Effect of inner ear blood flow changes on the endolymphatic sac.

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.

Molecular mechanisms underlying inner ear patterning defects in kreisler mutants.

Prior studies have shown that kreisler mutants display early inner ear defects that are related to abnormal hindbrain development and signaling. These defects in kreisler mice have been linked to mutation of the kr/mafB gene. To investigate potential relevance of kr/mafB and abnormal hindbrain development in inner ear patterning, we analyzed the ear morphogenesis in kreisler mice using a paint-fill technique. We also examined the expression patterns of a battery of genes important for normal inner ear patterning and development. Our results indicate that the loss of dorsal otic structures such as the endolymphatic duct and sac is attributable to the downregulation of Gbx2, Dlx5 and Wnt2b in the dorsal region of the otocyst. In contrast, the expanded expression domain of Otx2 in the ventral otic region likely contributes to the cochlear phenotype seen in kreisler mutants. Sensory organ development is also markedly disrupted in kreisler mutants. This pattern of defects and gene expression changes is remarkably similar to that observed in Gbx2 mutants. Taken together, the data show an important role for hindbrain cues, and indirectly, kr/mafB, in guiding inner ear morphogenesis. The data also identify Gbx2, Dlx5, Wnt2b and Otx2 as key otic genes ultimately affected by perturbation of the kr/mafB-hindbrain pathway.

Erythrocyte removal and blood clearance in the endolymphatic sac. An experimental and TEM study.

The endolymphatic sac (ES) appears to serve as an immunologic/phagocytic defence organ. This may involve both specific and unspecific cell reactions for removal of both endogenous waste products and debris and foreign material. The present study was carried out in order to investigate whether the ES is involved in the removal of blood from the inner ear. The round window was exposed and the stapedial artery experimentally lesioned to cause bleeding. The ES was taken out after various time intervals and analyzed by TEM and LM, care being taken to maintain the structural integrity at all levels. This was done in order to see if the endolymphatic sac is involved in the removal of blood from the inner ear. The results indicate that the ES possesses hitherto unknown specific cellular properties for the disposal of blood and blood corpuscles. It is believed that this system may represent an important physiological function for regulation of homeostatic equilibrium around the inner ear sensory structures. The results also offer further support for the existence of a longitudinal "flow" of endolymph from the cochlea into the ES. Judging from the time it takes for erythrocytes to appear in the duct and sac, this flow would seem to take about 1-2 h in the mouse.

The endolymphatic duct and sac of the rat: a histological, ultrastructural, and immunocytochemical investigation.

A study of the ultrastructure, vascularization, and innervation of the endolymphatic duct and sac of the rat has been performed by means of light- and electron-microscopic and immunocytochemical methods. Two different types of epithelial cells have been identified: the ribosome-rich cell and the mitochondria-rich cell. These two cell types make up the epithelium of the complete endolymphatic duct and sac, although differences in their quantitative distribution exist. The morphology of the ribosome-rich cells varies between the different parts of the endolymphatic duct and sac; the morphology of the mitochondria-rich cells remains constant. According to the epithelial composition, vascularization, and structural organization of the lamina propria, both duct and sac are subdivided into three different parts. A graphic reconstruction of the vascular network supplying the endolymphatic duct and sac shows that the vascular pattern varies among the different parts. In addition, the capillaries of the duct are of the continuous types, whereas those supplying the sac are of the fenestrated type. Nerve fibers do not occur within the epithelium of the endolymphatic duct and sac. A few nerve fibers regularly occur in the subepithelial compartment close to the blood vessels; these fibers have been demonstrated in whole-mount preparations by the application of the neuronal marker protein gene product 9.5. Single beaded fibers immunoreactive to substance P and calcitonin-gene related peptide are observed within the same compartment. Dopamine-beta-hydroxylase-immunoreactive axons are restricted to the walls of arterioles. Morphological differences between the different portions of the endolymphatic duct and sac are discussed with regard to possible roles in fluid absorption and immunocompetence.

Fine structure of the endolymphatic duct in the rat. A scanning and transmission electron microscopy study.

