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  1. 020 学位論文
  2. 生命科学研究科
  3. 20 生理科学専攻

Elucidation of mechanism that determines N-linked sugar chain pattern expressed in cells or tissues using cDNA macroarray and 2 dimensional HPLC analyses

https://ir.soken.ac.jp/records/1145
https://ir.soken.ac.jp/records/1145
ebb77ef1-be06-4073-bb4a-c1f0855df40b
名前 / ファイル ライセンス アクション
甲781_要旨.pdf 要旨・審査要旨 / Abstract, Screening Result (228.8 kB)
甲781_本文.pdf 本文 (4.6 MB)
Item type 学位論文 / Thesis or Dissertation(1)
公開日 2010-02-22
タイトル
タイトル Elucidation of mechanism that determines N-linked sugar chain pattern expressed in cells or tissues using cDNA macroarray and 2 dimensional HPLC analyses
タイトル
タイトル Elucidation of mechanism that determines N-linked sugar chain pattern expressed in cells or tissues using cDNA macroarray and 2 dimensional HPLC analyses
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 石井, 章寛

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石井, 章寛

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フリガナ イシイ, アキヒロ

× イシイ, アキヒロ

イシイ, アキヒロ

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著者 ISHI, Akihiro

× ISHI, Akihiro

en ISHI, Akihiro

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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781号
研究科
値 生命科学研究科
専攻
値 20 生理科学専攻
学位授与年月日
学位授与年月日 2004-03-24
学位授与年度
値 2003
要旨
内容記述タイプ Other
内容記述 N-linked sugar chains on glycoproteins are indispensable for the normal development. The expression of N-linked sugar chains is strictly regulated spatially and temporally, and tissue specific expression patterns of N-glycans are observed. Comparison of the mouse brain as well as the human brain revealed little variation among individuals of each species. However, molecular mechanisms that are responsible for the strict regulation of the N-linked sugar chains remain largely unknown. The expression of the N-glycans is based on the orchestrated action of many enzymes, glycosyltransferases and glycosidases, which catalyze biosynthesis and degradation of glycochains. The expression patterns of genes that encode these enzymes are, therefore, necessary to understand the regulatory mechanisms of N-glycan biosynthesis. In the present study, I developed a cDNA macroarray, with which expression of most of the glycosyltransferase and glycosidase genes can be analyzed at the same time. I chose this sensitive system because the expression levels of these genes were mostly too low to be analyzed by a microarray system. Using the cDNA macroarray system, I analyzed the gene expression patterns of more than 110 glycosyltransferases and glycosidases in the brain from 12-day mouse embryos, and in the brain, kidney, and liver from 12-week adult mice, and correlated them with the expression patterns of N-linked sugar chains in these tissues. The analyses revealed the tissue specific expression patterns of the glycosyltransferase and glycosidase genes as well as tissue specific N-glycan expression profiles. For example, whereas Golgi-mannosidase IB and polypeptide GalNAc Transferase I genes were equally expressed in these four tissues, α2, 8-sialyltransferase V and β1, 4-galactosyltransferase VI genes were highly expressed in the postnatal 12-week brain. mRNA amounts of some of the genes, which were differentially expressed in the four tissues were further verified by means of RT-PCR analyses. By comparing the gene expression and N-glycan expression profiles, I could obtain several new findings; bisecting structures were exclusively catalyzed by N-acetylglucosaminyltransferase III, and a core fucose is exclusively synthesized by fucosyltransferase VIII. In this study, I also compared the expression of the glyco-genes and N-linked sugar chains, and identified correlation between gene expression and N-linked sugar chains. For example, the gene expression of glucosidase I, glucosidase II and ER-mannosidase I was highly correlated Golgi-mannosidase IB with correlation coefficients (0.82, 0.74 and 0.80, respectively). Such coordinated expression could be necessary and important physiologically to biosynthesize the N-linked sugar chains smoothly and efficiently. cDNA macroarray system as well as high-throughput N-glycan analyzing system will provide significant information for the elucidation of the regulatory mechanisms and biological function of the N-linked sugar chains.
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