Summary
The presence of cell surface carbohydrates was investigated in intact, non-fixed outer hair cells (OHCs) of guinea pigs using fluorescein isothiocyanate (FITC) and rhodamine (TRITC) lectins. By means of wheat germ agglutinin (WGA) N-acetyl-d-glucosamine was shown in the entire OHC membrane, including the stereocilia. Binding of WGA in OHCs to neuraminic acid was excluded by preincubation with neuraminidase. Moreover, FITC-Limulus polyphemus, a specific lectin for neuraminic acid, showed no fluorescence on OHCs. Neutral saccharides, like α-d-mannose and/or α-d-glucose, were mainly observed at the cuticular plate and at the basal cell pole with FITC-concanavalin A. A weaker fluorescence was seen at the lateral cell wall. Two branched oligosaccharides, composed of β-galactose, N-acetyl-d-glucosamine and mannose, were demonstrated by TRITC-Phaseolus vulgaris (PHA-E/L) in the entire OHC membrane. A spot-like binding of soybean agglutinin to N-acetyl-D-galactosamine could be demonstrated in the region of the cuticular plate. However, using Helix pomatia, the subtype N-acetyl-α-d-galactosamine was not detectable. Moreover, there was no binding of Ulex europaeus or of Arachis hypogea (PNA) to OHCs, suggesting the absence of considerable amounts of L-fucose and of galactose-β-3-N-acetylgalactosamine. The results indicate that N-acetyl-d-glucosamine, α-d-mannose and α-d-glucose can be considered the major components of the OHC glycocalix. We suggest that they have a function as an anchoring structure in interstereociliary links as well as in hair bundle connections to the tectorial membrane. Branched oligosaccharides identified by PHA-E/L could indicate the presence of cell-adhesion molecules in the OHC surface. In addition, a spot-like arrangement of N-acetyl-β-d-galactosamine near the cuticular plate may demonstrate junction-adhesion molecules, e.g. proteins of the zonula adherentes, in this region.
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Plinkert, P.K., Plinkert, B. & Zenner, H.P. Carbohydrates in the cell surface of hair cells from the guinea pig cochlea. Eur Arch Otorhinolaryngol 249, 67–73 (1992). https://doi.org/10.1007/BF00186449
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DOI: https://doi.org/10.1007/BF00186449