Carbohydrate antigen expression in murine embryonic stem cells and embryos. II. Sialylated antigens and glycolipid analysis.
A mouse embryonic stem (ES) cell line E14 and early mouse embryos were stained with a panel of 15 monoclonal antibodies recognizing sialylated or potentially sialylated carbohydrate determinants. Sialyl Le-x and sialyl Le-a were detected on the pre-implantation embryo from the 8-cell stage, and sialyl Le-a weakly on undifferentiated ES cells. Changes in cell surface carbohydrates occurred after induction of ES cell differentiation with retinoic acid (RA) and dibutyryl cAMP. Qualitative analysis of the neutral glycolipids of untreated and RA-treated ES cells using high-performance thin-layer chromatography (HPTLC) revealed few differences between the two types of culture. The major gangliosides in both cultures were indicative of an active 'a' ganglioside synthesis pathway. GD3, a precursor of the 'b' synthesis pathway, previously reported to be characteristic of embryonal carcinoma (EC) cells, was absent. RA-induced differentiation caused a shift in the spectrum to more complex gangliosides. Application of fast atom bombardment mass spectrometry (FAB-MS) to permethylated derivatives of individual bands permitted partial characterization of an unusual sialylated glycolipid and a rare ganglioside with the suggested structure of GalNAc-GD1a.
Kimber
S J
SJ
Department of Cell and Structural Biology, University of Manchester, UK.
Brown
D G
DG
Påhlsson
P
P
Nilsson
B
B
D000818:Animals; D001755:Blastocyst / Q000737:chemistry*; D018395:CA-19-9 Antigen; D002241:Carbohydrates / Q000032:analysis* / Q000276:immunology; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D005260:Female; D005732:Gangliosides / Q000032:analysis*; D006017:Glycolipids / Q000032:analysis*; D007150:Immunohistochemistry; D016256:Lewis X Antigen / Q000032:analysis; D051379:Mice; D008856:Microscopy, Fluorescence; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013194:Staining and Labeling; D013234:Stem Cells / Q000737:chemistry*
D018395:CA-19-9 Antigen; D002241:Carbohydrates; D005732:Gangliosides; D006017:Glycolipids; D016256:Lewis X Antigen; C064564:sialyl Le(a) ganglioside