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Carbohydrate Modification of 6B4 Proteoglycan, and Its Splicing Variant, Receptor-like Protein Tyrosine Phosphatase ζ in the developing chickbrain
https://ir.soken.ac.jp/records/1317
https://ir.soken.ac.jp/records/1317d96ba941-1b23-4591-b2c0-28574fd79534
名前 / ファイル | ライセンス | アクション |
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要旨・審査要旨 / Abstract, Screening Result (264.0 kB)
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本文 (7.0 MB)
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Item type | 学位論文 / Thesis or Dissertation(1) | |||||
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公開日 | 2010-02-22 | |||||
タイトル | ||||||
タイトル | Carbohydrate Modification of 6B4 Proteoglycan, and Its Splicing Variant, Receptor-like Protein Tyrosine Phosphatase ζ in the developing chickbrain | |||||
タイトル | ||||||
タイトル | Carbohydrate Modification of 6B4 Proteoglycan, and Its Splicing Variant, Receptor-like Protein Tyrosine Phosphatase ζ in the developing chickbrain | |||||
言語 | en | |||||
言語 | ||||||
言語 | eng | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_46ec | |||||
資源タイプ | thesis | |||||
著者名 |
濱中, 裕喜
× 濱中, 裕喜 |
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フリガナ |
ハマナカ, ヒロキ
× ハマナカ, ヒロキ |
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著者 |
HAMANAKA, Hiroki
× HAMANAKA, Hiroki |
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学位授与機関 | ||||||
学位授与機関名 | 総合研究大学院大学 | |||||
学位名 | ||||||
学位名 | 博士(理学) | |||||
学位記番号 | ||||||
内容記述タイプ | Other | |||||
内容記述 | 総研大甲第236号 | |||||
研究科 | ||||||
値 | 生命科学研究科 | |||||
専攻 | ||||||
値 | X2 分子生物機構論専攻 | |||||
学位授与年月日 | ||||||
学位授与年月日 | 1996-09-30 | |||||
学位授与年度 | ||||||
値 | 1996 | |||||
要旨 | ||||||
内容記述タイプ | Other | |||||
内容記述 | The brain contains a diverse set of proteoglycans as the major component of the cell-surface glycoconjugates and of the extracellular matrix (ECM). 6B4 proteoglycan (6B4 PG) is one of the major soluble chondroitin sulfate proteoglycans in the brain, and has a structure corresponding to the extracellular domain of a proteoglycan-type protein tyrosine phosphatase, PTPC, (RPTPβ). In this study, I first identified chick 6B4 PG by purifying it from embryonic chick brain and determining the N-terminal amino acid sequence. The obtained sequence corresponded well to that of rat 6B4 PG. From the biochemical properties of the purified protein and its immunoreactivity with MAb 6B4, I concluded that MAb 6B4 (originally raised against rat 6B4 PG)-reactive proteoglycan in chick tissue is chick 6B4 PG.<br /> Next, I prepared polyclonal antibodies which specifically recognize chick 6B4 PG and PTPζ, and analyzed the carbohydrate structures on the two molecules in the developing chick brain. Immunoprecipitation experiments using these antibodies revealed that almost all of the keratan sulfate and protein-bound Lex carbohydrate structure in the brain were exclusively bound to 6B4 PG and PTPζ. The modifications with keratan sulfate and Lex carbohydrate occurred on both of 6B4 PG and PTPζ. Furthermore, I found that the modifications of 6B4 PG and PTPζ with these carbohydrates are spatiotemporally regulated during development.<br /> Addition of keratan sulfate to these proteoglycans markedly increased from embryonic day 11 (E11), in contrast to that of Lex and HNK-1 carbohydrates which increased gradually during development in accordance with expression of the core proteins. This suggests that keratan sulfate plays some specific roles in development. In the developing tectum, in contrast to the uniform distribution of the core proteins during development, keratan sulfate was expressed only in the lateral region of the tectum at E10, and the expression was extended toward the dorsal area until E13. Moreover, keratan sulfate modification occurs in several restricted regions of the early embryonic chick brain, especially at morphogenetic boundary regions such as the roof plate of the tectum, the zona limitans intrathalammca in the diencephalon, and the mes-metencephalic boundary.<br /> At the mes-metencephalic boundary, keratan sulfate modification of these proteoglycans was specifically observed from E3 to E6 on a ring of cells encircling the neural tube. In this region, keratan sulfate staining was observed on the cell bodies and the radially oriented processes. These processes seem to be stained with anti-vimentin antibody but not with anti-neurofilament antibody. Therefore, I tentatively defined these processes as radial glial fibers that are considered to be the scaffold for migrating neurons. I also found that the neurofilament- and Ng-CAM-positive growing axons passed through the keratan sulfate-rich mes-metencephalic boundary region. The expression pattern of keratan sulfate in this region spatiotemporally corresponded well to the Wnt-1 and Fgf-8 expression and to the fovea isthmi formation, a groove separating the mes-metencephalon along the neural tube. I found that these locally expressed keratan sulfate is also bound to 6B4 PG and PTP9. These results suggest that keratan sulfate on 6B4 PG and PTPζ plays important roles in brain morphogenesis by modulating the cell-cell and/or cell-substratum interactions mediated by these molecules. | |||||
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値 | 有 | |||||
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内容記述タイプ | Other | |||||
内容記述 | application/pdf |