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Scanning electron microscopy of hair cells, stereocilia and cross-linkage systems in experimentally induced endolymphatic hydrops.

In this study scanning electron microscopy was used to examine the stereocilia and their cross-linkages in guinea pig cochleas 1, 2, 4 and 8 months after endolymphatic sac obliteration. Initial changes were restricted to the stereocilia of the outer hair cells and consisted of a disarrangement and bulging of the stereocilia with an interruption of their cross-linkage systems. Subsequently, the stereocilia became fused and atrophied. Cross-linkages of inner hair cells remained intact. Loss of both outer and inner hair cells started in the apex and progressed towards the base of the cochlea. These findings indicate that early changes in the micro-architecture of the stereocilia may have a mechanical origin, with pressure fluctuations in the scala media possibly playing an important role.

Animals↗

Cochlear permeability of neomycin and gentamicin: an immunohistochemical study.

Aminoglycoside antibiotics (AGAs) target specifically the cochleo-vestibular hair cells, but with varied ototoxicity. Differences in their penetration and clearance rates into the membranous labyrinth may play a role. This in turn may be related to a difference in the number of amine groups, the cationic nature, as well as the molecular weight and size of the AGA molecule. Immunohistochemical labeling techniques were used to study the pathways of gentamicin and neomycin from the perilymph into cochlear tissues and target cells. The more cochleotoxic AGA, neomycin, penetrated into cochlear tissues faster than the less cochleotoxic AGA, gentamicin.

Animals↗

Primary culture of vital marginal cells from cochlear explants of the stria vascularis.

Explants of the stria vascularis and spiral ligament were dissected from guinea pig cochleae and were successfully cultivated for several weeks. After 2 days, fibroblast-like cells of the spiral ligament covered the bottom of the cell culture dish around the explant. Marginal cells of the stria vascularis proliferated and grew on the luminal surface towards the border of the explant at a rate of 15 microns/day. At day 6 in culture the proliferating marginal cells reached the border of the explant and then advanced to the bottom of the cell-culture dish. There the marginal cells replaced fibroblast-like cells and built an epithelial hexagonal-shaped monolayer. Light microscopic and transmission electron microscopic investigations revealed that the cultured cells were viable and that typical morphological characteristics of marginal cells were preserved. Cultivation of these cells provides a unique model for studies of physiological properties of marginal cells of the stria vascularis.

Acridine Orange↗

Effects of low-sodium, high-potassium dietary intake on cochlear lateral wall Na+,K(+)-ATPase.

The effect of a low Na+, high K+ diet on Na+,K(+)-ATPase levels in cochlear lateral wall tissues was investigated in laboratory rats by using an enzyme-linked immunosorbent assay. The low Na+, high K+ diet induced high aldosterone plasma levels in the animals as well as changes in plasma cation levels. Animals that received a low Na+, high K+ diet demonstrated a statistically significant (97%) increase in Na+,K(+)-ATPase levels in the stria vascularis when compared to animals that received a control diet. This increase in strial Na+,K(+)-ATPase levels was blocked only 70% by administration of the aldosterone antagonist, spironolactone. Findings therefore indicate that strial Na+,K(+)-ATPase may be modulated by both aldosterone and Na+,K+ plasma levels. Na+,K(+)-ATPase levels in the spiral ligament were not affected by the experimental treatment. These findings suggest that spiral ligament Na+,K(+)-ATPase levels may be regulated by factors other than aldosterone and Na+,K+ plasma levels. This study provides further insight into the mechanisms of the beneficial effects of salt restriction and potassium loading in patients with Méniére's disease.

Aldosterone↗

Cytokeratin expression in the epithelia of the adult human cochlea.

Epithelia can be characterized by the specific expression pattern of their cytokeratin components. Therefore, we investigated the immunohistochemical expression of different cytokeratin subunits in frozen sections of chemically fixed, non-decalcified, adult human cochleas. The organ of Corti and the marginal cells of the stria vascularis showed reactivity for cytokeratin subunits 8, 18 and 19, whereas the other cochlear epithelia in addition expressed cytokeratin 7. The expression of cytokeratins 7, 8, 18 and 19 by the epithelia of the adult human cochlea is typical of "simple" epithelia. The deviant cytokeratin pattern of the organ of Corti and marginal cells of the stria vascularis may well reflect their differences in functional state and/or differentiation as compared to the other cochlear epithelia.

