Anionic sites on Reissner's membrane, stria vascularis, and spiral prominence.

We demonstrated anionic sites on the lateral wall of cochlear duct and Reissner's membrane (RM) of ICR mice by Lowicryl K4M resin post-embedding and poly-L-lysine-colloidal gold conjugate (PL-CG) as a polycationic probe. The basement membrane and endolymphatic cell surface of RM were labeled with PL-CG pH 2.5 and pH 1.0. However, the perilymphatic cell surface was not labeled. PL-CG pH 2.5 and pH 1.0 strongly labeled the endolymphatic surface of the spiral prominence epithelium (SP), whereas the endolymphatic surface of the marginal cell (MC) in the stria vascularis was not labeled. Pre-digestion with several glycosidases eliminated PL-CG labeling. Our result suggests that an anionic charge located on the basement membrane of RM is largely due to the presence of heparan sulfate, chondroitin sulfate, and hyaluronic acid. An anionic charge on the endolymphatic cell surface of RM was mainly dependent on the presence of heparan sulfate. An anionic charge on the SP epithelium was caused to a substantial degree by chondroitin sulfate. We obtained histochemical evidence that the glycoconjugate content of the MC surface was quite different from that of the endolymphatic cell surface of RM and SP. We also identified RM-MC and SP-MC junctions at the ends of the stria vascularis between the marginal cells and the other endolymphatic epithelial cells of the cochlear duct.

Anionic sites in blood capillaries of the mouse cochlear duct.

The blood-cochlear barrier, which consists of the molecular size and ionic charge barriers, is known to play an important role in production and absorption of inner ear fluids. In this study, we employed poly-L-lysine colloidal gold conjugates (PL-CG) in combination with Lowicryl K4M resin to demonstrate anionic sites in blood capillaries of the cochlear duct. Male ICR mice weighing 30-40 g with a positive Preyer's reflex were used. The basement membranes of blood capillaries of the stria vascularis and the spiral ligament were successfully labeled with PL-CG pH 2.5. The luminal surface of capillaries in the stria vascularis and the spiral ligament intensely reacted with PL-CG pH 2.5. However, PL-CG pH 1.0 stained only the basement membrane of the spiral ligament. Predigestion with several glycosidases nearly eliminated PL-CG labeling. Anionic charge located on the luminal surface of the endothelial cell was mainly caused by the presence of sialic acid. On the contrary, anionic charge of the basement membrane was caused in a substantial degree by chondroitin and heparan sulfate-rich glycosaminoglycans. We obtained histochemical evidence that blood capillaries of the stria vascularis differ from those of the spiral ligament.

Effects of contrast media on the stria vascularis.

The effects of contrast media on the stria vascularis were electron microscopically studied in mice after intravenous injection of amidotrizoate, iopamidol or ioxaglic acid at 10 or 20 ml/kg body weight. In furosemide-load groups, contrast media were injected via the tail vein at 10 ml/kg with preadministration of furosemide at 20 mg/kg body weight. In both groups, the stria vascularis was prepared for study 10 min after the injection of contrast media. No morphological changes could be found after administration of 10 ml/kg injection of any of the three contrast media. However, administration of 20 ml/kg of amidotrizoate caused a high degree of intercellular dilatation between marginal and intermediate cells. Iopamidol caused a slight degree of dilatation, and ioxaglic acid caused no change. In the furosemide-load groups, amidotrizoate induced severer effects than iopamidol while ioxaglic acid induced no effect. From these results it was concluded that contrast media affect the stria vascularis parallel with osmolality, although some ionic effect of amidotrizoate should be considered when it is used in combination with furosemide.

Histopathology of eighth cranial nerve of mass stranded dolphins at Goto Islands, Japan.

We examined four dolphins (Grampus griseus) of 582 mass-stranded. Almost no contents were found in the alimentary canal. Nasitrema gondo and Crassicauda grampicola were found in the tympanic cavity. Severe degeneration of the eighth cranial nerve was observed microscopically in all animals and an egg of Nasitrema was found in a tissue crevice of the nerve. We propose that the nerves were damaged directly by Nasitrema.