Electron Microscope Characterization of Carbohydrate Residues on the Body Wall of Xiphinema index

Abstract

The location of carbohydrate moieties on the outer cuticle of Xiphinema index was examined by electron microcopy using several different reagents: a) The periodic acid-thiosemicarbazide-silver proteinate reaction was used as a general stain for carbohydrates. In sectioned material it stained the canal system and deeper layers of the cuticle as well as the outer surface, b) Cationized ferritin at pH 2.5, which identifies carboxyl and sulfate groups, was used to identify sialic acid residues and also labelled parts of the canal system, c) Ferritin-goat anti rabbit IgG coupled to a DNP ligand was used to label either sialyl or galactosyl/N-acetyl-D-galactosaminyl residues, d) Ferritin hydrazide, a new reagent, was used for the ultrastructural localization of glyco-conjugates. Reagents c) (with appropriate antisera) and d) were applied only to the outer surfaces of the cuticle; they showed that sialic acid residues were concentrated mainly on the outer body wall of the head, the lips, oral opening, amphid apertures, and outer surface of protruded odontostyles. Ferritin distribution was not altered by pretreatment with neurantinidase. Galactose oxidase treatments revealed galactose/N-acetyl-D-galactosamine residues along the entire body wall. These results confirmed earlier findings obtained by fluorescence microscopy. Key words: outer cuticle surface, cuticular canal system, sialic acid, galactose, electron microscopy, cryoultramicrotomy, cationized ferritin, ferritin hydrazide.