Ultrastructural properties of ciliary zonule microfibrils

Conventional electron microscopy and rotary shadowing techniques have provided conflicting interpretations of microfibril ultrastructure. To address this issue, we have used quick-freeze deep-etch (QFDE) microscopy to obtain 3-dimensional surface views of microfibrils that have not been fixed, dehydrated, or stained with heavy metals. By this approach, microfibrils appear as tightly packed rows of bead-like subunits that do not display the interbead filamentous links seen by other methods. At regular 50-nm intervals along the microfibril length, a larger bead is often recognized which tends to be aligned with those from adjacent microfibrils when the microfibrils are in bundles. This evidence of organized lateral associations of microfibrils is supported by the observation of small filaments that span between the adjacent microfibrils. When QFDE microscopy was used to examine microfibrils exposed to sonication, partially dissociated microfibrils with the more typical "beads on a string" appearance were observed. Beads are also seen alone, as monomers, often with an array of small thread-like filaments extending from the bead in a "crab-like" manner. Our results suggest that the beads on a string appearance of sonicated microfibrils may result from a partial loss of protein components from the interbead domains, thus leading to exposure of a filamentous substructure. It is possible, therefore, that this phenomenon might also contribute to the beads on a string appearance of microfibrils seen using other electron microscopy techniques.