To investigate the surface morphology of the endolymphatic duct epithelium, 8 rats were vascularly perfused with glutaraldehyde in a buffered and oxygenated blood substitute. Optimal preservation of the epithelium for scanning electron microscopy was attained by coating of the specimens with OsO4 and thiocarbohydrazide followed by a continuous dehydration procedure. Three types of cells were identified with the scanning electron microscope: A polygonal and oblong epithelial cell was observed in the largest number throughout the duct, whereas in the juxta-saccular half of the duct two additional types of epithelial cells were observed. The scanning electron microscopical observations are compared and discussed with reference to transmission electron microscopical observations of the endolymphatic duct.

Ultrastructure and barrier properties of the endolymphatic duct in the guinea pig.

The ultrastructure and barrier properties of the endolymphatic duct (ED) were examined by light and electron microscopy. ED epithelial cells were classified into two types: type I and type II duct cells. The type I duct cells were cuboidal or low columnar and were characterized by a convex apical surface and a few basal processes. The type II duct cells were squamous and were characterized by a flat apical and basal membrane, many small vesicles and a number of small pits along the basal membranes. After electrophoretic horseradish peroxidase (HRP) injection into the ED lumen, no HRP uptake into ED epithelial cells of either type was observed. There was no reaction product either in the lateral intercellular spaces beyond the tight junctions between ED epithelial cells or in ED subepithelial tissues. The ED epithelial cells were considered to play no active role in apical macromolecular absorption and to be impermeable to intraluminal macromolecules.

Contribution of cell culture to the study of the physiology of the stria vascularis and Reissner's membrane.

Cell culture is a well-established tool in cell biology which may overcome the limitations of the experimental methods currently used to investigate the physiology of labyrinthine fluids. Using explant and dissociated cell culture technique, sheets of oriented cells derived from the stria vascularis and Reissner's membrane may be obtained, which allows new experimental approaches such as the patch-clamp technique. Other experimental approaches are considered which might improve our understanding of inner ear physiology.

Microorganism transport in the human endolymphatic duct.

There are indications that endolymphatic sac (ES) may be an immunologically active part of the inner ear. So far, no microorganisms or foreign substances have been localized in this area under 'normal' conditions. Only a limited number of human specimens, including the entire endolymphatic duct (ED) and ES, have been collected and analyzed from cadavers or surgical biopsy specimens. In this study, 6 human ED and ES collected from cadavers and at surgery were analyzed by light and electron microscopy. This was done in order to investigate if microorganisms may normally be drained at this route into the ES. Some microorganisms (Mycoplasma pneumoniae) were found in the lumen and subepithelial tissue of 1 human ED. These observations suggest that microorganisms may also be locally processed and disposed at the level of the ED. These results add further evidence as to the immunodefensive role of the human ES.

Differential immunohistochemical detection of cytokeratins and vimentin in the surgically removed human endolymphatic duct and sac.

Immunohistochemical detection of intermediate filament proteins and different subgroups of cytokeratins (Cks) was used to characterize the epithelium of the surgically removed adult human endolymphatic duct (ED) and sac (ES). The epithelium of the ED and ES demonstrated immunostaining for Cks 7, 8, 14, 17, 18 and 19, a pattern typical of so-called "complex" or "mixed" epithelia. This is a remarkable finding, since this pattern differs strikingly from previously reported data on the adult human cochlea and vestibular labyrinth that demonstrated a Ck pattern typical of "simple" (or single-layered) epithelia. Furthermore, the epithelium of the ED and ES demonstrated co-expression of Cks and vimentin. The present data indicate that the epithelium of the ED and ES exhibits another type of epithelial differentiation and demonstrates a higher degree of complexity than the other epithelia in the adult human inner ear.

Morphology of the endolymphatic duct and sac in the Mongolian gerbil.

A light microscopical study of the endolymphatic duct and sac of the Mongolian gerbil is presented. This animal model was studied because of its tendency to develop a condition of body fluid imbalance which may represent a hazard to the inner ear fluid environment. Particular stress is laid on the combination of the lack of an extraosseous endolymphatic sac and the presence of elastic tissue in the subepithelial space of the sac. These findings highlight the role of this structure for the regulation of the pressure within the inner ear. Lastly, a secretory process is proposed by which a homogeneous precipitate produced in the lumen of the sac can regulate volume and pressure changes.