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Endolymphatic sac tumors (ELSTs) are rare, locally invasive, vascular tumors of the temporal bone. These lesions are associated with von Hippel-Lindau syndrome but may arise sporadically. Early surgical intervention is recommended to prevent permanent neurologic deficits; however, many ELSTs are unresectable or are subtotally resected due to neurovascular compromise. Chemotherapeutic salvage therapy in trials of neoplasms of associated syndromes has targeted angiogenesis with variable response. We present the case of a sporadic ELST, previously minimally responsive to bevacizumab, treated with pazopanib, a multi-kinase inhibitor and antiangiogenic, with good response. Cases such as our patient may demonstrate the utility of novel antiangiogenics in the treatment of these rare neoplasms, particularly when the tumor is unresectable or necessitates subtotal resection.

Introduction: Middle ear application of gentamicin is a common medical treatment for uncontrolled Ménière's disease. The objective of the study was to evaluate the impact of endolymphatic hydrops on inner ear delivery., Methods: Perilymph gentamicin concentrations and correlation with endolymphatic hydrops in an animal model were assessed. A group of 24 guinea pigs was submitted to surgical obstruction of the endolymphatic sac and duct of the right ear. Gentamicin was applied either to the right ear's round window niche or through a transtympanic injection. Perilymph specimens were collected at different times. Histologic morphometry was used to evaluate both turn-specific and overall hydrops degree., Results: In animals with endolymphatic hydrops, lower concentrations of gentamicin were observed after 20 or 120 minutes of exposure and in both types of administration, when compared to controls. This difference reached statistical significance in the round window niche application group (Mann-Whitney, p = 0,007). A negative correlation between perilymphatic gentamicin concentration and hydrops degree could be observed in both groups, after 120 minutes of exposure (Spearman correlation, round window niche p<0,001; TT p = 0,005)., Conclusions: The study indicates that the endolymphatic hydrops degree has a negative interference on the delivery of gentamicin into the inner ear following middle ear application., Competing Interests: The authors have declared that no competing interests exist.

Objective: Endolymphatic sac decompression surgery (ELSD) may be used to treat patients who have Menière's 's disease refractory to medical therapy. In this study, we investigated whether or not the injection of steroid into the endolymphatic sac at the time of ELSD provides additional benefit to patient outcomes., Study Design: Randomized prospective single-blinded placebo-controlled study., Setting: Tertiary center., Patients: Patients with Menière's disease with poorly controlled vertigo despite medical therapy and serviceable hearing that were offered ELSD., Intervention(s): Patients randomized into two groups, with control group (n = 17) undergone ELSD without steroid injection and experimental group undergone ELSD with steroid injection (n = 18) MAIN OUTCOME MEASURE(S):: Audiogram, dizziness handicap inventory, tinnitus handicap inventory, frequency of vertigo spells, functional level scale, and quality of life were obtained at multiple intervals from preoperatively to 24 months postoperatively., Results: ELSD resulted in a statistically significant improvement in vertigo control whether or not steroid was injected into the endolymphatic sac at the time of surgery. However, no additional benefit was observed with the addition of intra-sac steroid injection. No statistical difference in pure-tone average, tinnitus handicap inventory, dizziness handicap inventory, or quality of life was observed between the steroid and nonsteroid surgical groups up to 24 months postoperatively., Conclusion: ELSD is an effective treatment for Menière's disease refractory to medical therapy; however, the addition of intra-sac steroid injection at the time of surgery does not seem to result in a further improvement in patient outcomes.

Objective: The correlation between the isosorbide-induced decompression effect on the endolymphatic space and plasma osmolality (p-OSM) or plasma arginine vasopressin (p-AVP) was investigated on comparing two different dosages of isosorbide (2.8 and 1.4 g/kg) to elucidate why the decompression effect is delayed with a large dose of isosorbide., Materials and Methods: Two experiments were performed using 80 guinea pigs. Experiment 1 was designed to morphologically investigate the sequential influence of the oral intake of 1.4- and 2.8-g/kg doses of isosorbide on the endolymphatic volume. The animals used were 50 guinea pigs (control: 10, experimental: 40). All animals underwent surgical obliteration of the endolymphatic sac of the left ear. One month after the surgery, control animals were sacrificed 3 hours after the intake of distilled water, and experimental animals were sacrificed 3 and 6 hours after the isosorbide intake. All of the left temporal bone served for the quantitative assessment of changes in the endolymphatic space, and the cross-sectional area of the scala media was measured from the mid-modiolar sections of the cochlea.Experiment 2 was designed to investigate changes in p-OSM and p-AVP levels 3 hours after the oral intake of isosorbide. Animals used were 15 guinea pigs (control: 5, experimental: 10). The control group received the oral administration of distilled water (4 ml/kg), and the experimental animals were subdivided into two groups consisting of 10 animals each by the dosage of isosorbide (1.4 or 2.8 g/kg). All animals were sacrificed for the measurement of p-OSM and p-AVP concentrations 3 hours after the intake of water or 70% isosorbide solution., Results: Morphologically, an isosorbide-induced decompression effect was noted in animals with both 1.4- and 2.8-g/kg doses of isosorbide. According to the regression analysis, however, the volumetric decrease of the endolymphatic space was more evident in cases with the small dose (1.4 g/kg) 3 hours after the intake (analysis of covariance [ANCOVA], p < 0.001). Six hours after, the decompression effect was significantly greater in cases with the large dose (2.8 g/kg) (ANCOVA, p < 0.001).Isosorbide intake caused a rise in p-OSM levels dose-dependently. The Cochran-Cox test revealed that the differences in the mean values among control and isosorbide groups were significant (p < 0.01). Regarding the p-AVP level, a significant increase was evident in cases with the large dose (2.8 g/kg) (p < 0.01, Cochran-Cox test), and not in cases with the small dose (1.4 g/kg)., Conclusion: An isosorbide-induced decompression effect of the endolymphatic space was evident in spite of two different dosages of isosorbide (2.8 and 1.4 g/kg). Three hours after the isosorbide intake, however, the decompression effect was more marked in the group with the small dose (1.4 g/kg). Since significant rises in p-OSM and p-AVP were evident in the group with the large dose, this early rise of p-AVP due to dehydration seems to be the major reason for the delayed decompression effect in cases with a large isosorbide intake.

