Molecular Biology of Differentiation of Tooth Forming Cells

牙齿形成细胞分化的分子生物学

• 批准号: 63480410
• 负责人: SHIMOKAWA Hitoyata
• 金额: $ 3.71万
• 依托单位: TOKYO MEDICAL AND DENTAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63480410/
• 关键词: Enamel; Dentin; Mineralization; Amelogenin; Osteonectin; Differentiation; リンタンパク質

1. Immunochemical and immunohistochemical localization of osteonectin.Various tissues were investigated for the presence of osteonectin by means of immunochemical and immunohistochemical methods. Osteonectin could be demonstrated in active osteoblasts and osteoprogenitor cells as well as in young osteocytes. Osteonectin expression, however, was not confined to mineralizing tissue, but was distributed in many soft tissues.2. cDNA cloning of osteonectin from expression libraries of bovine odontoblast.To examine the identity of osteonectin with SPARC, cloning of osteonectin cDNA from lambda gtll expression library of bovine odontoblasts (non-cultured cells) was carried out. A 2.lkbp cDNA coding for osteonectin, isolated from the librafy, was sequenced. The nucleotide sequence predicted that osteonectin contained 304 amino acids including 17-residue signal peptide. The osteonectin sequence showed near identity (>90%) with another protein, SPARC.3. Expression of SPARC/osteonectin transcript.Osteonectin cDNA was used to analyze the expression of the corresponding transcript in various bovine fetal tissues. Several non-botie tissues, including skin, muscle, tendon, expressed the transcript in lesser extent than bone tissue.4. Factors effect on the expression of osteonectin.It was found that mRNA for alpha2(1) collagen and osteonectin were markedly increased in TGF-,beta -treated MG-63 and HOS human osteosarcoma cells. The messages of these proteins also increased in 1,25(OH)_2D_3 treated cells.5. Human amelogenin gene.Bovine amelogenin cDNA was used to isolate the amelogenin gene from a library of human genomic DNA fragments which contained a part of amelogenin coding region spanning approximately 13 kilobases. Analysis of this gene indicated the presence of four exons at least. It was revealed that the human amelogenin had a high homology with bovine, porcine, and murine amelogenins and had also hydrophilic carboxyterminus telopeptide.

1.骨连接蛋白的免疫化学和免疫组织化学定位。通过免疫化学和免疫组织化学方法研究各种组织中骨连接蛋白的存在。骨连接素可以在活跃的成骨细胞和骨祖细胞以及年轻骨细胞中得到证实。但骨连接素的表达并不局限于矿化组织，而是分布在多种软组织中。2．从牛成牙本质细胞表达文库克隆骨连接蛋白的cDNA。为了用SPARC检查骨连接蛋白的同一性，从牛成牙本质细胞(非培养细胞)的lambda gtll表达文库克隆骨连接蛋白cDNA。对从文库中分离出的编码骨连接蛋白的 2.lkbp cDNA 进行了测序。核苷酸序列预测骨连接蛋白含有304个氨基酸，其中包括17个残基的信号肽。骨连接蛋白序列显示与另一种蛋白质 SPARC.3 接近同一性 (>90%)。 SPARC/骨连接素转录本的表达。利用骨连接素cDNA分析相应转录本在各种牛胎儿组织中的表达。一些非机器人组织，包括皮肤、肌肉、肌腱，其转录物的表达程度低于骨组织。4．影响骨连接蛋白表达的因素。发现TGF-β处理的MG-63和HOS人骨肉瘤细胞中α2(1)胶原和骨连接蛋白的mRNA显着增加。这些蛋白质的信息在 1,25(OH)_2D_3 处理的细胞中也有所增加。5。人牙釉蛋白基因。牛牙釉蛋白cDNA用于从人类基因组DNA片段文库中分离牙釉蛋白基因，该文库含有跨越约13kbp的牙釉蛋白编码区的一部分。对该基因的分析表明至少存在四个外显子。结果表明，人牙釉蛋白与牛、猪和鼠的牙釉蛋白具有高度同源性，并且还具有亲水性羧基末端端肽。

项目成果

Y.Ogata: "Calcium Binding Activity of Bovine Enamelin." Dentistry in Japan. 26. 51-55 (1989)

Y.Ogata：“牛牙釉质的钙结合活性。”

H. Shimokawa: Tooth Enamel V. Florence Publishers, 545 (1989)

H. Shimokawa：牙釉质 V. 佛罗伦萨出版社，545 (1989)

下川仁弥太,荊木京未,鈴木ミチ子,田村正人,佐々木哲,森山啓司,下川伶子,土井豊: 日本骨代謝学会雑誌. 6(1). 34-40 (1988)

Jinya Shimokawa、Kyomi Aki、Michiko Suzuki、Masato Tamura、Tetsu Sasaki、Keiji Moriyama、Reiko Shimokawa、Yutaka Doi：日本骨代谢学会杂志 6(1) (1988)。

H. Shimokawa: "Studies on Expression of Osteonectin" Jap. J. Bone Metab. 7: (1) 21-25, 1989.

H. Shimokawa：“骨连接素表达的研究”Jap。

Y.Doi: "Osteonectin Inbiting De Novo Formation of Apatite in the Presence of Collagen." Calcif.Tissue Int.44. 200-208 (1989)

Y.Doi：“骨连接素在胶原蛋白存在下抑制磷灰石的重新形成。”