Exhaustive analysis of gene expression dining bone regeneration

详尽分析促进骨再生的基因表达

• 批准号: 12671832
• 负责人: SHIBATA Yasuaki
• 金额: $ 2.56万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671832/
• 关键词: Bone; Regeneration; Mesenchymal stem cells; Osteocalcin; BrdU; Alkaline phosphatase

We conducted the following; experiments.1. Characterizaiion of mecenchymal stem cells during bone regenerationWe first made n round hole, 2.2 mm in a diameter, at diaphysis of femurs of growing mice To Investigate the proliferation activity of mesenchymal cells during the bone repair, we explored incorporation of BrdU. These studies demonstrated that proliferating mesenchymal cells incorporating BrdU increased 3 days after injury in our experimental system.2. Changes in gene expression during bone regenerationUsing above described experimental model, we investigated the changes in expression patterns of bone-related genes by Northern blot analysis. ALP mRNA increased on day 4 and osteocalcin mRNA increased OB day 5 after bone injury. Histochemical and immunohistochemical studies indicated similar expression patterns of these molecules.3. Above experiments suggested that mesenchymal stem cells appeared during days 4 and 5 after injury-in our experimental model. We are now exploring the genes related to mesenchymal stem cells by cDNA chips.

我们进行了以下实验。1.本实验首先在生长期小鼠股骨干上制作直径为2.2mm的圆孔，观察骨髓间充质干细胞在骨修复过程中的增殖活性，并观察BrdU掺入情况。这些研究表明，在我们的实验系统中，损伤后3天，增殖的含BrdU的间充质细胞增加.骨再生过程中基因表达的变化利用上述实验模型，我们通过北方印迹分析研究了骨相关基因表达模式的变化。骨损伤后第4天ALP mRNA升高，第5天骨钙素mRNA升高。组织化学和免疫组织化学研究表明这些分子的表达模式相似.上述实验表明，在我们的实验模型中，间充质干细胞在损伤后4天和5天出现。我们正在利用cDNA芯片探索间充质干细胞的相关基因。

项目成果

Takahashi H, Fujita S, Shibata Y, Yamaguchi A: "Adenomatoid odontogenic tumour : immunohistochmical demonstration of transferrin, ferritin and alpha-one-antitrypsin"J Oral Pathol Med. 32. 237-244 (2001)

Takahashi H、Fujita S、Shibata Y、Yamaguchi A：“腺瘤样牙源性肿瘤：转铁蛋白、铁蛋白和 α-1-抗胰蛋白酶的免疫组织化学演示”J Oral Pathol Med。

Yamamoto-Fukuda T., Shibata Y., Hishikawa Y., Shin M., Yamaguchi A., Kobayashi T., Koji T.: "Effects of various decalcification protocols on detection of DNA strand breaks by terminal dUTP nick end labelling"Histochem. J.. 32. 697-702 (2000)

Yamamoto-Fukuda T.、Shibata Y.、Hishikawa Y.、Shin M.、Yamaguchi A.、Kobayashi T.、Koji T.：“各种脱钙方案对末端 dUTP 缺口末端标记检测 DNA 链断裂的影响”Histochem

Tomomi Yamamoto: "Effects of various decalcification protocols on detection of DNA strand breaks by terminal dUTP nick end labelling"The Histochemical Journal. 32. 697-702 (2000)

Tomomi Yamamoto：“各种脱钙方案对通过末端 dUTP 缺口末端标记检测 DNA 链断裂的影响”《组织化学杂志》。

Takahashi H., Fujita S., Shibata Y., Yamaguchi A.: "Adenomiioid odontogcaic tumour immunohistochemical demonstration of transferrin, ferritin and alpha-one antitrypsin"J. Oral Pathol. Med.. 30. 237-244 (2001)

Takahashi H.、Fujita S.、Shibata Y.、Yamaguchi A.：“腺样齿状肿瘤免疫组织化学证明转铁蛋白、铁蛋白和 α-1 抗胰蛋白酶”J。

Shibata Y, Hishikawa Y et al.: "Assessment of decalcifiying protocols for detection of specific RNA by non-radioactive in situ hybridization in calcified tissues"Histochem Cell Biol. 113. 153-159 (2000)

Shibata Y、Hishikawa Y 等人：“通过钙化组织中的非放射性原位杂交检测特定 RNA 的脱钙方案的评估”Histochem Cell Biol。