The endolymphatic sac (ES) is a cystic structure of the inner ear connected to the cochlea and vestibule, which plays a role in regulating ion homeostasis in inner ear fluid. Disruption of ion homeostasis can cause inner ear disorders with hearing loss and dizziness, such as Meniere's disease. Herein, we found, for the first time, functional evidence for the involvement of β 1 - and β 2 -adrenergic receptors in apical electrogenic ion transport by human ES epithelium by using electrophysiological/pharmacological and molecular biological methods, which were dependent on K + and Cl - ion transport. The apical electrogenic transport was absent or very weak in ES epithelia of patients with Meniere's disease. These results suggested that adrenergic stimulation via β 1 - and β 2 -adrenergic receptors in the human ES was involved in regulation of inner ear fluid ion homeostasis and impairment of this response could be a pathological mechanism of Meniere's disease., Competing Interests: The authors declare no competing financial interests.

We investigated the influence of vasopressin type 2 receptor antagonist (OPC-41061; Tolvaptan) on experimentally induced endolymphatic hydrops (EH) in guinea pigs. In the first series, the endolymphatic sac (ES) of the left ear of all animals was electrocauterized. Four weeks after surgery, the animals were allocated to four groups: three systemic applications groups (saline, OPC 10 and 100 mg/kg) and a local round window (RW) OPC 1 mg/body application group. We examined the histopathology of the temporal bones and assessed volumetric changes of the endolymphatic space in the cochlea and saccule. In the second series, we investigated the effects of systemic and topical applications of OPC on plasma vasopressin (p-VP) concentrations and plasma osmolality (p-OSM). In the first series, we found that EH was reduced in the OPC 10 mg/kg systemic and OPC RW application groups. In contrast, EH increased in the OPC 100 mg/kg systemic application group. In the second series, neither p-VP levels nor p-OSM were significantly different among the non-OPC, OPC 10 mg/kg systemic, and OPC RW application groups. However, in the OPC 100 mg/kg systemic application group, the p-VP level was significantly higher than that in other groups, and p-OSM was higher than that in the non-OPC group. The systemic application of a low dose of OPC and topical application of OPC resulted in reduced EH in the face of minimal systemic effects (p-VP and p-OSM). These findings suggest that OPC-41061 may be one useful treatment option for EH., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Conclusion: A new murine model of Ménière's disease has been developed, based on long-term administration of vasopressin. Induction of vestibular dysfunction in the present animal model can cause additional stress, by reducing inner ear blood flow. Latanoprost, a selective agonist for the FP prostanoid receptor, may become a new remedy for Ménière's disease., Objective: The purpose of this study was to develop a more suitable animal model, with a closer resemblance to the pathophysiological process in Ménière's disease., Methods: Adult CBA/J or ICR mice were treated by subcutaneous injection of vasopressin for 5 days up to 8 weeks. Morphological analyses were performed of the cochlea, vestibular end organs and endolymphatic sac. The effect of latanoprost on the development of endolymphatic hydrops was also examined., Results: All experimental animals showed mild to moderate endolymphatic hydrops, increasing in severity as the vasopressin treatment was prolonged. Animals treated with vasopressin for 8 weeks showed severe endolymphatic hydrops with partial loss of outer hair cells and spiral ganglion cells. These animals also had a reversible vestibular dysfunction following intratympanic injection of epinephrine. Latanoprost inhibited the development of endolymphatic hydrops caused by vasopressin.

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.

To clarify the role of the endolymphatic sac (ES) in the regulation of endolymphatic pressure, the effects of isoproterenol, a beta-adrenergic receptor agonist, and acetazolamide, a potent carbonic anhydrase inhibitor, both of which decrease ES direct current potential on cochlear hydrostatic pressure, were examined in guinea pigs. When isoproterenol was applied intravenously, hydrostatic pressures of cochlear endolymph and perilymph were significantly increased with no change in endocochlear potential or the hydrostatic pressure of cerebrospinal fluid. Acetazolamide produced no marked change in the hydrostatic pressure of cochlear endolymph. In ears with an obstructed ES, the action of isoproterenol on the hydrostatic pressure of cochlear endolymph and perilymph was suppressed. These results suggest that the ES may regulate the hydrostatic pressure of the endolymphatic system via the action of the agents such as catecholamines on the ES.

Conclusions: That the endolymphatic sac (ES) reacts to changes in inner ear blood flow may be important for homeostasis of the inner ear fluid volume and pressure., Objectives: To elucidate the effect of changes in inner ear blood flow on the ES and to learn more about the volume and pressure regulatory function of the ES., Materials and Methods: Epinephrine or sodium nitroprusside (SNP) was injected into the middle ear cavity of adult CBA/J mice. The ES were analyzed morphologically by light microscopy., Results: Epinephrine reduced the luminal size of the ES leading to an accumulation of intraluminal homogeneous substance. Injection of SNP increased the size of the ES lumen, accompanied by a collapse of the lateral intercellular space (LIS) and dense perisaccular tissue. These changes were almost reversed 4 h after injection.

Objective: Meniere's disease is a common inner ear disease with an incidence of 15 to 50 per 100,000 population. Since Meniere's disease is thought to be triggered by an immune insult to the inner ear, we examined intraendolymphatic sac application of steroids as a new therapeutic strategy for intractable Meniere's disease., Study Design: Prospective randomized controlled study., Methods: Between 1996 and 2005, we enrolled and assigned 197 intractable Meniere's patients to three groups in a randomized controlled trial: Group I (G-I)- patients who underwent endolymphatic sac drainage and steroid-instillation; Group II (G-II)-those who underwent endolymphatic sac drainage without steroid-instillation; and Group III (G-III)-those who declined endolymphatic sac drainage. Definitive spells and hearing in all three groups were determined for 2 to 7 years after treatment., Results: According to the 1995 American Academy of Otolaryngology-Head and Neck Surgery (AAO-HNS) criteria, 2-year results demonstrated that vertigo was completely controlled in 88.0% of patients in G-I (n = 100), 85.1% of patients in G-II (n = 47), and 8.0% in G-III (n = 50). Statistically, G-I = G-II>G-III. Hearing was improved in 49.0% of patients in G-I, 31.9% in G-II, and 6.0% in G-III (G-I>G-II>G-III). Results after 7 years showed that vertigo was completely controlled in 78.8% of patients in G-I, 79.2% in G-II, and 25.0% in G-III (G-I = G-II>G-III). Hearing improved in 36.5% of patients in G-I, 8.3% in G-II, and 0.0% in G-III (G-I>G-II = G-III)., Conclusions: From non-surgical observation in G-III for at least 7 years after treatment, steroids instilled into endolymphatic sac in G-I patients significantly improved hearing in intractable Meniere's patients, more so than endolymphatic sac drainage without steroids in G-II patients.

