Demonstration of Intermediate Filament Proteins in the Guinea Pig Vestibular Labyrinth by a New, High-resolution Cryotechnique

Abstract

In order to establish the more precise localization of IFPs in the vestibular labyrinth we have developed an immunohis-tochemical method using semithin cryosections from guinea pig inner ear. The vestibular end organs were fixed by intralabyrinthine perfusion with formaldehyde and glutaraldehyde. Microdissection was performed, followed by decalcifi-cation in EDTA. After specimen embedding in gelatin, semithin cryosections (1 μm) were prepared. Prior to the immunohistochemical staining, a new antigen-unmdsking treatment was performed. Monoclonal antibodies for cytoker-atins (Cks), vimentin, neurofilament protein (NF), and glial fibrillary acidic protein (GFAP) were used and visualized with an indirect ABC method. In the guinea pig vestibular labyrinth, Cks8, 18 and 19 were present. Virnentin and NF were stained positively, GFAP negatively. These results are in accordance with our previous results in the human vestibular labyrinth except for Ck7. The high-resolution cryotechnique in combination with a new antigen-unmasking method may yield more, and more detailed information about the localization of IFPs in the vestibular region.

Key Words:

• cytoskeleton
• antigen-unmasking method
• thinner cryosections
• immunohistochemistry