The polymorphic carbohydrate structures of gangliosides play regulatory roles. In particular, b-series gangliosides, all of which contain ␣-2,8 sialic acids, have been considered to be critical in various biological events such as adhesion, toxin binding, neurite extension, cell growth, and apoptosis. To clarify the physiological functions of b-series gangliosides in vivo, we have established a gene knockout mouse of GD3 synthase. Although all b-series structures were deleted in the mutant mice, they showed an almost complete nervous tissue morphology with no apparent abnormal behavior. Moreover, no differences in Fas-mediated apoptotic reaction of lymphocytes between wild type and the mutant mice were detected. However, the mutant mice exhibited clearly reduced regeneration of axotomized hypoglossal nerves compared with the wild type, suggesting that b-series gangliosides are more important in the repair rather than in the differentiation of the nervous system and apoptotic process induced via Fas.Gangliosides, sialic acid-containing glycosphingolipids, are enriched in the nervous tissues of mammals and birds, and also distributed in various tissues and cells (1), playing regulatory roles with their polymorphic carbohydrate structures. In particular, b-series gangliosides including GD3, 1 GD2, GD1b, GT1b, and GQ1b have been considered to be critical as receptors for bacteria toxins (2) and are adhesion-associated (3, 4), neurite-inducing (5), cell growth-promoting (6, 7) and apoptosis-mediating molecules (8).To clarify the physiological functions of these b-series gangliosides in vivo, we have established a gene knockout mice line for ␣-2,8-sialyltransferase (9), which is responsible for the generation of all b-series gangliosides as well as c-series gangliosides. As expected, all b-series structures were deleted in the mutant mice, and a-series species were slightly increased instead. However, these b-series ganglioside-lacking mice showed an almost intact morphology of the brain and other nervous tissues, and no clear abnormal behaviors were detectable during the early period of life. Despite that GD3 has been considered to mediate Fas-induced apoptosis in lymphoid cells (8,10), no differences in the apoptotic reaction of lymphocytes between wild-type and mutant mice were detected. However, the mutant mice exhibited clearly reduced regeneration of axotomized hypoglossal nerves compared with the wild type, suggesting that b-series gangliosides are important in the repair of damaged nerves rather than in the differentiation of the nervous system.
EXPERIMENTAL PROCEDURESGeneration of GD3 Synthase Gene Knockout Mice-The chromosomal GD3 synthase gene was isolated from the gt11 phage library using GD3 synthase cDNA (pD3T-31) and mapped as described previously (11). To distinguish the true GD3 synthase gene from pseudo-genes, in situ hybridization was performed, and the identity was confirmed based on the correspondence of the gene assignment between humans and mice. The neo r gene was inserted between the BalI and ...