Conclusion: A new murine model for the study of Ménière's disease has been developed by treatment with both lipopolysaccharide (LPS) and aldosterone. Induction of vestibular dysfunction in the hydropic animal model may entail additional stress such as reduced inner ear blood flow, and sudden acute changes in endolymph volume and/or pressure., Objective: The purpose of this study was to develop a more suitable animal model, showing closer resemblance to the pathophysiological process in Ménière's disease., Materials and Methods: Adult CBA/J mice were treated by intratympanic injection of LPS, intraperitoneal injection of aldosterone, or injection of both LPS and aldosterone. Morphological analyses were performed in the cochlea and endolymphatic sac., Results: All experimental animals showed mild to moderate endolymphatic hydrops. Those treated with both LPS and aldosterone showed reversible vestibular dysfunction after the intratympanic injection of epinephrine.

The aquaporins (AQPs) are a family of small transmembrane water transporters. It has recently been revealed that they play a role in regulating homeostasis in the inner ear fluids. Steroid therapy is usually administered to patients with inner ear disorders; however, the mechanism of steroid effects has not been clearly determined. To elucidate the points of action of steroids in the inner ear, we recently examined the distributions of AQP isoform mRNAs in the rat inner ear and identified AQP1-6 mRNAs in the rat cochlea and AQP1, 3, 4, 5 and 6 mRNAs in the rat endolymphatic sac by means of reverse transcriptase polymerase chain reaction (PCR). In this study, we investigated changes in expression of AQP mRNAs in the rat inner ear after i.p. injections of steroids using real-time quantitative PCR and found that AQP3 mRNA in the endolymphatic sac was significantly upregulated in both dose- and time-dependent manners. This result suggests that steroids may effect water homeostasis in the rat inner ear via AQPs.

The final aim of the present study is to see if the endolymphatic sac is really available as a drug delivery system to have effect on the inner ear organs. In the present study, we examined effects of a single insertion of dexamethasone into the rat unilateral endolymphatic sac on mRNA expression of the inner ear aquaporin (AQP) family, transmembrane water transporters and putative endolymphatic fluid modulators, by means of real-time quantitative PCR. Only AQP-3 mRNA expression in the ipsilateral cochlea was significantly up-regulated in comparison with controls and the up-regulation was demonstrated both in dose-dependent and time-dependent manners. These findings suggest that the intra-endolymphatic sac steroids could make regulatory effects on the inner ear AQP-3 expression via vestibular aqueduct and modulate the homeostasis of endolymphatic fluids, encouraging the possibility that the endolymphatic sac could be a therapeutic window for the inner ear disease.

In the present study, two experiments were performed to investigate the influence of OPC-31260 on experimentally induced endolymphatic hydrops in guinea pigs and the regulation of aquaporin-2 (AQP2) mRNA expression in the rat inner ear. In morphological studies, the increases in the ratios of the length of Reissner's membrane (IR-L) and the cross-sectional area of the scala media (IR-S) were quantitatively assessed among normal guinea pigs (normal ears) and three groups with hydropic ears: hydropic ears with no infusion (non-infusion hydropic ears), hydropic ears with an infusion of physiological saline into the scala tympani (saline-infused hydropic ears) and hydropic ears with infusion of 0.3% OPC-31260 into the scala tympani (OPC-infused hydropic ears). IR-Ls in the experimental groups were markedly larger than in the normal ear group, but there was no significant difference among the groups of non-infusion hydropic ears, saline-infused hydropic ears and OPC-infused hydropic ears. The IR-Ss of non-infusion hydropic ears and saline-infused hydropic ears (48.8-49.3%) were statistically different from that of normal ears (6.5%) (Dunnet multiple comparison test, P<0.01). However, IR-S of the OPC-infused hydropic ears (-14.8%) was significantly smaller than those of non-infusion hydropic ears and saline-infused hydropic ears (one-way ANOVA, P<0.01). In the quantitative polymerase chain reaction study, a comparison of the ratio of AQP2 and beta-actin mRNA (MAQP2/Mbeta-actin) was made between water-injected and OPC-31260-injected rats. An intravenous injection of OPC-31260 resulted in a significant decrease in MAQP2/Mbeta-actin both in the cochlea and in the endolymphatic sac (t-test, P<0.001). These results indicate that water homeostasis in the inner ear is regulated via the vasopressin-AQP2 system, and that the vasopressin type-2 antagonist OPC-31260 is a promising drug in the treatment of Meniere's disease.

Although steroid treatment is generally administered for patients with inner ear disorders, including Meniere's disease, the mechanism via which steroids exert their effects remains to be clarified. The aquaporins (AQPs) are a family of small transmembrane water transporters, and it has recently been revealed that they play a role in regulating homeostasis in the inner ear fluids. In order to elucidate the action points of steroids in the inner ear, we firstly identified AQPI, 2, 3, 4, 5 and 6 mRNAs in the rat cochlea and AQP1, 3, 4, 5 and 6 in the rat endolymphatic sac by means of reverse transcription-polymerase chain reaction. Subsequently, we found that intratympanic injections of steroids upregulated AQPI mRNA of the rat cochlea in a dose-dependent manner. These results suggest that steroids may affect water homeostasis in the rat inner ear mainly via AQP1.

Expression of p-glycoprotein (p-gp) and multidrug resistance protein 1 (MRP1) was detected in the vestibular labyrinth and endolymphatic sac (ES) of the guinea pig by immunohistochemical staining using anti-p-gp monoclonal antibody (mAb) C219 and anti-MRP mAb MRPr1. P-gp was detected in capillary endothelial cells of the crista ampullaris, utricle, saccule and ES. MRP1 was detected in the epithelial lining of the crista ampullaris, utricle, saccule, and epithelial cells of the ES. Since p-gp and MRP1 act as extrusion pumps, they may coordinate with each other in vestibular organs and ES and play an important role in the blood-labyrinth barrier.

