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標的遺伝子組換えによるガラクトシルトランスフェラーゼ遺伝子の発生過程での機能解析

通过靶向基因重组对发育过程中的半乳糖基转移酶基因进行功能分析

基本信息

批准号：
07780649
负责人：
浅野雅秀
金额：
$ 0.64万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Encouragement of Young Scientists (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至无数据
项目状态：
已结题

项目摘要

Asnに結合する複合型糖鎖のガラクトース(Gal)の転移を担い、それ自身も細胞接着分子として機能することが示唆されているβ1, 4-ガラクトース転移酵素(β1, 4-GalT)遺伝子を欠損したマウスを作製して、生体内での糖鎖の機能を解析した。昨年度までの研究により、β1, 4-GalT遺伝子に変異を導入したES細胞クローンを9個単離していたので、本年度は3クローンについて集合キメラ法によりキメラマウスの作製を行ったところ、2クローンが生殖系列に伝達され、目的のβ1, 4-GalT遺伝子欠損マウスの作製に成功した。ホモ欠損マウスではβ1, 4-GalT活性がほとんど検出されず、レクチンブロット解析からもほとんどの糖タンパク質の糖鎖はGalが結合しておらず、GlcNAcで糖鎖の転移が止まっていることが確かめられた。ホモ欠損マウスは正常に生まれてくるが、生後数日から著しい成長遅延と激しい皮膚の角質化が認めれ、離乳前に約半分のホモ欠損マウスが死亡し、4カ月齢では80%のマウスが死亡した。各臓器の組織切片を調べたところ、皮膚の基底細胞層が肥厚し、小腸のクリプトが巨大化しており、これらの領域に存在する上皮幹細胞の増殖が亢進していることが示された。実際、小腸のクリプトでの細胞増殖速度が正常マウスに比べて約4倍に亢進しており、β1, 4-GalTにより合成される糖鎖は上皮細胞の増殖を負に制御していることが明らかとなった。また二糖類分解酵素の発現を指標に離乳前の小腸上皮細胞の分化状態を調べたところ、ホモ欠損マウスではラクターゼの発現が低く、逆に離乳後に発現してくるマルターゼやイソマルターゼが既に発現していることがわかった。表皮上皮細胞の増殖の増大は角化亢進の原因であり、小腸上皮の分化異常、特にラクターゼの欠損は離乳前の成長遅延と死亡の原因と考えられる。以上のようにβ1, 4-GalTの合成する糖鎖は上皮細胞の増殖と分化を制御しているという全く新しい知見が得られた。今後はこのマウスを用いることにより、初期胚の発生や細胞の癌化における糖鎖の役割についても新しい知見が得られることが期待できる。なお以上の結果は現在論文を投稿中である。

Asn was combined with recombinant glycosyltransferase (Gal) gene transfer assay, and autosomal cell culture followed by molecular assay was used to demonstrate the expression of β 1, 4-glucosyltransferase (β 1, 4-GalT) gene. In vivo glucose assay was performed. In last year's study, the children of, β 1, and 4-GalT were involved in the study of 9 families in the ES cell. This year, the collection of drugs was used in the study, and the reproductive series was completed in the second year. The goal was β 1, and the success of the 4-GalT program was not successful. In the absence of 4-GalT activity, there is no significant difference between the two groups. In this case, we need to know how to make sure that you are not aware of the situation. You need to know that you have a problem. You need to make sure that you have a problem with the combination of sugar, Gal, and GlcNAc, so that you can make sure that you are in trouble. In the first few days of his life, he was in the process of growing up, the skin of the baby, the half of the baby, the death of the baby, and the death of the baby. The tissue sections of each organ show signs of hypertrophy of the basal cells of the skin, hypertrophy of the basal cells of the skin, hypertrophy of the epithelial cells, hypertrophy of the epithelial cells, hypertrophy of the basal cells of the skin, hypertrophy of the basal cells of the skin. The rate of cell proliferation is about 4 times higher than that of normal cells. β 1, 4-GalT is used to synthesize sugar, epithelial cells, cells, and control cells. Disaccharide proteolytic enzymes indicate that the differentiation of small epithelial cells in front of the milk is in a state of differentiation, that is, the differentiation of the small epithelial cells in front of the milk, the differentiation of small epithelial cells in the milk, the differentiation of the small epithelial cells before the milk, the differentiation of the small epithelial cells in front of the milk, the differentiation of the small epithelial cells in front of the milk, the differentiation of the epithelial cells, the differentiation of the small epithelial cells, the differentiation of the small epithelial cells before the milk. The causes of hyperkeratosis of epidermis, differentiation of epithelium, growth and death of epidermis have been studied. The synthesis of sugar, the differentiation of epithelial cells, the differentiation of cells, the synthesis of β 1, the synthesis of β 1, the synthesis of glucose, the differentiation of epithelial cells, the differentiation of cells, the synthesis of β 1, the synthesis of sugar, the synthesis of sugar, the differentiation of epithelial cells, the differentiation of colonies, the synthesis of glucose, the synthesis of glucose, the differentiation of epithelial cells, the differentiation of colonies, the control of differentiation, the synthesis of glucose, the synthesis of glucose, the differentiation of epithelial cells, the differentiation of colonies, the control of differentiation, the synthesis of glucose, the synthesis of glucose, the differentiation of epithelial cells, the differentiation of colonies, the formation of new information. In the future, we will tell you that in the early stage of the embryo, you will get a lot of cancer, and the sugar will be cut in order to get new information. The results of the above results are now in the submission of the article.