Galabiosylceramide is present in human cerebrospinal fluid

• Published in: Biochemical and biophysical research communications Vol. 536; pp. 73 - 79
• Main Authors: Akiyama, Hisako, Ide, Mitsuko, Yamaji, Toshiyuki, Mizutani, Yasuaki, Niimi, Yoshiki, Mutoh, Tatsuro, Kamiguchi, Hiroyuki, Hirabayashi, Yoshio
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15.01.2021
• Subjects: Biosynthetic Pathways; ceramides; Cerebrospinal fluid; constitutional isomers; Fabry disease; Galabiosylceramide; Galactosyltransferases - metabolism; Gangliosides - biosynthesis; Gangliosides - cerebrospinal fluid; Gangliosides - chemistry; Glycolipid; Glycosphingolipids - isolation & purification; Glycosyltransferases - metabolism; HeLa Cells; Humans; hydrocephalus; Hydrocephalus - cerebrospinal fluid; hydrophilic interaction chromatography; Hydrophilic interaction chromatography–electrospray ionization-tandem mass spectrometry; ionization; kidneys; Lactosylceramide; liver; lysosomes; Shiga toxin; tandem mass spectrometry; urine; Lactosylceramide; HFA; NNC; Shiga toxin; NFA; CNS; MRM; Cerebrospinal fluid; iNPH; GalCer; UGCG; FBS; Gb2; Stx; CSF; Gb4; Gb3; STEC; A4GALT; LSD; HILIC; Hydrophilic interaction chromatography–electrospray ionization-tandem mass spectrometry; HPTLC; LacCer (d18:1-C12:0); GSL; Galabiosylceramide; AD; HUS; Glycolipid; GlcCer (d18:1-C12:0); CGT; HexCer; LacCer; LC; ESI-MS/MS; GlcCer; GLA
• ISSN: 0006-291X; 1090-2104; 1090-2104
• DOI: 10.1016/j.bbrc.2020.12.051

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. •Ceramide dihexoside was isolated from human CSF.•Ceramide dihexoside isolated from human CSF reacted with Shiga toxin.•LC/MS revealed the presence of LacCer and Gb2 in the isolated ceramide dihexoside.•The presence of Gb2 in human CSF was demonstrated for the first time.