Galabiosylceramide is present in human cerebrospinal fluid

Cerebrospinal fluid (CSF) contains glycosphingolipids, including lactosylceramide (LacCer, Galβ(1,4)Glcβ-ceramide). LacCer and its structural isomer, galabiosylceramide (Gb₂, Galα(1,4)Galβ-ceramide), are classified as ceramide dihexosides (CDH). Gb₂ is degraded by α-galactosidase A (GLA) in lysosomes, and genetic GLA deficiency causes Fabry disease, an X-linked lysosomal storage disorder. In patients with Fabry disease, Gb₂ accumulates in organs throughout the body. While Gb₂ has been reported to be in the liver, kidney, and urine of healthy individuals, its presence in CSF has not been reported, either in patients with Fabry disease or healthy controls. Here, we isolated CDH fractions from CSF of patients with idiopathic normal pressure hydrocephalus. Purified CDH fractions showed positive reaction with Shiga toxin, which specifically binds to the Galα(1,4)Galβ structure. The isolated CDH fractions were analyzed by hydrophilic interaction chromatography (HILIC)-electrospray ionization tandem mass spectrometry (ESI-MS/MS). HILIC-ESI-MS/MS separated LacCer and Gb₂ and revealed the presence of Gb₂ and LacCer in the fractions. We also found Gb₂ in CSF from neurologically normal control subjects. This is the first report to show Gb₂ exists in human CSF.