Auto-inflammatory challenge of the endolymphatic sac--Cochlear damage measured by distortion product oto-acoustic emissions.

CONCLUSION: Twenty-five rats were challenged by an immunologic attack of the endolymphatic sac. After 6 months, distortion product oto-acoustic emissions (DPOAE) revealed a dysfunction of the outer hair cells and immunological active cells were observed in the endolymphatic sac. This information could contribute to the understanding of Ménière's disease. OBJECTIVES: This study investigated if an autoimmune challenge of the endolymphatic sac could affect DPOAE output measurements in rats. Also, a potential autoimmune cell infiltration of the endolymphatic sac was investigated. METHODS: Eighteen Lewis rats were immunized with a crude endolymphatic sac extract in complete Freund's adjuvant. Seven control animals were injected with Freund's adjuvant in saline. Cochlear damage was estimated by DPOAE dynamics 3 weeks and 6 months after the immunization. Infiltrative cells in the endolymphatic sac were investigated with transmission electron microscopy. RESULTS: The hearing assessment 6 months after immunization revealed a reduction of the DPOAE, on the full range of frequencies (2-63 kHz) in an average of the mean, of 2 dB ± 1.1 in the immunized group compared to the controls (p < 0.05). The same test showed a 2.5 dB decrease from 2 to 5 kHz (p < 0.01). Immunological active cells were observed in the endolymphatic sac in most of the immunized rats.

Expression and localization of 11beta-hydroxysteroid dehydrogenase (11betaHSD) in the rat endolymphatic sac.

CONCLUSIONS: 11beta-Hydroxysteroid dehydrogenase type 2 (11bHSD-2) enables aldosterone to bind to mineralocorticoid receptors (MRs) selectively by converting cortisol (corticosterone) into inactive metabolites. Its expression in the endolymphatic sac (ES) suggests that aldosterone may selectively act on the ES through its binding to MRs by the action of 11betaHSD-2, and supports the notion that ES is an aldosterone target organ. We propose that 11betaHSD-2 is a dominant isoform of 11betaHSDs in the ES, and the ES (especially the intermediate portion of the ES) may be the main aldosterone target in the inner ear. OBJECTIVE: The purpose of this study was to examine 11bHSD isoform expression in the rat inner ear, mainly 11betaHSD-2 in the ES. MATERIALS AND METHODS: In the ES and whole cochlea, 11betaHSDs were examined by RT-PCR using highly specific ES RNA by laser capture microdissection. In addition, 11betaHSD-2 localization in the rat ES was determined by immunohistochemistry. RESULTS: RT-PCR demonstrated 11betaHSD-2 expressed in the rat ES. In addition, its localization was observed mainly in the intermediate portion and a faint immune positive signal was observed in other parts of the ES. In contrast, 11bHSD-1 was undetectable in the ES by RT-PCR. Both types of 11betaHSDs were expressed in rat cochlea.

Cloning and expression of vacuolar proton-pumping ATPase subunits in the follicular epithelium of the bullfrog endolymphatic sac.

In an investigation aimed at clarifying the mechanism of crystal dissolution of the calcium carbonate lattice in otoconia (the mineral particles embedded in the otolithic membrane) of the endolymphatic sac (ELS) of the bullfrog, cDNAs encoding the A- and E-subunits of bullfrog vacuolar proton-pumping ATPase (V-ATPase) were cloned and sequenced. The cDNA of the A-subunit consisted of an 11-bp 5'-untranslated region (UTR), a 1,854-bp open reading frame (ORF) encoding a protein comprising 617 amino acids with a calculated molecular mass of 68,168 Da, and a 248-bp 3'-UTR followed by a poly(A) tail. The cDNA of the E-subunit consisted of a 72-bp 5'-UTR, a 681-bp ORF encoding a protein of 226 amino acids with a calculated molecular mass of 26,020 Da, and a 799-bp 3'-UTR followed by a poly(A) tail. Western blot and immunofluorescence analyses using specific anti-peptide antisera against the V-ATPase A- and E-subunits revealed that these subunits were present in the ELS, urinary bladder, skin, testes, and kidneys. In the ELS, positive cells were scattered in the follicular epithelium which, as revealed by electron microscopy, corresponds to the location of mitochondria-rich cells. These findings suggest that V-ATPase, including the A- and E-subunits, exists in mitochondria-rich cells of the ELS, which might be involved in dissolution of the calcium carbonate crystals in the lumen of the ELS.

Intracellular and extracellular elemental composition of the endolymphatic sac studied by X-ray microanalysis.

X-ray microanalysis was performed along with light microscopy (LM) on rapidly frozen and cryo-sectioned endolymphatic sac tissues of adult guinea pigs, to determine the elemental composition of the different cell types in this tissue as well as the content of the sac lumen. The morphological preservation and spatial resolution of cryo-sectioned endolymphatic sac was found adequate for the identification of the different cell types of the sac in the transmission electron microscope. Further cell type identification was performed by comparing scanning transmission electron microscopy images with LM images on adjacent serial sections. X-ray microanalysis demonstrated differences between epithelial and sub-epithelial cells in the intracellular concentrations of sodium, chlorine, potassium, calcium, phosphorus and sulphur. Measurements performed in the lumen of the endolymphatic sac showed elevated sodium and decreased potassium levels as compared with the known levels of these elements in cochlear or vestibular endolymph. High phosphorus and sulphur levels were also found in the endolymph of the sac. Other morphological and analytical findings on the luminal content point out that otoconial destruction and cleaning of the endolymph from the cell debris and other products such as lipids and proteins take place in the endolymphatic sac. Our results suggest that the endolymphatic sac participates in fluid absorption (osmoregulation), ion transport and otoconial destruction. The data support the longitudinal flow theory of the endolymph.

Effects of high intensity pure tone stimulation on the endolymphatic sac. Correlations between cochlear morphology and endolymphatic sac response.

A systematic study of the effects of acoustic overstimulation on the endolymphatic sac (ES) in the guinea pig was performed. The ES was studied with light and transmission electron microscopy after exposure of the animals to a 3.85 kHz pure tone of 108 dB SPL or 120 dB SPL for 22.5 min (sound energy 9.4 and 150 Pa2 X h, respectively). The damage pattern in the organ of Corti was studied after various post-exposure times with SEM and correlated with the morphological characteristics of the ES in the same ear. This was made possible by using a modified technique for histological processing. In ears with induced structural abnormalities to the organ of Corti, the ES displayed few morphological changes without obvious signs of accumulation of cell debris within the lumen. Initially an increase in the amount of freely floating cells was found which persisted for at least 24 h. The role of the ES for disposal and digestion of locally produced degeneration products within the cochlea after acoustically generated structural damage is discussed.