin and the tympanic annulus, and endoaural repair of tympanic membrane perforation. 51 patients with simple chronic otitis media have undergone this modified type 1 tympanoplasty. Postoperative hearing was evaluated according to the criteria proposed by the Otological Society of Japan. RESULTS: The average follow-up period was 15 months (range 6-35). The hearing result was considered successful when the postoperative hearing level satisfied with at least one of three conditions as follows: (1) air-bone gap <15 dB, (2) hearing gain >15 dB, or (3) hearing level >30 dB. The success rate was 94.1%. The average postoperative air-bone gap, hearing gain and hearing level were 3.9, 10.0 and 29.3 dB, respectively. CONCLUSION: Our modified tympanoplasty is useful to achieve better postoperative hearing results.

Expression of aquaporin1, 3, and 4, NKCC1, and NKCC2 in the human endolymphatic sac.

OBJECTIVE: To locate aquaporin (AQP) 1, 3, and 4, Na-K-2Cl cotransporter (NKCC) 1 and 2 in the human endolymphatic sac (ES). METHODS: A sample of human ES was harvested during the removal of vestibular schwannoma via the translabyrinthine approach. The sample was immediately fixed in 4% paraformaldehyde and embedded in OCT compound. Immunohistochemistry was performed with AQP1, 3, and 4, NKCC1, and NKCC2 polyclonal antibodies. RESULTS: AQP1, AQP3, and NKCC2 were strongly expressed in the epithelial layer of the ES. AQP4 and NKCC1 were weakly expressed in the epithelial layer of the ES. CONCLUSIONS: As it is impossible to perform quantitative analysis based on the fluorescence intensity of each immunoreactivity, we have presented the existence of AQP1, 3, and 4, NKCC1, and NKCC2 in the ES. The expression of NKCC1 and 2 indicated that the ES may have both secretory and adsorptive functions to maintain the homeostasis of endolymph.

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.

Expression and immunolocalization of aquaporin-6 (Aqp6) in the rat inner ear.

CONCLUSION: Since aquaporin-6 (Aqp6) protein was located in the membrane of intracellular vesicles of the stria vascularis, endolymphatic sac, and vestibule, Aqp6 might be involved in some distinct physiological function of acid-base metabolism and water balance in endolymphatic fluid homeostasis. However, its lack of expression on the plasma membrane indicates that Aqp6 does not have a direct role in water flux via the plasma membrane. OBJECTIVE: To evaluate the expression and immunolocalization of Aqp6 in the rat inner ear. MATERIALS AND METHODS: Wistar rats were used. Aqp6 mRNA expression in the rat inner ear was investigated in the vestibulum as well as in the cochlea and endolymphatic sac using the reverse transcription-polymerase chain reaction (RT-PCR) method, and detailed immunolocalization of Aqp6 in the rat inner ear was investigated using immunohistochemical methods including immunofluorescence microscopy and immunoelectron microscopy. RESULTS: We obtained novel data showing that not just Aqp6 mRNA but also Aqp6 protein is expressed in the cochlea, endolymphatic sac, and vestibule. Immunoelectron microscopic studies revealed that the immunolabelled gold was diffusely seen in the intracellular area of the stria vascularis, endolymphatic sac, and vestibule, but never in the plasma membranes.

Actin filaments and microtubules regulate endocytosis in marginal cells of the stria vascularis.

CONCLUSION: Cationized ferritin (CF) was internalized via clathrin-mediated endocytosis. This process depends on clathrin, actin filaments, and microtubules. Microperoxidase (MPO) was internalized via a clathrin- and caveolin-independent endocytic pathway, which was partially dependent on microtubules but independent of clathrin and actin filaments. OBJECTIVE: We investigated the role of actin filaments and microtubules in the transport of endocytic carrier vesicles (ECVs) from the plasma membrane to the early sorting endosomes, using CF and MPO as tracers. MATERIALS AND METHODS: Fifty-five guinea pigs were used. The animals were divided into a CF endocytosis group and an MPO endocytosis group. These groups consisted of control, nocodazole-treated, cytochalasin (Cyt D)-treated, Cyt D + nocodazole-treated, and geldanamycin-treated subgroups. RESULTS: For CF endocytosis, the following results were obtained. In the nocodazole experiment, in which microtubules were disrupted to form monomeric tubulin, the number of ECVs loaded with CF was greatly decreased. In the Cyt D experiment, in which the actin filaments were disrupted to form monomers, the number of ECVs labeled with CF was also greatly decreased. In the geldanamycin experiment, in which clathrin-mediated endocytosis was regulated and actin stress fibers were dissolved, the endocytosis of CF was severely inhibited. For MPO endocytosis, in the nocodazole experiment, the endocytosis of MPO was markedly suppressed.

