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Scanning electron microscopy of the connective tissue along the lateral wall of the mouse cochlear duct with special reference to the external sulcus cells.

The external sulcus cells in the lateral wall of the cochlear duct insert cell cords into the connective tissue. To examine the shape, arrangement, and distribution of the sulcus cell cords in the cochlear duct in adult mice, the external sulcus cells were removed by chemical maceration methods, and the connective tissue exposed. The holes for the insertion of the sulcus cell cords along the entire course of the cochlear duct could thus be observed by scanning electron microscopy. A zone perforated with the holes was seen in the lateral region of the area between the stria vascularis and the lateral edge of the basilar membrane, this zone extending from the basal end of the hook to the helicotrema. In the hook and basal half turn, the holes possessed large fusiform openings and branched in the connective tissue toward the outer bony wall. In the apical turn, the holes displayed small round openings and were shallow, having a small number of branchings. The width of the perforated zone, the population density of the openings of the holes and the area proportion of the openings per unit surface area in the lateral wall decreased from the base to the apex of the cochlear duct. The regional differences in the organization of the sulcus cell cords suggest that the external sulcus cells function in close relation to regional auditory functions in the cochlear duct.

Animals

Scanning electron microscopy of the auditory teeth along the mouse cochlear duct.

The auditory teeth in the spiral limbus of the cochlear duct are located under the limbal portion of the tectorial membrane and separated by furrows lodging the interdental cells. In this study, the shape, arrangement and distribution of the auditory teeth in the cochlear duct of adult mice were examined by scanning electron microscopy after removing the tectorial membrane and the interdental cells with chemical maceration methods. The auditory teeth appeared on the top face of the spiral limbus between the edge of the vestibular lip and the Reissner membrane. The teeth on the vestibular lip side possessed elongated upper plates and formed a continuous row resembling the keyboard of a piano; the teeth were separated by radially oriented parallel slits. The elongated teeth decreased in length from the base to the apex of the cochlear duct. The teeth on the Reissner membrane side showed star-shaped upper plates separated by slits and gaps. The population density of the star-shaped teeth decreased from the base to the apex, widening the gaps to hold the interdental cells. The upper plates of the teeth occupied about 75% of an extent of the tooth zone in the hook and first basal half turn, and about 55% in the apical turn. The regional differences of the auditory teeth are considered to be closely related to local functions of the tectorial membrane and the interdental cells.

Animals

Effects of endolymphatic sac obliteration on the cochlear duct glycocalyx.

The ultrastructural effects of surgical obliteration of the endolymphatic duct and sac on the cochlear duct glycocalyx were investigated using osmium tetroxide-potassium rutheniumcyanide postfixation. After 2 and 3 months a less prominent glycocalyx contrast staining was observed, together with disappearance of interconnecting material between the stereocilia of the inner and outer hair cells. Possible factors involved in this phenomenon are discussed.

Animals

Utricle, saccule, and cochlear duct in relation to stapedotomy. A histologic human temporal bone study.

This study was performed to determine the area in which and the circumstances under which stapedotomy can be relatively safely performed. Measurements were made from central areas of the medial surface of the stapedial footplate to the utricle, the saccule, and the cochlear duct in 10 normal and 11 otosclerotic temporal bones. The mean distances to the utricle ranged from 1.9 to 2.4 mm, and those to the saccule from 1.7 to 2.1 mm. The minimal distance to the utricle was measured from the posterior (0.58 mm) and superior (0.62 mm) borders of the stapedial footplate. The minimal distances to the saccule were from the anterior (0.76, 0.86, and 1.00 mm) border of the stapedial footplate. All other measurements were of more than 1 mm. The shortest distance between the cochlear duct and the inferior border of the footplate was 0.2 mm. Statistical analysis has shown no significant differences for the mean values obtained in normal and otosclerotic temporal bones. Fathoming of the vestibule below the central and inferior thirds of the footplate surface has shown that there is no likely danger to the vestibular end organs or cochlear duct if manipulations are carried out no deeper than 1 mm below the surface. The safest place for a stapedotomy opening is in the central and inferior-central thirds of the footplate. A stapedotomy piston of 0.4 mm in diameter can be introduced relatively safely to a depth of 0.5 mm in the vestibule over the entire surface of the stapedial footplate.

