歯胚発育障害における基底膜型ヘパラン硫酸プロテオグリカンの役割

基底膜型硫酸乙酰肝素蛋白多糖在牙胚发育障碍中的作用

• 批准号: 13771071
• 负责人: 依田 浩子
• 金额: $ 1.28万
• 依托单位: Niigata University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13771071/
• 关键词: 基底膜型ヘパラン硫酸プロテオグリカン; パールカン; 歯胚; エナメル器; 物質輸送; 上皮内基質; エナメル上皮腫; 細胞増殖; ヘパラン硫酸プロテオグリカン; 歯胚発育障害; アンチセンス; 器官培養

今年度は前年に引き続き、正常歯胚組織の発育過程におけるin vivoでの基底膜型ヘパラン硫酸プロテオグリカン・パールカン(HSPG)の局在を明らかにする目的で,各歯胚発育段階のマウス下顎第一臼歯歯胚(胎生11,13,15,17日齢,生後1,6,10,12,28日齢)について,パラフィン連続切片をもちいて,HSPGコア蛋白質の局在およびHSPG遺伝子発現を免疫組織化学的およびin-situハイブリダイゼーション法にて検索した。その結果,HSPGコア蛋白質は,エナメル器および歯乳頭組織に局在し,とくに,エナメル髄星状網細胞の細胞間隙にきわめて豊富であった。鐘状期歯胚では,エナメル基質形成開始期に限局して,内エナメル芽細胞の細胞基底側および側面の細胞間隙にHSPG免疫陽性の小胞状拡張が出現し,周囲の内エナメル髄細胞にHSPG遺伝子発現がみられた。同時に,象牙芽細胞にもHSPG遺伝子発現が強く認められた。したがって,HSPGがそれらの細胞分化あるいはエナメル髄内の拡散による物質輸送の担体としてエナメル質・象牙質基質合成に関与している可能性が示唆された(裏面研究発表2-4)。また,歯胚組織由来とされるエナメル上皮腫においても,エナメル髄様の腫瘍胞巣内にHSPGが特異的高濃度に局在するとともに,腫瘍胞巣浸潤先端部でとくに遺伝子発現が強調されたので,上皮性腫瘍細胞の増殖や浸潤を制御している可能性が示唆された(研究発表1)。ついで,歯胚の各構成細胞でのHSPGコア蛋白質の生合成を確認する目的で,歯小嚢・歯乳頭・エナメル髄細胞の初代培養システムを確立し,蛍光抗体法および免疫沈降法,さらにRT-PCR法によって検討したところ,すべての細胞種においてHSPGの蛋白質およびmRNAレベルでの発現が確認された(研究発表4)。最後にHSPGの関与する歯胚発育障害機構を解明する目的で,アンチセンスHSPGオリゴヌクレオチド添加・非添加下にて歯胚器官培養をおこなった。アンチセンス添加群の培養歯胚紐織では,エナメル器の発育および象牙芽細胞分化が不良であることが確認されており,HSPGがエナメル芽細胞および象牙芽細胞分化に重要な役割をはたしていることがin vitroでも明らかになった。

This year, the year before last, the normal embryo tissue was introduced to the process of embryo rearing, and the normal embryo tissue was incubated. The basement membrane type of in vivo was used in the process of maturation. This year, the normal embryo tissue and the normal embryo tissue were introduced in the year before last, the normal embryo tissue and the normal embryo tissue in the year before last, the normal embryo tissue and the normal embryo tissue in the year before last, the normal embryo tissue and the normal embryo tissue in the year before last, the normal embryo tissue, normal embryo tissue, normal embryo The immuno-histochemical staining of HSPG protein was detected in the HSPG gene. The immunocytochemical method was used to detect the immunocytochemistry of the protein. The results showed that the HSPG protein was detected, and the milk tissue was detected in the breast tissue. The stellate cell gap was rich. In the early stage of embryonic development, there is a limited cycle in the initial stage, the basal cell surface of the basal cell, the cell gap of HSPG, the immune cell of the small cell, and the HSPG cell of the cell in the periphery. At the same time, the ivory buds show strong HSPG activity. In the case of HSPG, the possibility of biosynthesis and biosynthesis of ivory is indicated by the possibility of cell differentiation (Table 2-4). The origin of the embryo tissue is different from that of the epithelium. in this way, the high degree of HSPG in the cells is very high, and the cells are impregnated at the end of the cell to show the strength of the cells, and the possibility of impregnation of the epithelial cells indicates the possibility of cell infiltration. (table 1). The cells and embryos are formed into cells, HSPG, protein, protein and protein. The HSPG protein was tested and the mRNA was confirmed (Table 4 of the study). In the end, the HSPG machine and the embryo incubation mechanism were used to understand the purpose of the disease, so that the HSPG organ culture system was added. Add a group of germ cells to the tissue culture, which is used to breed the ivory germ cells to differentiate into poor cells, to confirm that the ivory buds differentiate badly, to HSPG the ivory buds to differentiate into cells, to make sure that the ivory buds are differentiated, and to cut the ivory buds for important purposes. To cut the ivory buds, to make sure that the ivory buds are differentiated, and to make sure that the ivory buds are not differentiated properly.

项目成果

Hiroko Ida-Yonemochi, et al.: "The basement membrane-type heparan sulfate proteoglycan (perlecan) in ameloblastomas : its intercellular localization in stellate reticulum-like foci and biosynthesis by tumor cells in culture"Virchows Archiv. 441. 165-173 (

Hiroko Ida-Yonemochi 等人：“成釉细胞瘤中的基底膜型硫酸乙酰肝素蛋白聚糖（基底膜聚糖）：其在星状网状病灶中的细胞间定位以及培养物中肿瘤细胞的生物合成”Virchows Archiv。

Hiroko Ida-Yonemochi, et al.: "Disturbed tooth eruption in osteopetrotic (op/op) mice : histopathogenesis of tooth malformation and odontomas"Journal of Oral Pathology & Medicine. 31. 361-373 (2002)

Hiroko Ida-Yonemochi 等人：“骨石症 (op/op) 小鼠牙齿萌出紊乱：牙齿畸形和牙瘤的组织病理学”口腔病理学杂志

Hiroko Ida-Yonemochi, et al.: "No developmental failure of cultured tooth germs from osteopetrotic (op/op) mice"Journal of Oral Pathology & Medicine. 31. 374-378 (2002)

Hiroko Ida-Yonemochi 等人：“骨石症 (op/op) 小鼠培养的牙胚未出现发育障碍”口腔病理学杂志

依田 浩子: "マウス歯胚発育過程における基底膜型ヘパラン硫酸プロテオグリカンの局在"歯科基礎医学会雑誌. 44・5. 104 (2002)

与田裕子：“小鼠牙胚发育过程中基底膜型硫酸乙酰肝素蛋白多糖的定位”日本基础牙科医学会杂志44・5.104（2002年）。

Hiroko-Ida-Yonemochi: "The basement membrane-type heparan sulfate proteoglycan (perlecan) in ameloblastomas : its intercellular localization in stellate reticulum-like foci and biosynthesis by tumor cells in culture"Virchows Archiv. 438. s00428-001-s00428

Hiroko-Ida-Yonemochi：“成釉细胞瘤中的基底膜型硫酸乙酰肝素蛋白聚糖（基底膜聚糖）：其在星状网状病灶中的细胞间定位以及培养物中肿瘤细胞的生物合成”Virchows Archiv。