| Project/Area Number |
11680604
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
OKA Shogo Grad.Sch.Pharm.Sci., Kyoto U., Associate Professor, 薬学研究科, 助教授 (60233300)
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| Co-Investigator(Kenkyū-buntansha) |
KAWASAKI Toshisuke Grad.Sch.Pharm.Soci., Kyoto U., Professor, 薬学研究科, 教授 (50025706)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1999: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | HNK-1 epitope / glucuronyltransferase / cell adhesion molecule / gene-deficient mice / L1 / sulfotransferase / C6グリオーマ細胞 / 小脳 |
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| Research Abstract |
The HNK-1 carbohydrate epitope, which is recognized by monoclonal antibody HNK-1, is characteristically expressed on a series of cell adhesion molecules and also on some glycolipids in the nervous system over a wide range of species from insect to mammal. The HNK-1 epitope is involved in cell-cell and/or cell-substrate interaction and recognition during the development of the nervous system. The characteristic structural feature of this epitope is the sulfoglucuronyl residue, because the inner structure, Gal β1-4 GlcNAc, is found commonly in various glycoproteins and glycolipids, suggesting that glucuronyltransferase (s) and sulfotransferase (s) are key enzymes in the biosynthesis. We have recently cloned novel glucuronyltransferases (GlcAT-P and GlcAT-S) and a sulfotransferase cDNAs, which are key enzymes of the biosynthesis of the HNK-1 epitope. In the present study, we obtained following results using these cDNAs. 1) In situ hybridization analysis revealed that the different distributions of GlcAT-P and GlcAT-S were observed in each brain region in contrast to the ubiquitous expression of sulfotransferase. 2) Using the stable transformant of C6 glioma cells transfected with GlcAT-P cDNA, we demonstrated not only that the HNK-1 epitope was preferentially expressed on the NCAM and L1 molecules but also that the HNK-1 epitope expressed on L1 was mainly involved in the morphological changes of the cells. 3) To investigate the function of the HNK-1 epitope in vivo, we have generated the mice lacking GlcAT-P.We confirmed homologous recombination in GlcAT-P deficient mice by Southern blot analysis and absence of GlcAT-P expression by Northern blot analysis. Western blot analysis with HNK-1 antibody revealed that almost all of the HNK-1 carobhydrate epitope disappeared in the GlcAT-P deficient mice.
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