| Project/Area Number |
07807205
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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 |
Biological pharmacy
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| Research Institution | KYOTO SANGYO UNIVERSITY |
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Principal Investigator |
FUKUI Shigeyuki Dept.Biotechnology, Fac.Engineering, Kyoto Sangyo University, 工学部, 助教授 (30218888)
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| Co-Investigator(Kenkyū-buntansha) |
FUKUI Shigeyuki Dept.Biotechnology, Fac.Engineering, Kyoto Sangyo University (30218888)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1996: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1995: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | PC12 cells / neurite formation / poly-N-acetyllactosamine / nerve growth factor / 糖タンパク質 / 糖鎖 / ポリラグトサミン / 神経細胞 |
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| Research Abstract |
To explore the biological role of carbohydrate chains in relation to the process of nerve cell differentiation, I have carried out comparative study to characterize the changes of carbohydrate structure of cell surface glycoproteins by using conventional PC12 cells and its variant cells (PC12D). PC12D cells extend neurites faster than PC12 cells responding not only to nerve growth factor (NGF), but also to cyclic AMP.PC12D cells differ also morphologically from PC12 cells, being flat in shape and having expended short processes without any stimulation. To know whether the carbohydrate chains of cell surface glycoproteins of PC12 cells are alterd during neurite formation, and also differ from those of PC12D cells, I used the method of in vitro metabolic labeling of those cells by using either [^3H] glucosamine to label carbohydrate moieties, or [^3H] threonine to lable peptide portion of glycoproteins. In found that the size of carbohydrate chains obtained from PC12D cells was smaller in molecular weight in general than that of PC12 cells and the difference depended particularly on the length of poly-N-acetyllactosamine chain linked to a N-linked complex oligosaccharide core. Further, the NGF stimulation caused the decreased of poly-N-acetyllactosamine synthesis from PC12 cells while PC12D cells were insensitive in this regard.
From the results of the enzyme activity of glycosyltransferases which may participate in poly-N-acetyllactosamine synthesis, the decrease in the amount of the poly-N-acetyllactosamine chains in PC12D cells and NGF-primed PC12 cells was accounted for by the lowered activity of poly-N-acetyllactosamine extension enzyme (GnT-i), for the content of poly-N-acetyllactosamine chains in PC12 and PC12D cells correlated precisely with the changes in the GnT-i activity, whereas GlcNAc transferase V (GnT-V) and beta1-4 galactosyltransferase activities remained unchanged.
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