Ganglioside Composition of Normal and Mutant Mouse Embryos

Abstract

The enrichment of gangliosides in neuronal membranes suggests that they play an important role in CNS development. We recently found a marked tetrasialoganglioside deficiency in twl/twl mutant mouse embryos at embryonic day (E)-11. The recessive twl/twl mutants die at embryonic ages E-9 to E-18 from failed neural differentiation in the ventral portion of the neural tube. In the present study, we examined the composition and distribution of gangliosides in twl/twl mutant mouse embryos at E-12. The total ganglioside sialic acid concentration was significantly lower in the mutants than in normal (+/-) embryos. The mutants also expressed significant deficiencies of gangliosides in the "b" metabolic pathway (GD3, GD1b, GT1b, and GQ1b) and elevations in levels of gangliosides in the "a" metabolic pathway (GM3, GM2, GM1, and GD1a). These findings suggest that the mutants have a partial deficiency in the activity of a specific sialyltransferase in the b pathway. Regional ganglioside distribution was also studied in E-12 normal mouse embryos. The ganglioside composition in heads and bodies was similar to each other and to whole embryos. Total ganglioside concentration and the distribution of b pathway gangliosides were significantly higher in neural tube regions than in nonneural tube regions. These findings suggest that b pathway gangliosides accumulate in differentiating neural cells and that the deficiency of these gangliosides in the twl/twl mutants is closely associated with failed neural differentiation.

Roles of gangliosides in mouse embryogenesis and embryonic stem cell differentiation

“…Bouvier and Seyfried (1989) observed that the predominant gangliosides in E12 mouse embryos were GD3 (51% of total sialic distribution), GM3 (19%), and GT1b (9.6%); other gangliosides occurred in much lower amounts (GM2 (2.6%), GM1 (1.6%), GD1a (3.7%), GD1b (6.3%), and GQ1b (4.5%)). Similar distributions were observed in both neural and non-neural embryonic structures, suggesting that undifferentiated embryonic cells in mice express GM3 and GD3 as the major ganglioside species (Bouvier and Seyfried, 1989). However, little is known about the expression of GT1b in preimplantation embryos.…”

“…On E13, mST3GalV mRNA was expressed in various neural and non-neural tissues and in the telencephalon, while on E15, strong expression of mST3Gal V was observed in the liver (Ji et al, 2000). Bouvier and Seyfried (1989) observed that the predominant gangliosides in E12 mouse embryos were GD3 (51% of total sialic distribution), GM3 (19%), and GT1b (9.6%); other gangliosides occurred in much lower amounts (GM2 (2.6%), GM1 (1.6%), GD1a (3.7%), GD1b (6.3%), and GQ1b (4.5%)). Similar distributions were observed in both neural and non-neural embryonic structures, suggesting that undifferentiated embryonic cells in mice express GM3 and GD3 as the major ganglioside species (Bouvier and Seyfried, 1989).…”

“…Studies have reported that there are drastic changes in the expression patterns and levels of gangliosides during embryonic development. The changes occur in the gangliosides themselves (Yu et al, 1988;Bouvier and Seyfried, 1989), as well as in the glycosyltransferases and glycosidases (Ishii et al, 2007) that regulate ganglioside synthesis. Other recent studies based on analyses in genetically engineered animals, have demonstrated that gangliosides mainly play roles in the maintenance and repair of nervous tissues (Furukawa et al, 2007;Kittaka et al, 2008).…”

“…Cell surface molecules that can be used as markers for the identification and isolation of stem cells are essential for basic biological study and clinical use of ES cells. Glycolipids on the cell surface can serve as marker molecules GM3, GM1, GD1a, GM2, GT1b, GD3 Ji et al, 2000Yu et al, 1998E12 GD3, GM3, GT1b, GM2, GM1, GD1a, GD1b, GQ1b Ji et al, 2000Yu et al, 1998Bouvier& Seyfried, 1989E13 GD3, GM3, GT1b, GM2, GM1, GD1a, GD1b, GQ1b Ji et al, 2000Yu et al, 1988E14 GD3, GM3, GM2, GM1, GD1a, GT1b Nakamukote et al, 2007Bouvier & Seyfried, 1989 GM3,GM1,GD1a,GD3 In this review, we will describe the gangliosides expressed during mouse ovulation, spermatogenesis, and embryogenesis, as well as in stem cells, and discuss their availability as biomarkers for the identification of mES cells and their differentiation. Figure 1 shows the metabolic pathways for ganglioside production in the mouse.…”

Expression cloning of a CMP-NeuAc:NeuAc alpha 2-3Gal beta 1-4Glc beta 1-1'Cer alpha 2,8-sialyltransferase (GD3 synthase) from human melanoma cells.

“…Flow cytometric analysis, enzyme assay, and product characterization showed that the enzyme catalyzes the formation of GD3 by the transfer of a sialic acid molecule from CMP-NeuAc to the terminal sialic acid of GM3 via an a2-8 linkage. A GD3 synthase is a key enzyme in the "b" metabolic pathway of gangliosides (GD3, GD1b, GT1b, and GQ1b) which are associated with neural cell differentiation (44)(45)(46)(47).…”