Immunohistochemical study for inducible nitric oxide synthase (iNOS or NOS II) in the cochlea of guinea pigs was performed after the injection of keyhole limpet hemocyanin (KLH) into the endolymphatic sac. Morphological changes were observed in the cochlea of all animals after the injection of KLH. Increased iNOS expression was detected in the lateral wall, organ of Corti and ganglion cells. It is known that high levels of nitric oxide can lead to inner ear dysfunction. Our results suggest that iNOS may mediate the inner ear disturbance as seen in endolymphatic hydrops.

To enhance the effect of treatment for intractable Meniere's disease, we exposed the opened endolymphatic sac to high concentrations of steroids. This technique--endolymphatic sac drainage and steroid instillation surgery--involves the application of a mass of prednisolone followed by absorbable gelatin sponges soaked in a high concentration of dexamethasone into a sac lumen opened and expanded with a bundle of absorbable gelatin film. These sponges are also placed around the sac and coated with biochemical adhesive so that the medicine is slowly delivered into the sac over a prolonged period of time by means of a natural sustained-release vehicle. The short-term results (6 to 14 months) in 12 patients with Meniere's disease, including those in stage IV, treated by the above techniques showed that definitive spells were completely controlled in all cases. Hearing was improved, and annoyance due to tinnitus was decreased in all cases except one.

The aim of the present work was to assess the effect of various drugs applied locally on the pH of the luminal fluid (pH(lum)) in guinea pig endolymphatic sac. pH(lum) and transepithelial potential, when measured in vivo by means of double-barrelled pH-sensitive microelectrodes, were 7.06 +/- 0.08 and +6.1 +/- 0.34 mV (mean +/- SE; n = 84), respectively, which is consistent with a net acid secretion in the luminal fluid of the endolymphatic sac. Bafilomycin and acetazolamide increased and decreased, respectively, pH(lum). Amiloride, ethylisopropylamiloride, ouabain, and Schering 28080 had no effect on pH(lum). Results obtained with inhibitors of anionic transport systems were inconclusive; e.g., DIDS reduced pH(lum), whereas neither SITS nor triflocin had any effect. We conclude that bafilomycin-sensitive H(+)-ATPase activity accounts for the transepithelial acid gradient measured in the endolymphatic sac and that intracellular and membrane-bound carbonic anhydrase probably participates in regulating endolymphatic sac pH(lum). The relationship between acid pH, endolymph volume, and Ménière's disease remains to be further investigated.

Morphological changes of the endolymphatic sac were analyzed in guinea pigs following microinjection of artificial endolymph into the cochlea or withdrawal of a quantity of native endolymph. Injections were performed into the second turn of scala media with a micro-pump at a rate of 60-100 nl/min, lasting for a period of 4, 7. 5, 15 or 18 min. In withdrawal experiments, endolymph was aspirated from the second cochlear turn over a period of 8 min. For each procedure the contralateral (non-treated) ear served as a histological control. Following artificial endolymph injections of 7. 5 min or more there was an almost total absence of the normal intraluminal homogeneous substance (HS) on the injected side. Our observations suggest that the disappearance of the HS occurs by both enzymatic and macrophagic activity. After endolymphatic withdrawals the ES was found to contain increased amounts of HS. The results could suggest that the volume of fluid in the ES, and hence the volume of the entire membranous labyrinth, may be regulated by a dynamic relationship between active secretion and enzymatic degradation of a lumen-expanding substance that is intimately related to the intraluminal macrophages. The exact mechanism governing these regulatory systems, and their relationship to ion and water movements across the epithelium of the sac, remain to be elucidated.

Using whole-cell patch-clamp technique and Fura-2 fluorescence measurement, the presence of ATP-activated ion channels and its dependence on intracellular Ca2+ concentration ([Ca2+]i) in the epithelial cells of the endolymphatic sac were investigated. In zero current-clamp configuration, the average resting membrane potential was -66.8+/-1.3 mV (n=18). Application of 30 microM ATP to the bath induced a rapid membrane depolarization by 43.1+/-2.4 mV (n=18). In voltage-clamp configuration, ATP-induced inward current at holding potential (VH) of -60 mV was 169.7+/-6.3 pA (n=18). The amplitude of ATP-induced currents increased in sigmoidal fashion over the concentration range between 0.3 and 300 microM with a Hill coefficient (n) of 1.2 and a dissociation constant (Kd) of 11.7 microM. The potency order of purinergic analogues in ATP-induced current, which was 2MeSATP>ATPgammas>/=ATP>alpha, beta-ATP>ADP=AMP>/=adenosine=UTP, was consistent with the properties of the P2Y receptor. The independence of the reversal potential of the ATP-induced current from Cl- concentration suggests that the current is carried by a cation channel. The relative ionic permeability ratio of the channel modulated by ATP for cations was Ca2+>Na+>Li+>Ba2+>Cs+=K+. ATP (10 microM) increased [Ca2+]i in an external Ca2+-free solution to a lesser degree than that in the external solution containing 1.13 mM CaCl2. ATP-induced increase in [Ca2+]i can be mimicked by application of ionomycin in a Ca2+-free solution. These results indicate that ATP increases [Ca2+]i through the P2Y receptor with a subsequent activation of the non-selective cation channel, and that these effects of ATP are dependent on [Ca2+]i and extracellular Ca2+.

The anti-diuretic hormone vasopressin (AVP) regulates water excretion from the kidney by increasing the water permeability of the collecting duct. AVP binds to V2-receptors and induces the translocation of aquaporin-2 water channels (AQP-2) into the apical plasma membrane of principal cells. By this mechanism AVP controls water reabsorption in the kidney. The effects of AVP on the endolymphatic sac (ES) of the inner ear, which is thought to mediate reabsorption of endolymph, were investigated. Both the V2-receptor and the AQP-2 water channel were found to be expressed in the ES epithelium. In the ES AVP binds to receptors most probably of the V2-subtype. Application of AVP to organotypically cultured ES inhibits membrane turnover in ribosomal-rich cells of the ES epithelia, which is thought to mediate translocation of AQP-2 into the surface membrane. This suggests that AVP has contrasting effects in the inner ear and kidney, which may be physiologically useful for maintaining endolymphatic pressure during severe hypovolemia. Animal experiments show that AVP causes endolymphatic hydrops after systemic application to guinea-pigs, which suggests a causal role for the increased AVP levels found in humans suffering from Ménière's disease.