Adult↗

Ultrastructure of Reissner's membrane in the rabbit.

The ultrastructure of Reissner's membrane in the rabbit is described following vascular perfusion-fixation of live, anesthetized and artificially respirated healthy animals. A new and improved technique of fixation is employed that includes a pressure feedback controlled peristaltic pump and an oxygen-carrying fixative. In ultrathin sections capillaries were observed between the two cell layers comprising Reissner's membrane. The mesothelial cells facing the scala vestibuli were connected by junctional complexes and neither pores nor discontinuities were observed in the cell layer. In the epithelial cells a well-developed tubulocisternal endoplasmic reticulum (TER) was noted. Computerized three-dimensional reconstruction documented the continuity of this TER, from prominent disc-shaped subsurface cisterns lining the luminal cell membrane to smaller subsurface cisterns lining the abluminal and lateral cell membranes, forming a transcellular canalicular pathway. The possible function of the TER in Reissner's membrane is discussed with reference to endolymph/perilymph homeostasis.

Animals↗

Fine structure and permeability of capillaries in the stria vascularis and spiral ligament of the inner ear of the guinea pig.

The blood capillaries in the stria vascularis and the spiral ligaments of guinea pigs were studied by electron microscopy with freeze-fracture and thin section methods, including tracer experiments with horseradish peroxidase (HRP) and microperoxidase (MP). The endothelial cells of the capillaries of both tissues are connected by tight junctions, and contain about the same number of micropinocytotic vesicles. In cases of intravascular administration before fixation, both of the tracers stained the perivascular space and almost all endothelial vesicles in the stria vascularis. On the other hand, the perivascular space and many vesicles in the spinal ligament were unstained. The endothelial tight junctions in the stria vascularis prevented the penetration of HRP, but sometimes allowed the penetration of MP. Those of the spiral ligament were impermeable to both tracers. In cases of tracer administration after fixation, leakage spots of HRP from capillaries were sparsely located all over the stria vascularis. Transendothelial channels and isolated fenestrae formed by micropinocytotic vesicles were detected. It is concluded that the capillaries of the stria vascularis are similar to the muscle capillaries and to the capillaries of the elasmobranch brain, whereas those in the spiral ligament are similar to the brain capillaries of higher vertebrates.

Animals↗

A comparison of the reduction in the K+ activity of the scala media produced by furosemide and ouabain.

The endocochlear potential (EP) and the K+ activity in the scala media of the guinea pig were recorded by means of K(+)-sensitive, double-barrelled microelectrodes in the second turn of the cochlea. EP and K+ change were recorded during the perilymphatic perfusion of furosemide and ouabain. Furosemide produced less of a decrease than ouabain in the K+ activity relative to the EP decrease. These results suggest that furosemide reduces the EP via different mechanisms from the inhibition of the K+ secretion potential.

Action Potentials↗

Three-dimensional organization of a transcellular tubulocisternal endoplasmic reticulum in epithelial cells of Reissner's membrane in the guinea-pig.

The ultrastructure of the epithelial cells of Reissner's membrane (membrana vestibularis) in the guinea-pig is described following vascular perfusion with glutaraldehyde of live, anaesthetised and artificially respirated animals. Postfixation in a solution containing OsO4 and potassium ferricyanide revealed a well-developed tubulocisternal endoplasmic reticulum, not previously described, the continuity of which has been mapped by serial sectioning and reconstruction. Large disc-shaped subsurface cisternae lining the cell membrane, but separated from it by a space approximately 10 nm wide, are in continuity with the smooth endoplasmic reticulum, forming an elaborated transcellular canalicular pathway. This structure is compared to that found in solute-transporting epithelia, e.g., renal proximal tubule, gall bladder, small intestine and choroid plexus. The fixation method used in the present study is compared to other techniques used for preservation of Reissner's membrane. Each epithelial cell of Reissner's membrane is endowed with one kinocilium, one to four multivesicular bodies, and a number of intercalated bodies. The functional significance of the canalicular pathway is discussed.

