Preservation and visualization of actin-containing filaments in the apical zone of cochlear sensory cells

The fine filamentous structure in the apical zone of cochlear sensory cells of the guinea pig was investigated under transmission electron microscopy (TEM) using various fixation methods. The true form of this structure, which is that of a dense core of sensory hairs and cuticular plates containing hair rootlets, has been hitherto unknown because of the selectively destructive effect of ordinary fixatives. We revealed the fine filamentous structure in great detail by fixing the specimens in tannic acid or by the modified glutaraldehyde--osmium fixation method, which can preserve action filaments during the procedures required to prepare the specimen for TEM. The filamentous structure gives the impression of a negatively stained image when prepared in this way. Filaments were packed regularly and tightly into dense cores which projected down deep into the cuticular plate as hair rootlets. Cross-striations were seen at intervals of 360 +/- 28 A along the packed filaments, a distance which is comparable to the periodicity of an actin paracrystal. The overall diameter of each filament was 83 A. In fact, the structure of dense cores and hair rootlets proved to be composed of actin paracrystals, probably containing some regulatory proteins. Cross-sectioned actin filaments in the paracrystal were arranged in an extremely regular hexagonal pattern. The characteristic filamentous texture in the cuticular plate was best seen in tissues that were pretreated with EDTA, and then fixed by tannic acid. It is probable that the greater part of the cuticular plate is composed of actin filaments and actin monomers, both containing Ca2+-dependent regulatory proteins. Utilizing the above ultrastructural findings, some functional models of this zone are proposed.