Plasma antidiuretic hormone in cases with the early onset of profound unilateral deafness.

OBJECTIVE: The p-ADH level in cases of juvenile unilateral profound deafness (JUPD) and the timecourse of the level were examined to investigate whether or not an increase of p-ADH is involved in the development of delayed endolymphatic hydrops (DEH) in JUPD. MATERIALS AND METHODS: In 90 consecutive patients with unilateral profound or total sensorineural deafness with the onset in early childhood, pure-tone audiometric examination and the measurement of p-ADH and plasma osmolality (p-OSM) were followed up once or twice a year as far as possible. At every testing, we performed careful history-taking about episodic vertigo/dizziness, fluctuant hearing loss, and tinnitus in order to find out whether patients had experienced these clinical signs of the development of DEH. RESULTS: Means and standard deviation (S.D.) of p-ADH level and osmolality in all samples tested (n=368) were 7.3+/-7.0 pg/mL (0.7-52.0 pg/mL), and 288.6+/-4.4 mOsm/L (273-306 mOsm/L), respectively. The mean of p-ADH level was much higher than those previously reported in children and adolescents. High levels of p-ADH (over 5.0 pg/mL) were often observed in subjects between 6 and 19 years of age, but not so frequently in subjects of 20 years of age or older. Long-term follow-up of p-ADH levels revealed that DEH frequently developed in cases with persistent elevation of p-ADH. CONCLUSIONS: The elevation of p-ADH is likely to promote the development of DEH in cases of JUPD, although the underlying mechanism remains to be elucidated.

Presence and regulation of epithelial sodium channels in the marginal cells of stria vascularis.

CONCLUSION: This study indicates that epithelial Na(+)-selective channels (ENaC) recycle Na(+) via clathrin-mediated endocytosis in the marginal cells of the stria vascularis and that clathrin-independent endocytosis appeared to be modulated by the amount of Na(+) transported. These results suggest the presence of ENaC in the luminal membrane of marginal cells and that ENaC are an efficient pathway for the uptake of Na(+) from the endolymph. OBJECTIVE: The ENaC found in many transporting epithelia play a key role in the regulation of salts and water homeostasis, cellular pH, cell volume, and cell function. Both biochemical and physiological approaches have been used to identify, characterize, and quantify this important channel, but its location in the marginal cells of the stria vascularis has not been fully clarified. The aim of this study was to determine the localization and regulation of ENaC. MATERIALS AND METHODS: Forty healthy female guinea pigs were used: 20 for the control experiment, 10 for the amiloride experiment, and 10 for the aldosterone experiment. We perfused cationized ferritin (CF) and microperoxidase (MPO) as tracers for clathrin-mediated and clathrin-independent endocytosis, respectively, into the cochlear duct. After 30 min of endolymphatic perfusion, the tissues were fixed and CF- and MPO-loaded endosomes within the marginal cell were observed by transmission electron microscopy. The numbers of CF- and MPO-loaded endosomes were compared between the three groups. RESULTS: In the amiloride group, the numbers of CF- and MPO-loaded endosomes decreased in comparison with the control. In the aldosterone group, the numbers of CF- and MPO-loaded endosomes decreased and increased, respectively. Recently, it has been reported that ENaC are endocytosed via clathrin-mediated endosomes and aldosterone decreases the rate of endocytosis of ENaC. In this study, the results of the aldosterone experiment were consistent with those of recent studies.