Animals↗

The value of non-reconstructive multiplanar CT for the evaluation of the petrous bone.

The application of high resolution CT in the petrous bone has so far been restricted to the transverse and the coronal planes. Multiplanar reformatting in other planes suffers severely from loss of spatial resolution and from patient motion. This article describes how a multitude of classical otoradiological planes can be imaged by means of direct CT using proper patient positioning techniques; thus providing the advantage of visualizing the different anatomical details in their optimal cross-sectional planes. Patient positioning and examples of CT anatomy are presented in seven different planes: transverse, coronal, sagittal, semi-axial, semi-longitudinal, axio-petrosal and longitudinal. A survey is presented of which anatomical details can be visualized best in which planes.

Cochlear Duct↗

Maintenance of hydrostatic pressure gradients in the membranous labyrinth.

In normal guinea pig ears, hydrostatic perilymphatic pressure is equal to endolymphatic pressure. Alterations of perilymphatic pressure induced, for example, by laceration of the round window membrane are transmitted immediately to the endolymphatic compartment, probably via Reissner's membrane. In guinea pigs with experimental endolymphatic hydrops, however, pressure gradients between the endolymph and perilymph remained preserved after rupture of the round window membrane. This is considered as further evidence that after long-standing distention of Reissner's membrane the membranous labyrinth loses its ability to equalize endolymphatic and perilymphatic pressure.

Animals↗

[Morphologic studies of the toxicity of ethacrynic acid in the spiral prominence].

Ultrastructural changes in the guinea pig spiral prominence were studied at various times after a single intravenous injection of ethacrynic acid (40 mg/kg body-weight). Initial swelling of endolymph-facing epithelial cells was followed by dilatation of the intercellular spaces and marked shrinkage of the stroma cells surrounding the spiral prominence vessel. While the changes in the stria vascularis and the spiral prominence progressed at about the same pace, the regression to normal ultrastructure set in earlier in the spiral prominence.

Animals↗

X-ray microanalysis of fluid spaces in the frozen cochlea.

The cochlea is quick-frozen and then opened while under liquid nitrogen to expose the scalae of all turns in the mid-modiolar section. Still under liquid nitrogen, the cochlea is transferred to the cold stage of a pre-vacuum chamber especially constructed for attachment to the scanning electron microscope. Under partial vacuum, it is moved to the cold stage in the high vacuum chamber of a field-emission scanning electron microscope. Energy dispersive X-ray analysis demonstrates the relative levels of sodium, potassium, and chlorine in the fluid spaces of the cochlea.

Animals↗

Hereditary deafness in the dalmatian dog.

The temporal bones from 18 deaf Dalmatian dogs were examined with the light microscope, and the cochleo-saccular degeneration process studied. End organ degeneration is already advanced by the age of 4 weeks, whilst loss of neurons in the spiral ganglion was found only in adult animals. Both processes are first evident in the central portion of the cochlea, and increase in extent with advancing age, The implications of these findings for the management of non-conductive hearing loss in man are discussed.

Animals↗

Scanning ultrastructure of the stria vascularis.

In the eleven years since the introduction of the first commercially available scanning electron microscope, a number of otologic studies have made use of this new technique. However, the literature contains only one report of scanning ultrastructure of the endolymphatic surface of the stria vascularis. We have studied the stria vascularis with a technique allowing examination of the mid-modiolar cut surface as well as the endolymphatic surface.

Animals↗

Macromolecular transport in the spiral prominence.

Transport of macromolecules in the spiral prominence of the guinea pig was studied after perilymphatic and intravenous horseradish peroxidase (HRP) injection. HRP was taken up by the spiral prominence epithelium from the perilymphatic space and in a much higher degree from the endolymphatic space. A highly endocytic activity was shown by the stroma cells surrounding the spiral prominence vessel. HRP particles were stored in the cytoplasm within vesicles and vacuoles of different sizes and different contrasts. The mode of tracer storage in the spiral prominence epithelium and in the stroma cells from the morphological point of view suggests lysosomal digestion. HRP was transported from the perilymphatic space into the spiral prominence vessel lumen by means of vesicular transport through the vessel wall, while transport of the tracer from the vessel lumen into the perilymphatic space after intravenous injection was not observed.

Animals↗