Chemical and Immunological Characterization of Galactosyl‐β1‐3‐Globoside in Bovine, Human, and Rat Brain

Abstract

Our studies of bovine brain neutral glycosphingolipids (Ngsls) have revealed the presence of several short‐chain (containing ‐CHO 1–4) and previously uncharacterized long‐chain (−CHO > 4–5) Ngsls. We reported the structural characterization of brain GgOse4Cer (GA1) and have now purified another brain Ngsl to homogeneity. The purified Ngsl migrated close to standard GgOse4Cer and nLcOse5Cer on a TLC plate employing two different solvent systems. The carbohydrate molar composition indicated the presence of Gal/Glc/GalNAc in a ratio of 2.8:1.0:0.9. Five permethylated alditol acetate peaks were characterized as 2,3,4,6‐OMe4Gal, 2,4,6‐OMe3Gal, 2,3,6‐OMe3Gal, 2,3,6‐OMe3Glc, and 4,6‐OMe2GalNAcMe by gas chromatography‐mass spectrometry. The anomeric carbohydrate sequence has been determined by specific exoglycosidase digestion. Six‐hundred megahertz 1H NMR spectroscopy of the oligosaccharide released by ceramide glycanase hydrolysis confirmed the structure of the Ngsl as Galβ1‐3GalNAcβ1‐3Galα1‐4Galβ1‐4Glc\1‐1Cer or IV3GalGbOse4Cer. Using the immunooverlay technique with anti‐stage‐specific embryonic antigen 3 antibody, it was found in bovine, rat, and normal adult human brain and bovine myelin, but not in human or rat myelin.

Chromatographic resolution and quantitative assay of CNS tissue sphingoids and sphingolipids

“…4 ) (27,33). In addition to the previously defined brain NGSL, a tetraglycosylceramide, GgOse 4 Cer (20,46); three pentaglycosylceramides, GbOse 5 Cer (47), Lewis X, and other long-chain NGSL of the Lewis X-series (27); and a novel GalNAc-GA1 ( 45) have been characterized. The CRAQ method will enable the characterization of several other, probably novel NGSL in CNS tissue that are revealed by DIG-IS.…”

Human Skin Cells That Express Stage-Specific Embryonic Antigen 3 Associate with Dermal Tissue Regeneration

“…8 In this initial report, Solter noted that SSEA3 was also expressed in erythrocytes (red blood cells), providing the first evidence that SSEA3 expression was not an exclusive cancer or stem/undifferentiated cell marker and may serve other functions in the body in a spatial, niche, and tissue-dependent manner. Reports of SSEA3-expressing cells within human tissues such as kidney, 18 brain 19 , and skin 10 have raised the possibility that the cell-surface expression of the SSEA3 epitope may facilitate the identification and isolation of ASC-like and/or undifferentiated cell subpopulations from adult human tissues. While SSEA3 has already been used to identify human skin-derived cells that are easier to reprogram to pluripotency 10,13 (suggesting a less differentiated state), associated with a multi-lineage differentiation potential, 11 (suggesting a multipotent or possibly pluripotent state) and correlated to the occlusion of human blood vessels 20 (suggesting a possible regeneration-associated state for blood vessels), SSEA3 has not been investigated as a cell-surface biomarker facilitating the identification and isolation of human skin cells directly associated with tissue regeneration after dermal injury.…”

Cell‐specific expression of neutral glycosphingolipids in vertebrate brain: Immunochemical localization of 3‐O‐acetyl‐sphingosine‐series glycolipid(s) in myelin and oligodendrocytes

“…Several NGSLs such as plasmalopsychosine, plasmalocerebrosides, lactosylceramide, globotriosylceramide, gangliotriosylceramide, globotetraosylceramide, lactotetraosylceramide, and fuconeolactotetraosylceramide or Lewis X (Le x ; Vanier et al, 1980; Ishikawa et al, 1987; Dasgupta et al, 1992; Nudelman et al, 1992; Levery et al, 1992) have been characterized in vertebrate brain. In recent years, additional NGSLs have been characterized (Dasgupta and Hogan, 1993; Dasgupta et al, 1995, 1996, 2000) as minor brain components and some of them have novel structures. These include gangliotetraosylceramide (GgOse 4 Cer, 2.5 mg/100 g acetone‐dried brain) or GA1 (Dasgupta et al, 1992; Dasgupta and Hogan, 1993), globopentaosylceramide (Galβ‐GbOse 4 Cer; Dasgupta et al, 1995), the Le x and its series (Dasgupta et al, 1996), GalNAc‐GA1 (1.25 mg/100 g acetone‐dried brain; Dasgupta et al, 2000), and the newly identified 3‐O‐acetylsphingosine series NGSLs (Dasgupta et al, 2002), which amount to approximately 20% (in total) of GalCer concentration in CNS and varies from species to species.…”

“…In recent years, additional NGSLs have been characterized (Dasgupta and Hogan, 1993; Dasgupta et al, 1995, 1996, 2000) as minor brain components and some of them have novel structures. These include gangliotetraosylceramide (GgOse 4 Cer, 2.5 mg/100 g acetone‐dried brain) or GA1 (Dasgupta et al, 1992; Dasgupta and Hogan, 1993), globopentaosylceramide (Galβ‐GbOse 4 Cer; Dasgupta et al, 1995), the Le x and its series (Dasgupta et al, 1996), GalNAc‐GA1 (1.25 mg/100 g acetone‐dried brain; Dasgupta et al, 2000), and the newly identified 3‐O‐acetylsphingosine series NGSLs (Dasgupta et al, 2002), which amount to approximately 20% (in total) of GalCer concentration in CNS and varies from species to species.…”

“…GA1, Le x , and GalNAc‐GA1 have been immunohistochemically localized in rodent brain, indicating that these NGSLs may have tissue and cell specificity in CNS, e.g., GA1 in myelinated fibers (Dasgupta and Hogan, 1993), Le x in the dendritic processes (Dasgupta et al, 1996), and GalNAc‐GA1 in neurons (Dasgupta et al, 2000). Gal‐GbOse 4 Cer has been purified and characterized in rat, bovine, and human brain (Dasgupta et al, 1995), but its localization in adult brain has not been determined, and this also applies to the 3‐O‐acetylsphingosine‐series NGSLs. Recent studies examining the expression of human and murine Gb 3 ‐ and Gb 4 ‐synthase genes (responsible for globo‐series GSL) indicate that β1,3GalNAcT (Gb 3 ‐synthase) is expressed weakly in murine brain but strongly in human brain (Okajima et al, 2000).…”