Aged

Ionic activities of the inner ear fluid and ionic permeabilities of the cochlear duct in endolymphatic hydrops of the guinea pig.

Ionic activities (K+, Na+, and Cl-) of the perilymph and endolymph of the basal turn were measured using ion-selective microelectrodes in experimentally induced endolymphatic hydrops of the guinea pig. Three months following the obstruction of the endolymphatic duct and sac, the endocochlear potential (EP) of hydroptic ears was measured at 59.7 +/- 9.6 mV (N = 12) which was significantly lower than the EP of the contralateral control ears (84.4 +/- 2.8 mV, N = 12). A paired t-test (P greater than 0.05) showed no significant differences of ion concentrations of the inner ear fluid between the hydroptic and contralateral ears. Ion permeabilities of the cochlear duct following anoxia were calculated according to the Nernst-Planck equation. Comparing hydroptic and normal ears following anoxia, a statistically significant decrease was observed in the permeability coefficients for K+. Similarly, K+ conductance was significantly lower in the hydroptic ears than in the normal ears. Total conductance of the cochlear duct, defined as the sum of each ion conductance, was 0.560 siemens in the normal ears and 0.217 siemens in the hydroptic ears. On the basis of the Goldman-Hodgkin-Katz equation, preexisting negative EP in the normal state was calculated to be -24.5 mV in normal ears and -21.4 mV in hydroptic ears. Therefore, the positive component of the EP was 108.9 mV in normal ears and 81.1 mV in hydroptic ears. These findings suggest that the pathophysiology of hydrops involves changes in K+ permeability and the inhibition of the electrogenic transport processes.

Animals

Activation of muscarinic cholinergic receptors stimulates inositol phosphates synthesis in the developing avian cochlear duct.

We previously reported that the inositol phosphates (IPs) synthesis is induced by muscarinic agonists in the rat cochlea and that this stimulation is maximal at postnatal day 12. This peak response is concomitant with the onset of the efferent synaptogenesis at the outer hair cell level. Whether the correlation between this neuronal plasticity and the enhanced IPs formation is unique to the rat or a general feature of the developing vertebrate cochlea is not known. To examine this question, we measured, in the presence of LiCl, the accumulation of (3H)-IPs induced by carbachol, in the developing chick cochlear duct during a period ranging from embryonic day (E) 8 to post-hatching day (P) 20. Carbachol (1 mM) causes a significant increase of IPs formation relative to basal values at all ages. This IPs accumulation is maximal at E8 (1854% of the basal level), then, rapidly decreases until P13 when it reaches a steady-state level of 294% of the basal level. Strikingly, this gradual decline in IPs formation is interrupted between E15 and E19, by a transient increase in IPs synthesis. This rise peaks at E16 with a stimulation value of 757% of the control level. This maximal stimulation is inhibited by atropine in a dose-dependent manner, as is the case at E9, suggesting the involvement of muscarinic receptors. Interestingly, the occurrence of the peak response is concomitant with the plastic events associated with the maturation of the efferent innervation of the cochlear duct. Thus, these results suggest that there may be a correlation between cochlear plasticity and enhanced IPs synthesis, which is not species-specific.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Quantitative immunogold localization of Na+,K(+)-ATPase alpha-subunit in the tympanic wall of rat cochlear duct.

Ultrastructural localization of Na+,K(+)-ATPase was quantitatively investigated in the tympanic wall of rat cochlear duct by use of the protein A-gold method, using an affinity-purified antibody against the alpha-subunit of rat kidney Na+,K(+)-ATPase. A moderate number of gold particles were found on the basolateral membrane of the interdental cells of the spiral limbus. A small number of gold particles were found on the basolateral surfaces of the border cells and Hensen's cells. On the inner and outer sensory hair cells, however, the plasma membranes were rarely labeled by gold particles. The general pattern of labeling densities in cochlear structures determined here and in a previous communication from our laboratory shows good correlation with the distribution of Na+,K(+)-ATPase activity as previously estimated biochemically, cytochemically, and autoradiographically.

Animals

Quantitative immunocytochemical localization of Na+,K+-ATPase alpha-subunit in the lateral wall of rat cochlear duct.

