Arrangement of subunits in microribbons from Giardia.

Abstract

Ultrasound has been used to disperse the cytoplasm of Giardia muris and Giardia duodenalis trophozoites, releasing disk cytoskeletons for negative staining and study by electron microscopy. Sonication also breaks down the corss-bridges uniting microribbons in disks. Individual ribbons and small bundles of these structures, are found in these preparations and have been imaged both from their edges and in flat face view. The outer layers of ribbons are 2 sheets of regularly arranged globular subunits, held apart by a fibrous inner core. The axial repeat of the microribbon is 15 nm, which is also the distance separating cross-bridge sites along ribbons. Pronounced striping at this interval is a feature of ribbon faces where they are joined in bundles. Subunits in the outer layer are arranged in vertical protofilaments that are set orthogonally to the long axis of the ribbon. Protofilaments bind tannic acid and are seen clearly in sectioned ribbons. Three protofilaments fit into the 15-nm longitudinal spacing. Optical diffraction patterns from ribbon images are dominated by orders of the 15-nm periodicity, including the third-order reflexions expected from protofilaments spacings. Fourth-order reflexions indicate that the ribbon core may also be structured. Ribbon face images give rise to a strong 4-nm layer line, corresponding to the vertical spacing of subunits in protofilaments. Neighbouring protofilaments are staggered by about 0.67 nm. The lattices found in ribbons are consistent with studies of cytoskeleton composition.