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