Ultrastructural localization of the alpha-subunit of Na+,K+-ATPase on the lateral wall of rat cochlear duct was investigated quantitatively by the protein A-gold method, using affinity-purified antibody against the alpha-subunit of rat kidney Na+,K+-ATPase. In the stria vascularis, gold particles were sparse over the endolymphatic luminal surface of the marginal cells but were numerous over the basolateral membrane. The labeling density of the basolateral membrane was almost equal to that of the same domain of the distal tubule cells of kidney. The intermediate cells were studded with a large number of gold particles on the plasma membrane domain facing the basolateral domain of the marginal cells. On the luminal surfaces of the other epithelial cells, including those of Reissner's membrane, no significant amount of gold particles was found. Many gold particles were localized on all the plasma membranes of the spiral prominence stromal cells and on the intracellular membrane domain of the external sulcus cells.

Animals

[Light and electron microscopic studies on the development of the spiral prominence of the cochlear duct of the fetal guinea pig].

The anlage of the spiral prominence can be seen on the 37th day of development as a small protrusion of the epithelium towards the lumen of the cochlear duct. During the further progress, the spiral prominence more distinctly protrudes by augmentation of the vascularized connective tissue. In the epithelial cells pinocytotic vesicles near the plasmalemma are seen earliest lateral and basal on the 37th day, apical on the 39th day. The epithelial cells send basal cytoplasmic extensions towards the connective tissue. Starting on the 44th day, small invaginations of connective tissue extend into the epithelium, remaining separated from the epithelial cells by the basal lamina. Until the 48th day, the monostratified epithelium remains columnar, thereafter it changes to cuboidal or flat. Towards the end of the development, the invaginations of the connective tissue nearly reach the surface of the epithelium, being separated from the endolymph by a small epithelial area.

Animals

Single-cell layer membrane covering the degenerated cochlear duct after perilymphatic perfusion of streptomycin.

Ultrastructural changes of the extrasensory epithelium in the scala media of guinea pig cochleae were observed from 3 to 137 days after perilymphatic perfusion with 20% streptomycin. The degeneration started in the organ of Corti, progressed to the interdental cells and roots of the outer sulcus cells and finally involved other epithelial cells such as Claudius and the inner and outer sulcus cells. In the final stage, except for the stria vascularis and Reissner's membrane, all epithelial cells which lined the cochlear duct were replaced by a single-cell layer membrane which originated medianly from the epithelial cells of Reissner's membrane and laterally from the superficial outer sulcus cells.

Animals

[Light and electron microscopic studies of the greater epithelial ridge and its relationship to the developing tectorial membrane in the cochlear duct of the guinea pig (author's transl)].

In early stages of fetal development (36th day, 3rd turn) the thickening of the epithelium at the basal side of the cochlear duct forms two ridges. Later in fetal development the laterally situated lesser epithelial ridge forms the major part of the organ of Corti, whereas the medially situated greater epithelial ridge contributes only a small part to this organ. The medial part of the greater ridge consists of the columnar inner supporting cells, which bear a border of closely packed microvilli at their upper surface. Up to the time of the opening of the internal spiral sulcus in the 48th day of fetal development, there is a close spacial relationship between microvilli and filaments of the tectorial membrane. We conclude that the inner supporting cells contribute to the formation of the tectorial membrane. However, thus far we cannot entirely exclude a different possibility, that the inner supporting cells absorb material of the tectorial membrane. During the opening of the sulcus spiralis internus the inner supporting cells become considerably smaller, some of them undergo complete destruction by cytolysis, with pyknosis and karyorrhexis.

Animals

Single mitotic center for rodent cochlear duct.

The pattern of terminal mitosis in mouse otocyst observed by Ruben led him to postulate the existence of a growth zone at the junction of saccular and cochlear primordia. With the use of colchicine, an antimitotic drug, a localized zone of mitotic activity has been demonstrated at this predicted site.

Animals

The limbus spiralis and its relationship to the developing tectorial membrane in the cochlear duct of the Guinea pig fetus.

The development of the interdental cells of the limbus spiralis and of the inner spiral sulcus cells as well as the formation of the mesenchymal teeth of Huschke are described during fetal life up to the day of birth in the guinea pig. Additionally, the changes of the developing tectorial membrane are studied. The ultrastructural observations allow the conclusion that during fetal development at least a considerable part of the material of the tectorial membrane is secreted by the interdental cells of the limbus spiralis.

Animals