Stage-specific expression of fuco-neolacto- (Lewis X) and ganglio-series neutral glycosphingolipids during brain development: characterization of Lewis X and related glycosphingolipids in bovine,human and rat brain

We have purified and characterized a bovine brain pentaglycosylceramide as Lewis X and identified it in human and rat brain using anti-Lewis X (anti-SSEA 1) monoclonal antibody. Neutral glycosphingolipid expression in developing rat brain has been examined by digoxigenin immunostaining and TLC-immunostaining using anti-SSEA 1 and anti-GgOse₄Cer (GA1) monoclonal antibodies. Five transient Lewis X-series bands were identified in brain at embryonic day 15 that disappear by postnatal day 5 (one disappears at embryonic day 18). Gangliotetraosylceramide (GA1) first appears at embryonic day 21 and increases in concentration with age until postnatal day 21. In addition, we have purified another minor brain neutral glycosphingolipid and tentatively identified it as a Lewis X-series glycolipid by gas chromatography-mass spectrometry analysis followed by TLC-immunostaining with anti-SSEA 1 antibody.Abbreviations Cer Ceramide, GlcCer, Glc1-1Cer - LacCer Gal1-4GlcCer - CTH Gal1-4LacCer - nLcOse₄Cer Gal1-4GlcNAc1-3LacCer - nLcOse₅Cer Gal1-3nLcOse₄Cer - GgOse₄Cer Gal1-3GalNAc1-4LacCer - DPA diphenylamine-aniline-phosphoric acid - SSEA stage-specific embryonic antigen - NGSL neutral glycosphingolipid - TLC thin-layer tomography - HPTLC high performance thin-layer chromatography - GA1 gangliotetraosyleramide - SAT-2 sialytransferase-2 - GalNAcT-1 galactosaminyltransferase-1 - DIG-IS digoxigenin-immunostaining - PMAAS partially methylated alditol acetates - DCE dichloroethane - TLC-IS TLC-immunostaining - (Le^x) Lewis X - NK murine natural killer