In Ménière's disease, an inner ear disorder related to an endolymphatic hydrops, an alteration of the functioning of the endolymphatic sac has been proposed. The endolymphatic sac is assumed to be involved in the secretion/resorption of endolymph. The epithelial transport systems have been indirectly studied by the recording of the endolymphatic sac transepithelial potential (ESP) in control conditions and after the local injection of drugs such as diuretics that have been proposed in the treatment of Ménière's disease. The ESP was recorded, in vivo, in guinea pigs up to 150 minutes after the perisaccular injection of 5 microL of a 150 mmol/L (mM) NaCl solution containing various drugs known to inhibit ionic transport systems. The initial ESP was +8.4+/-0.3 mV (mean +/- SEM, n = 78). The basolateral injection of 5 microL of 150 mM NaCl induced an ESP decrease of 64%+/-6.0% (n = 12), 5 minutes after the end of the injection. Then ESP increased, returning to its initial value at 60 minutes and surpassing it at 120 minutes. Diuretics such as acetazolamide (10[-3] mol/L [M]), an inhibitor of carbonic anhydrase, and amiloride (10[-4] M), an inhibitor of Na channel or Na/H exchanger, decreased the ESP recovery. At variance, bumetanide (10[-6] M, 10[-4] M), the Na-K-Cl cotransport inhibitor, and chlorothiazide (10[-4] M), a Na-Cl cotransporter inhibitor, failed to alter the ESP as compared with the control group. Ouabain (10[-3] M), the Na+,K+-adenosine triphosphatase (ATPase) inhibitor, prevented the ESP recovery otherwise observed 60 minutes after the NaCl injection. Bafilomycin A1, the inhibitor of the vacuolar-type H+-ATPase, prevented the recovery of the ESP with a log-dose/effect (10[-5] M, 10[-6] M, 10[-8] M). Disulfonic acid stilbene (DIDS) (10[-4] M), an inhibitor of transporters involving HCO3-, also prevented the ESP recovery. These results suggest that the genesis of the ESP was highly dependent on acid-base transport systems including carbonic anhydrase, a vacuolar-type H+-ATPase, and an anionic transport system blocked by DIDS. Further studies are needed to confirm the alteration of the acid-base balance in this epithelium and its possible involvement in the pathogenesis of Ménière's disease.

The membrane potential (Em) in epithelial cells isolated from the intermediate portion of the endolymphatic sac (ES) of the guinea-pig was recorded using the whole-cell patch-clamp technique. In the steady state the Em was -53.5+/-1.5 mV (n = 74), which is similar to that in epithelial cells of other tissues. Application of 1 MM ouabain induced a depolarization of Em by approximately 10 mV (n = 6), suggesting that an active ion transport process by Na+-K+-ATPase may be present in the ES epithelial membrane. Increasing extracellular K+ concentrations from 5 to 100 mM induced a significant membrane depolarization that was close to the K+ equilibrium potential calculated by the Nernst equation, indicating that K+ may be a predominant permeable ion in the ES epithelial membrane. Total replacement of extracellular Na+ with NMDG+ led to a significant membrane hyperpolarization of 38.7+/-2.5 mV (n = 18), suggesting that Na+ may be another major permeable ion for the ES epithelial membrane. Reducing extracellular Cl- concentrations from 149.3 to 7 mM had no significant effect on Em, indicating that Cl- may be a negligible permeable ion in the ES epithelial membrane.

The effects of isosorbide on distortion-product otoacoustic emissions (DPOAEs) and endocochlear DC potential (EP) were examined in experimentally induced endolymphatic hydropic ears and untreated control ears using 20 albino guinea pigs. DPOAEs and EP in the hydropic ears decreased after obliteration of the endolymphatic duct and sac. The administration of isosorbide restored DPOAEs to the normal level, whereas EP was further reduced. In control ears there was no significant change in DPOAEs and EP after the administration of isosorbide. The results obtained in the present study indicate that the effect of isosorbide on DPOAEs is different from that on EP, and the recovery of DPOAEs in hydropic ears is not a secondary phenomenon due to the recovery of EP.

Endothelin (ET), originally characterized as a 21-residue vasoconstrictor peptide from endothelial cells, has been reported to act as a local hormonal regulator of pressure, fluid, ions, and neurotransduction. Our previous studies suggested an important role of ET-1 in the inner ear. The present study investigated the time kinetics of ET-1 in the epithelium of the endolymphatic sac (ES) of guinea pigs and its relation to the development of endolymphatic hydrops (EH) following locally mounted secondary immune reaction. In the duration between 12 h and day 1, ET-1-like activity completely disappeared from the epithelium of the ES and was associated with the accumulation of inflammatory cells in the ES and a rapid development of EH. On day 7, ET-1-like activity recovered as a consequence of the decrease of inflammatory cells and reduction of EH. These findings suggest that ET-1 may play an important role as one of the regulators maintaining the fluid balance.

The classical guinea pig model for Meniere's disease, in which endolymphatic hydrops was achieved by destruction of the endolymphatic sac and obliteration of the endolymphatic duct, is a non-physiological profound model with shortcomings in relation to Meniere's disease as seen in patients. We developed a more subtle animal model; the two-phase endolymphatic hydrops. This model is based on a combination of chronic endolymphatic sac dysfunction, induced by slight destruction of the most distal part of the endolymphatic sac, and acute stress-induced endolymph production by stimulation of the Na/K-ATPase in the stria vascularis with aldosterone. Light microscopy of the fluid compartments of four groups of cochleas was used to examine them for the presence of endolymphatic hydrops: i) Normal (control) cochleas showed no hydrops; ii) some of the non-operated (no destruction) aldosterone-treated cochleas showed small degrees of hydrops mainly present in the basal turns; iii) mild dissection of the endolymphatic sac without administration of aldosterone produced a hydrops which was mainly present in the cochlear apex; iv) combination of chronic endolymphatic sac dysfunction and acute attacks of endolymph production by aldosterone administration revealed the most severe degrees of hydrops in all cochlear windings, damage to cochlear structures, and cellular disturbances of the epithelial lining of the endolymphatic sac. This new model may represent a more physiologic and dynamic approach to Meniere's disease and may explain the etiology of many symptoms in patients such as the fluctuant nature and the types of sensoneuronal hearing losses.

