FUNCTIONAL ANALYSIS OF TOOTH GERM DEVELOPMENT-RELATED GENES

牙胚发育相关基因的功能分析

• 批准号: 13470384
• 负责人: SAKAI Hidetaka
• 金额: $ 9.15万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13470384/
• 关键词: tooth germ; generation and regeneration; cDNA subtraction; in situ hybridization; antisense S-ODN; organ culture; purification of stem cell; yeast two hybrid; 幹細胞純化; 発生と再生; 幹細胞純化法

We previously examined the development of the mouse mandible, and demonstrated that odontogenesis occurs between embryonic day 10.5 (E 10.5) and E12. Based on the histological findings, we performed cDNA subtraction between the E10.5 and E12 mandibles to detect any differentially expressed genes which might be involved in the initiation of odontogenesis. We found Pgk-1,Ccte,Hsp86,Nucleolin,Hsc73,Frg1,N-ras,Set alpha and Hsj2 from the E10.5 mandible, and E25,ATPase6,Mum2, Thymosin beta4 and L21 from the E12 mandible to be differentially expressed genes. In situ hybridization analyses showed the expression of Set alpha, heat shock proteins (HSPs:Hsc73,Hsj2 and Hsp86), Pgk-1,Nucleolin, ATPase6, Thymosin beta4 to be detected in the developing tooth germ, thus indicating a close relationship of this gene to odontogenesis.Runx2/Cbfal knockout mice showed both a complete lack of ossification and the developmental arrest of tooth germ. We examined Runx2/Cbfa1 isoform-type specific functional r … More oles in the development of tooth germ by the administration of antisense phosphorothioate oligodioxynucleotides (S-ODNs) into cultured mouse mandibles. The administration of type II/III Runx2/Cbfa1 antisense S-ODNs into the culture media resulted in an arrest of tooth germ growth at the bud-like stage in cultured mandible taken from the 11-day-old embryos, while also causing the inhibition of the differentiation of odontogenic cells into ameloblast and odontoblast in cultured tooth germs taken from the 15-day-old embryos. The expression of dentin matrix protein 1, dentin sialophosphoprotein, amelogenin and ameloblastin was shown to be markedly suppressed in cultured tooth germ by the semi-quantitative RT PCR method. Meanwhile, no developmental arrest of tooth germ, no inhibition of gene expression, or differentiation of odontogenic cells was observed in samples treated with the type I Runx2/Cbfal antisense S-ODNs. The same findings were also observed in either the control or the sense and random sequence S-ODNs treated samples. These data indicate that the type II/III Runx2/Cbfa1 isoform is closely related to the development and differentiation of tooth germ. Less

我们之前研究了小鼠下颌骨的发育，并证明牙齿发育发生在胚胎第10.5天(E10.5)到E12天之间。根据组织学结果，我们对E10.5和E12下颌骨进行了cDNA消减，以检测可能与牙体发育启动有关的差异表达基因。在E10.5下颌骨中发现了PGK-1、Ccte、Hsp86、核仁素、Hsc73、Frg1、N-ras、SETα和Hsj2等基因，在E12下颌骨中发现了差异表达基因E25、ATPase6、Mum2、Thymosin beta4和L21。原位杂交分析表明，SETα、热休克蛋白(Hsps：Hsc73、Hsj2和Hsp86)、PGK-1、核仁素、ATPase6、胸腺素β4在发育中的牙胚中均有表达，表明该基因与牙胚的发生密切相关。我们检测了Runx2/Cbfa1异构型特异性功能r…通过向培养的小鼠下颌骨中注入反义硫代寡核苷酸(S-ODN)来促进牙胚的发育。在培养的11d胚胎的下颌骨中加入II/III型Runx2/Cbfa1反义S寡核苷酸后，牙胚的生长受阻于芽状期，同时也抑制了15d胚胎培养的牙胚向成釉细胞和成牙本质细胞的分化。半定量逆转录聚合酶链式反应显示牙本质基质蛋白1、牙本质涎磷蛋白、成釉蛋白和成釉蛋白在体外培养的牙胚中的表达受到明显抑制。同时，在I型Runx2/Cbfal反义S-ODN处理的样本中，没有观察到牙胚发育停滞、基因表达抑制或牙源性细胞分化。在对照组和正义和随机序列S-ODN处理的样本中也观察到了同样的发现。这些数据表明，II/III型Runx2/Cbfa1亚型与牙胚的发育分化密切相关。较少

项目成果

Haruyoshi Yamaza, et al.: "Detection of differentially expressed genes in the early developmental stage of the mouse mandible"International Journal of Developmental Biology. 45・4. 675-680 (2001)

Haruyoshi Yamaza等：“小鼠下颌骨早期发育阶段差异表达基因的检测”国际发育生物学杂志45・4（2001）。

An Li element disrupts human bone sialoprotein promoter : lack of tissue-specific regulation by distalless5 (Dlx5) and runt homeodomain protein2 (Runx2)/core binding factor al (Cbfal) elements

Li元件破坏人骨唾液酸蛋白启动子：缺乏远端5（Dlx5）和矮同源域蛋白2（Runx2）/核心结合因子al（Cbfal）元件的组织特异性调节

• 发表时间: 2002
• 期刊: Gene. 299
• 作者: Tamotsu Kiyoshima;et al.
• 通讯作者: et al.

Detection of differentially expressed genes in the early developmental stage of the mouse mandible.

小鼠下颌骨发育早期差异表达基因的检测。

• 发表时间: 2001
• 期刊: International Journal of Developmental Biology 45(4)
• 作者: Haruyoshi Yamaza;et al.
• 通讯作者: et al.

In Situ Expression of RANKL,RANK,OPG and Cytokines in Osteoclasts of Rat Periodontal Tissue.

大鼠牙周组织破骨细胞中RANKL、RANK、OPG和细胞因子的原位表达。

• 发表时间: 2004
• 期刊: Journal of Periodontal Research 39(1)
• 作者: T.Ogasawara;Y;et al.
• 通讯作者: et al.

Expression pattern of Epstein-Barr virus latent genes in cell lines derived from oral squamous cell carcinoma.

口腔鳞状细胞癌细胞系中 Epstein-Barr 病毒潜伏基因的表达模式。

• 发表时间: 2004
• 期刊: Oral Medicine and Pathology 9
• 作者: A.M.E.Rasul;et al.
• 通讯作者: et al.