Expressions of aquaporin-2, vasopressin type 2 receptor, transient receptor potential channel vanilloid (TRPV)1, and TRPV4 in the human endolymphatic sac.

OBJECTIVE: To localize aquaporin (AQP)2, vasopressin type 2 receptor (V2-R), and transient receptor potential channel vanilloid subfamily 1, 4 (TRPV1, TRPV4) in the human endolymphatic sac (ES). METHODS: Three samples of human ES were sampled during the removal of vestibular schwannoma by way of the translabyrinthine approach. The samples were immediately fixed in 4% paraformaldehyde and embedded in OCT compound; immunohistochemistry was performed with AQP2, V2-R, TRPV1, and TRPV4 polyclonal antibodies. RESULTS: AQP2, V2-R, TRPV1, and TRPV4 proteins were detected in the epithelial layer of the ES but were not observed in connective tissue around the ES. TRPV1 was also expressed in blood vascular endothelial cells of the connective tissue of ES. CONCLUSIONS: AQP2, V2-R, and TRPV4 were expressed in the luminal epithelium of human ES. The same characteristic distribution of water and ion channels is seen in the kidney, where a significant amount of fluid is filtrated and resorbed. ES probably plays an active role in the homeostasis of the endolymph.

Endocytosis of microperoxidase in the marginal cells of stria vascularis.

OBJECTIVE: Endocytosis has been thought to control entry into the cell and play a crucial role in the development, immune response, neurotransmission, intercellular communication, signal transduction, and cellular and organismal homeostasis. We investigated the basic properties of endocytosis in the marginal cells of stria vascularis (SV) to discuss whether marginal cells have a potential to maintain the endolymph homeostasis. METHODS: We perfused microperoxidase (MPO), an endocytosis tracer, into the cochlear duct. After 5-60 min of endolymphatic perfusion, the tissues were fixed and the distribution of MPO within the marginal cell was observed by transmission electron microscopy. RESULTS: Endocytosis started already at 5 min after MPO perfusion. Small MPO-loaded endosomes were observed up to 30 min after MPO perfusion. The small tubulovesicular endosomes and the plasma membrane invagination were not decorated by an electron dense bristle structure. After endocytosis, MPO labeled preendosomes were quickly transported to the large vacuolar endosomes that connected with tubular endosomes. At 60 min after MPO perfusion, MPO-loaded large vesicles that have small vesicles in the lumen were observed. CONCLUSION: The time-course of MPO-loaded endosomes was similar to that of CF-loaded endosomes in the marginal cells of SV. The strial marginal cells have vigorous endocytotic activity both in clathrin-independent and clathrin-dependent endocytosis. This high activity of endocytosis in SV seems to be needed to maintain the homeostasis of endolymph via membranous channels and/or receptors regulations.

A comparison of dehydration effects of V2-antagonist (OPC-31260) on the inner ear between systemic and round window applications.

V2-antagonist (OPC-31260 (OPC)) application to the scala tympani reduced endolymphatic hydrops. In the present study, we investigated whether systemic administration or local infusion via the round window (RW application) of OPC would be more suitable for clinical use. In Experiment 1, the increase ratios of the cross-sectional area of the scala media of experimentally induced endolymphatic hydrops were quantitatively assessed among four groups of non-OPC application, RW application of xanthan gum, systemic application of OPC and RW application of OPC. In Experiment 2, the effects of systemic and RW applications of OPC on plasma vasopressin (p-VP) concentrations and plasma osmolality (p-OSM) were investigated. In Experiment 3, endocochlear DC potential (EP) was measured in guinea pigs with the RW application of OPC. Electron microscopic observations of the stria vascularis and the hair cells were also made. Both systemic and RW applications of OPC significantly reduced endolymphatic hydrops. However, systemic application resulted in the distension of the Reissner's membrane in the non-operated ear, which seemed to be caused by elevated p-VP levels resulting from the systemic application of OPC. In contrast, RW application of OPC produced no apparent toxic effects in the inner ear, as indicated electrophysiological or morphological changes. Thus, drug delivery via the round window is more useful for the clinical application of OPC for medical decompression.