Recent clinical and laboratory evidence indicates that Meniere's disease is an immune-mediated disease. Dexamethasone perfusion of the inner ear through the round window plus intravenous dexamethasone often will stop the dizzy spells, reduce the fullness and low-frequency tinnitus, and sometimes improve the hearing in patients with Meniere's disease. The dexamethasone must act mostly on the endolymphatic sac and, to a lesser extent, on the stria vascularis and spiral ligament, the known targets of immune response in the inner ear, to reduce the endolymphatic hydrops and restore the fluid dynamics of the endolymph. Despite the good results with streptomycin perfusion, the number of patients with further hearing loss is large, so dexamethasone perfusion with intravenous dexamethasone should be tried first. The initial response to dexamethasone perfusion plus intravenous dexamethasone has been very good, with very little risk of further hearing loss, and it holds great promise for the future.

The present study examined the effect of glycerol administration on 3-fucosyl-N-acetyl-lactosamine (CD15) epitope expression in the endolymphatic sac (ES) of the guinea pig's inner ear. Adult guinea pigs were injected intravenously with glycerol (2 g/kg body wt.). CD15 expression was studied at 80 min up to 5 h after treatment. In untreated animals single cells and cell groups in the ES expressing CD15 epitope intra- and intercellularly were identified by immunohistochemistry to be mainly in the epithelial layer of the rugosal and distal part of the sac. Glycerol administration modulated the expression of CD15 epitope. In the epithelial layer, expression decreased and was nearly depleted after 3 h. After 4 h of glycerol administration, CD15 expression reappeared and reached the comparable level of controls. The numbers of CD15-positive cells in the lumen of the ES increased steadily and arrived at their the highest levels in 2-h specimens. The localization of CD15-epitope expression and its modulation after glycerol administration within the ES implies that this molecule may play a role in re-establishing the sac's normal function. In addition, we speculate that CD15 may be associated with processes of an immune response in the inner ear.

The effects of glycerol were investigated in guinea pigs in which the endolymphatic sac (ES) and duct (ED) had been obliterated unilaterally. Twenty-four hours after obliteration of the right ES and ED, glycerol was injected intraperitoneally. One to two hours after administration of glycerol, collapse or folding of Reissner's membrane was observed in the operated cochlea. No remarkable change was found in the nonoperated ear. The lumen of the ES of the nonoperated ear collapsed with intraluminal stainable substance within 1 h. The volume of the stainable substance increased with time and could still be seen 4 h after the injection of glycerol. Twenty-four hours after the injection of glycerol, the configuration of the ES was normal again. In addition, the animals showed marked unilateral (operated side) vestibular dysfunction after injection of glycerol. These findings suggest that the ES also has a secretory capacity and plays an important role in inner ear fluid volume and pressure regulation.

Endolymphatic sac (ES) blood flow (ESBF) and cochlear blood flow (CBF) were measured in different groups of guinea pigs by laser-Doppler flowmetry (Advance Laser Flowmeter, Model ALF 2100) after the intravenous administration of 70% isosorbide (1.6 ml/kg). The measurements were made under general anesthesia with intraperitoneal pentobarbital sodium. Respiration was controlled by a respirator after tracheotomy, and blood pressure was monitored through the femoral artery (Gould Statham P23 ID Pressure Transducer). For ESBF measurements, a probe was placed on the right ES after entering the posterior cranial fossa via the dorsal approach. For CBF measurements, a probe was placed on the basal turn of the right cochlea via the ventral approach. Isosorbide was administered intravenously through the jugular vein for 60 s. Both ESBF and CBF increased immediately after administration, reached a peak within 3-6 min and decreased gradually to their initial baseline levels in 11-15 min. Both blood flow changes almost always corresponded to systemic blood pressure changes, although a slight delay was observed in blood pressure compared to the blood flow. The magnitude of the CBF response tended to be greater than that of the ESBF response (p < 0.1). This may result from the anatomical differences in the two blood supplies, i.e., from the vertebral artery (CBF) and the external carotid artery (ESBF).

One hundred and sixty-eight mice were injected with hyaluronan (HA) of varying tonicity in the middle ear and compared with 81 control animals to study the effects of hypertonic solutions in the middle ear on inner ear fluid homeostasis and endolymphatic sac (ES) response. Six hours after 1.9% HA and 4, 6 and 8 h after 4% HA deposition there was a significant increase in the proportion of granule-containing light epithelial cells in the ES as compared with controls indicating an enhanced secretory activity from the epithelial lining. Six, 8 and 10 h after deposition of 4% HA, the ES volume was significantly reduced. The secretory activity and the ability of the ES to vary its volume could be specific responses to dehydration of the inner ear resulting from the high colloid osmotic pressure exerted by concentrated HA. The secretion of high-molecular substances into the ES may serve to keep the ES lumen patent and thus ensure a proper ES function and/or to regulate the intralabyrinthine volume and fluid pressure. Whether the findings may have clinical relevance for Ménière's patients remains to be elucidated.

This study, using mice, correlated serum osmotic pressure, experimentally elevated with an intravenous injection of glycerol of varying tonicity (1.3, 2.6 and 5.2 g glycerol/kg body weight) with endolymphatic sac (ES) response. The injection produced a rise in serum osmotic pressure that was measured after 15 min (from 308 +/- 12.9 mosm/kg b.w. to 320 +/- 1.9, 353 +/- 19.3, 427 +/- 9.4 mosm/kg b.w., respectively). Normal levels were noted about 60 min after the injection. The ES was graphically displayed at 0 min, 1, 2 and 4 h using as a basis composite electron micrographs. The ES responded with signs of active secretion and degradation of macromolecular substances which appeared to be related to the rise in serum osmotic pressure. The study showed that the ES responds to glycerol injection with fine structural changes indicative of increased intra-epithelial synthesis of sugar/protein residues which are secreted and degraded within its lumen. The technique used made it possible to characterize the ultrastructural secretory pathways from the epithelial cells into the sac lumen. The results indicate that the ES may sense and respond to dynamic changes of the micro-osmotic environment probably via alterations in inner ear fluid homeostasis.

To create a rational treatment system for Meniere's disease, it is helpful to divide Meniere's disease into five stages according to the signs, symptoms, pathology, and natural history by recalling what is known for certain and what is probably true about Meniere's disease. It is known that Meniere's disease is attributable to endolymphatic hydrops, caused by a small, malfunctioning endolymphatic sac, abnormally placed since birth, and Meniere's disease is therefore a congenital disorder of the endolymphatic sac. It is probably true that there is often an immune, viral, metabolic, or other insult to the inner ear, that disturbs the balance between the cochlea, where endolymph is produced, and the endolymphatic sac, where most of it is absorbed. Using this five-stage classification, it is possible to separate Meniere's disease into the first three stages, for which different but reasonable treatments are possible, and the last two stages, for which no treatments are effective.

