| Project/Area Number |
07680773
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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 |
Cell biology
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| Research Institution | KITASATO UNIVERSITY |
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Principal Investigator |
HOTTA Kyoko Kitasato Univ.School of Medicine, Dept.Biochem., Professor, 医学部, 教授 (10050402)
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| Co-Investigator(Kenkyū-buntansha) |
HORIUCHI Tadashi Daiichi Pharmaceutical Co., Basic Tech.Res.Lab., Head of Laboratory, 創薬基盤研究所, 研究所長
ICHIKAWA Takafumi Kitasato Univ.School of Medicine, Dept.Biochem., Research associate, 医学部, 助手 (30245378)
OHARA Susumu Kitasato Univ.School of Medicine, Dept.Biochem., Assistant Professor, 医学部, 講師 (90118795)
GOSO Yukinobu Kitasato Univ.School of Medicine, Dept.Biochem., Assistant Professor, 医学部, 講師 (20112659)
ISHIHARA Kazuhiko Kitasato Univ.School of Medicine, Dept.Chem., Professor, 医学部, 教授 (10104530)
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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 |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1996: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1995: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Hyaluronan / Gastric mucosa / Gastric epithelial cells / Transgenic mouse / SV40 / large T-antigen / GSM06 cells / ムチン / 粘液細胞 / 胃粘液 / モノクロナール抗体 / 胃表層粘液細胞 |
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| Research Abstract |
GSM06 is a cell line established from the stomach of transgenic mouse harboring a temperature-sensitive simian virus 40 (SV40) large T-antigen gene. ^3H-Labeled macromolecules produced by the cells incubated with [^3H] glucosamine were characterized to examine whether or not GSM06 cells synthesize mucin (mucus glycoprotein). The GSM06 cells grew until a confluent monolayr formed at 33ﾟC (the permissive temperature for SV40 large T-antigen expression), and the ^3H-labeled macromolecules appeared in both cell extract and medium during culture for at least one week. Unexpectedly, almost all ^3H-labeled macromolecules, which were excluded from a column of Sepharose CL-4B,were identified as hyaluronan by analyzes using Sepharose CL-2B chromatography, cesium trifluoroacetate equilibrium centrigugation, treatment with dithiothreitol, and trypsin and hyaluronidase digestion. At a nonpermissive temperature (39ﾟC), GSM06 cells grew only slightly, but produced much more hyaluronan than at 33ﾟC.The results indicate that GSM06 cells do not produce mucin, but hyaluronan, and that the expression of large T-antigen may influence hyaluronan synthesis in GSM06 cells.
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