Structural analysis and immunohistochemical localization of tow acidic glycosphingolipids from the porcine, parasitic nematode, Ascaris suum

Abstract

The acidic glycolipid fraction (AF) of the porcine, parasitic nematode, Ascaris suum, consisted of two subfractions. The major component AF II reacted with orcinol-sulfuric acid and molybdate, while the minor component AF I gave a positive reaction with azure-Ad, a cationic dye specific for sulfatides. Sugar constituent analysis, methanolysis, methylation analysis, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, liquid secondary-ion mass spectrometry, and gas-liquid chromatography/mass spectrometry specified AF II to be an unusual phosphoinositolglycosphingolipid (Gaα1-Ins-P-1ceramide) and the minor component AF I to be a 3-sulfogalactosoylcerebroside (HSO3-3Galβ1-1ceramide). The ceramide moiety of both components consisted of lignoceric (C24:0) and cerebronic (C24h:0) acids and mainly C17 iso-branched sphingosine. Immunohistochemical localization studies of the glycolipid-bound antigenic determinants with a polyclonal antiserum against AF II and an anti-sulfatide monoclonal antibody against AF I revealed the presence of the AF II-epitope in the intestine, whereas the AF I-epitope was found in the hypodermis, contractile zone of somatic muscle cells and the external musculature of the uterus. To our knowledge, this is the first report of the presence of a sulfatide in an invertebrate.