The effects of acetazolamide, a carbonic anhydrase inhibitor, on acid-base parameters in the endolymphatic sac of the guinea pig were investigated using ion-selective microelectrodes. Bicarbonate concentration was reduced and pH shifted to being acidic immediately after administration of acetazolamide. Chloride concentration in the endolymphatic sac increased after carbonic anhydrase inhibition. These findings provide evidence of an active function of carbonic anhydrase in the endolymphatic sac and suggest that carbonic anhydrase activity is involved in the formation of endolymphatic sac fluid.

Our recent studies have revealed that catecholamines depress the endolymphatic sac DC potential (ESP) by beta-adrenergic action, and that acetazolamide (ACTZ), a potent carbonic anhydrase inhibitor, decreases the ESP. The interaction of ACTZ and isoproterenol (Iso), a nonselective beta agonist, in their action on the ESP was examined in the guinea pig. Intravenous administration of Iso (6.25 micrograms/kg/min) and ACTZ (10 mg/kg) reduced the ESP amplitude by 38.5 +/- 5.9% (n = 8) and 39.8 +/- 3.7% (n = 8), respectively. Co-administration of both agents reduced the ESP amplitude by 62.3 +/- 3.3% (n = 8). The ESP change induced by co-administration was significantly larger than that by the administration of each agent alone. Co-treatment with Iso and ACTZ at doses producing near-maximum reduction of the ESP depressed almost all parts of oxygen-dependent components of the ESP. The results suggest that Iso and ACTZ decrease the ESP via mechanisms different from each other, and that oxygen-dependent components of the ESP are composed of Iso- and ACTZ-sensitive parts.

This study aimed to investigate the origin of the free cells in the lumen of the endolymphatic sac (ES). Activation of the cells was accomplished through osmotic induction using glycerol. The ES and the perisaccular tissue were analyzed with special reference to the activity of periaqueductal bone marrow cells after different time intervals following the injection of hyperosmotic agents. The results show that the perisaccular or periaqueductal bone marrow space may constitute a source of some of the free cells occurring in the ES. Osmotic challenging of the inner ear may cause activation of the periaqueductal bone marrow, initiating the locomotion and migration of cells (mostly monocytes, neutrophils and eosinophilic leukocytes) along bone marrow sinusoids that frequently anastomose with the ES vessels. The free cells show signs of transepithelial diapedesis and, in the lumen of the ES, cells may develop into phagocytes which initiate the ingestion and degradation of secreted macromolecular aggregates. It is thought that osmotic alterations in the inner ear may give rise to local changes in or around the ES, leading to the chemotactic attraction of bone marrow cells. The results verify the existence of a complex sugar/protein aggregate metabolism over the wall of the ES, which is linked to the turnover of free cells. The findings may indicate that ES macrophages are important in the regulation of inner ear fluid homeostasis.

A time sequence study was performed on experimental animals to investigate long-term effects of intravenously administered glycerol on the epithelial cell activity in the endolymphatic sac (ES) and on the ES volume. Fifteen to 60 min after systemic glycerol administration, the ES volume decreased. During this time, the ES lumen was often obliterated. Subsequently, the lumen dilated. Meanwhile, many light epithelial cells showed granules with floccular and/or lamellar contents. Concomitant deposition of floccular material into the luminal space suggested secretion of macromolecular substances, presumably from these transformed light cells. The number of granule-containing cells was significantly increased 2 h (p < 0.01) and 4 h (p < 0.01) after glycerol administration. The ES was significantly dilated after 4 h (p < 0.01) and 6 h (p < 0.05). Thus systemic alterations in osmotic pressure led to a reversible change in ES volume, with initial collapse followed by dilation and normalization after 8 h. The secretory response of the ES preceded the volume increase. A great variability in ES volume indicated high compliance of this organ system. A secretion/degradation system or turnover of osmotically active macromolecular complexes in the epithelial lining and ES lumen seems to be linked to the ability of the ES to hold fluid volumes within a wide range. This may serve as a micromechanical pressure-volume-regulating device for monitoring endolymph fluid homeostasis.

The effect of angiotensin II (ANG II), known as a potent pressor hormone, on the endolymphatic sac direct current potential (ESP) was examined. Intravenous administration of ANG II produced a reversible decrease in the ESP at doses exceeding 0.1 microgram/kg. The effect of ANG II on the ESP was dose-dependent at doses of less than 3 micrograms/kg with a saturated effect at doses of more than 3 micrograms/kg. The maximum reduction of the ESP produced by ANG II was 33.8 +/- 2.9% (m +/- SE, n = 6) of the original amplitude. The action of ANG II on the ESP was blocked by propranolol (beta-adrenergic antagonist). The mechanism underlying the action of ANG II on the ESP is discussed.

Aminoglycosides were applied to the endolymphatic sac of guinea pigs, and the inner ears were studied by light microscopy. In group 1 gentamicin sulfate application on the lateral surface of the endolymphatic sac produced sensory cell atrophy limited to the macula sacculi. In group 2 gentamicin injection into the sac by extradural and intradural approaches often showed lesions in the maculae sacculi and to the sac from a lateral opening resulted in a similar pattern of degeneration but with less severity in comparison to the injection method. Mild to severe endolymphatic hydrops was sometimes observed. The study shows diffusion of drugs taking place against the longitudinal flow of endolymph. This procedure may be applicable for the treatment of Meniere's disease.

The combination of fluctuant hearing loss, fullness, tinnitus and dizzy spells we call Meniere's disease is thought to be caused by endolymphatic hydrops. Most patients with the clinical picture of Meniere's disease do have endolymphatic hydrops but some patients with endolymphatic hydrops do not have the clinical picture of Meniere's disease. It would appear there is an, as yet unknown, immune-mediated, cause for Meniere's disease, in addition to endolymphatic hydrops, and this immune-mediated cause may aggravate those ears with endolymphatic hydrops. While medical treatment with a low-salt diet, diuretics and steroids are of value in some patients when given early in controlling dizzy spells and improving the hearing, there is usually no real, long-term benefit. Since none of these "shunts" of the sac could remain open for more than a few hours, they could have no more direct benefit than a one-time drainage of endolymph, while doing harm to the fluid absorption, immune response and phagocytosis roles of the endolymphatic sac. The various vestibular neurectomy operations, while usually stopping the dizzy attacks, are both difficult and potentially dangerous, but more important, do nothing for the hearing loss. The one direct attack on the problem, both easy to perform and certain to relieve the dizzy attacks, is to destroy the vestibular receptors with streptomycin. This destructive action on the stereocilia and sensory cells, without damage to the rest of the vestibular system and the cochlea, has been verified by two cat experiments with streptomycin by Norris et al. and Norris & Shea and two guinea pig experiments with gentamicin by Kimura.(ABSTRACT TRUNCATED AT 250 WORDS)

The effect of acetazolamide known as a potent carbonic anhydrase inhibitor of the endolymphatic sac DC potential (ESP) was examined in the guinea pig. Intravenous administration of acetazolamide produced a dose-dependent decrease in ESP amplitude at doses of less than 10 mg/kg. The effect of acetazolamide on ESP was saturated at doses exceeding 10 mg/kg. The maximum reduction of ESP induced by acetazolamide was approximately 50% of the original ESP amplitude. The results suggest that carbonic anhydrase is involved in the generation of ESP.

The effect of aldosterone and canrenoate, known as an aldosterone antagonist, on the DC potential in the endolymphatic sac (ESP) was examined in the guinea pig. Intravenous administration of aldosterone (1 mg/kg) induced no change in the ESP for 60 minutes after the injection. However, canrenoate, an aldosterone antagonist, produced a dose-dependent decrease in the ESP. Pretreatment with aldosterone attenuated the decrease in the ESP by canrenoate. The results suggest the possibility that aldosterone may play a role in the endolymph absorption of the endolymphatic sac.

The etiology underlying endolymphatic hydrops is scarcely understood. It is generally accepted, however, that the primary cause of endolymphatic hydrops is the malfunction and malabsorption of the endolymphatic sac. This has already been proven by animal experiments as well as by histopathological studies of human temporal bone. In this study, we attempted to induce endolymphatic hydrops by utilizing both mechanical and immunological methods. An obliterative procedure was performed on the endolymphatic sac and duct of 14 guinea pigs, 12 of which showed extensive endolymphatic hydrops after a variable period of time. The immunological method employed was to inject the antigen horseradish peroxidase into the endolymphatic sac of 22 guinea pigs that had been systemically sensitized to this antigen. Endolymphatic hydrops was induced in 11 of these subjects. Thus, this study lends further support to the fact that endolymphatic hydrops is caused by malfunction of the endolymphatic sac, resulting from either mechanical obliteration or immunological reaction.

Hyaluronan (HA) 0.5, 1, 1.9 and 4% was instilled into the round window niche in mice. Six hours after the 1.9% HA and 4, 6 and 8 h after the 4% HA deposition there was a significant increase in the proportion of granule-containing light epithelial cells in the endolymphatic sac. This indicated an increase in secretory activity. This activity could be a response to lowered intralabyrinthine pressure due to the osmotic capacity of HA. Secretion of high molecular substances can serve to keep the sac lumen patent and/or to regulate labyrinthine volume and pressure.

The growth rate of calcium carbonate (CaCO3) crystals in the endolymphatic sac was modulated, and morphological changes in this organ were observed by light and electron microscopy. When calcium chloride (CaCl2) was given to the three frog for a short period (3 days to 2 weeks), CaCO3 crystal production was accelerated. Epithelial cells enlarged, their rough endoplasmic reticulum (rER) and Golgi apparatus developed, and dense material increased around CaCO3 crystals and/or in the endolymphatic lumen. In addition, multiluminal endolymphatic chambers appeared in some frogs. On the other hand, as the CaCl2 loading period lengthened and CaCO3 crystal formation decreased or stopped, the epithelial cells became flat and extended with scanty cytoplasm, and the rER and Golgi apparatus decreased in number and size. Furthermore, the amount of dense material around CaCO3 crystals and in the lumen decreased. These findings suggest that the rER, Golgi apparatus and dense material have key roles in the production of CaCO3 crystals.

Since much evidence suggests that the endolymphatic sac is responsible for endolymph resorption, and that the endolymphatic sac lateral intercellular spaces which are lined by the energy-dependent transport complex NA+,K(+)-ATPase are important in this process, we sought to evaluate the effects of dietary salt and a food extract that inhibits the activity of Na+,K(+)-ATPase on lateral intercellular space size. Animals fed this food factor and a high-sodium diet had significantly smaller endolymphatic sac lateral intercellular spaces than those animals fed only a high-sodium diet (analysis of variance with Scheffe's multiple comparison test, P less than 0.001). Animals fed a high-sodium diet had significantly larger endolymphatic sac lateral intercellular spaces than those animals fed a control diet only (analysis of variance with Scheffe's multiple comparison test, P less than 0.001). Results of this study suggest that dietary sodium affects endolymphatic sac fluid dynamics and that other food factors may regulate sodium metabolism, and therefore endolymphatic sac function.

We investigated the effects of furosemide, a loop diuretic, and canrenoate, an aldosterone antagonist, on the endocochlear potential (EP) and the endolymphatic sac potential (ESP) in the guinea pig. Furosemide produced no significant change in the ESP at a dose of 100 mg/kg after an intravenous infusion for 20 min. However, this dose decreased the EP to a negative level. Canrenoate produced no significant change in the EP at an intravenous dose of 300 mg/kg for 20 min, but it did decrease the ESP. The differences in the EP and ESP in the response to the diuretics indicate a dissimilarity of the origin of both d.c. potentials in the endolymphatic space.

The well-known toxicity pattern of aminoglycoside antibiotics has been used with the aim to produce waste products in the inner ear. The endolymphatic sac was studied after daily intraperitoneal injection of 75 mg/kg kanamycin for 10 and 20 days to see whether or not the endolymphatic sac showed signs of increased phagocytic activity. The epithelium of the endolymphatic sac was reduced in height, the cells were swollen and extremely thin. The dark epithelial cells were outnumbering the light cells. The subepithelial tissue showed an edematous swelling containing few blood vessels, collagen fibrils and fibroblasts with an empty appearance. There were few wide lateral intercellular spaces and there were intraluminal free floating cells. There were, in all, no clear morphological signs of increased phagocytic activity in the epithelium of the endolymphatic sac.