Three-dimensional localization of a gamma-tubulin-like molecule in the enterocyte of the mouse duodemun and its molecular analysis

小鼠十二指肠肠上皮细胞中γ-微管蛋白样分子的三维定位及其分子分析

• 批准号: 14570014
• 负责人: SUZAKI Etsuko
• 金额: $ 1.22万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570014/
• 关键词: gamma-tubulin; microtubule; cell adhesion molecule; cells-meeting junction; three-dimensional analysis; proteome analysis; NOXO 1; immunoelectron microscopy; NOXO1; 3細胞ジャンクション; 免疫蛍光染色

Microtubules play important roles in maintaining the cell morphology and polarity. We had been studying to clarify the localization of gamma-tubulin in a polarized cell which organizes microtubule arrays and found that one of anti-gamma-tubulin antibody, G9, stained three-cells meeting junctions in the mouse duodenal enterocyte. Therefore, we have investigated the precise localization of G9-staining molecule and its molecular property for the two years of 2002-2003G9 staining was examined by using a confocal microscope, and it was shown that G9 stained three-cells-junction in the middle to upper part of lateral membranes. Especially, the staining of the top regions were strong but was totally different from the one of E-cadherin. Immunoelectronmicroscopy was further performed, and the labels of an anti-gamma-tubulin antibody and G9 were seen around the three-cells-meeting junctions where the labels of an anti-alfa-tubulin antibody were also detected. These results confirmed that a gamma-tubulin-like molecule localizes at three-cells-meeting junction associating with microtubulesIn order to clarify G9-recognising molecule, the proteome analysis and succeeding peptide-MASS-fingerprint analysis was also performed. The proteome analysis demonstrated that there was one candidate which would be valuable for the further analysis. The peptide-MASS-fingreprint analysis suggests that the G9-recognizing molecule is possibly NOXO (NASPH-oxidase organizer) 1 whose c-DNA was newly identified in 2003. Further investigations will be needed to work out more details

微管在维持细胞形态和极性方面起着重要作用。我们一直在研究γ-微管蛋白在组织微管阵列的极化细胞中的定位，并发现抗γ-微管蛋白抗体之一G9染色小鼠十二指肠上皮细胞中的三细胞连接处。因此，我们在2002- 2003年的两年时间里，对G9染色分子的精确定位及其分子特性进行了研究。利用共聚焦显微镜对G9染色进行了检测，结果表明，G9染色的三细胞连接位于侧膜的中上部。尤其是顶部区域的染色较强，但与E-cadherin的染色完全不同。进一步进行免疫电子显微镜检查，并且在三个细胞相遇的连接处周围观察到抗-γ-微管蛋白抗体和G9的标记，其中还检测到抗-α-微管蛋白抗体的标记。这些结果证实了γ-微管蛋白样分子定位于与微管相关的三细胞连接处。为了明确G9识别分子，还进行了蛋白质组分析和随后的肽-MASS指纹分析。蛋白质组分析表明，有一个候选，这将是有价值的进一步分析。肽-MASS-指纹分析表明，G9识别分子可能是NOXO（NASPH氧化酶组织者）1，其c-DNA是2003年新发现的。需要进一步调查以确定更多细节

项目成果

E.Suzaki, T.Suzaki, K.Kataoka: "Use of Taxol and colllagennse for better three-dimensional visualization of microtubules in the enterocyte and Brunner's gland cell, with special reference to their relation to the Golgi apparatus"Journal of Electron Micros

E.Suzaki、T.Suzaki、K.Kataoka：“使用紫杉醇和胶原蛋白更好地对肠上皮细胞和布伦纳腺细胞中的微管进行三维可视化，特别是它们与高尔基体的关系”电子显微镜杂志

A.Saito, E.Suzaki, K.Kataoka, T.Suzaki et al.: "Gliding movement in Peranema trichophorum is powered by flagellar surface motility"Cell Motility and the Cytoskeleton. 55. 224-253 (2003)

A.Saito、E.Suzaki、K.Kataoka、T.Suzaki 等人：“Peranema trichophorum 的滑行运动由鞭毛表面运动提供动力”细胞运动和细胞骨架。

A.Saito, E.Suzaki, K.Kataoka, T.Suzaki 他5名: "Gliding movement in Peranema trichophorum is powered by flagellar surface motility"Cell Motility and the Cytoskeleton. 55. 224-253 (2003)

A.Saito、E.Suzaki、K.Kataoka、T.Suzaki 和其他 5 人：“Peranema trichophorum 的滑翔运动由鞭毛表面运动提供动力”细胞运动和细胞骨架 55. 224-253 (2003)。

A.Saito, E.Suzaki, K.Kataoka, T.Suzaki 他5名: "Gliding movement in Peranema trichophorum is powered by flagellar surface motility"Cell Motility and the Cytoskeleton. (in press).

A.Saito、E.Suzaki、K.Kataoka、T.Suzaki 和其他 5 人：“Peranema trichophorum 的滑行运动由鞭毛表面运动提供动力”细胞运动和细胞骨架（正在出版）。

E.Suzaki, T.Suzaki, K.Kataoka: "Use of Taxol and collagenase for better three-dimensional visualization of microtubules in the enterocyte and Brunner's gland cell, with special reference to their relation to the Golgi apparatus"Journal of Electron Microsc

E.Suzaki、T.Suzaki、K.Kataoka：“使用紫杉醇和胶原酶更好地对肠上皮细胞和布伦纳腺细胞中的微管进行三维可视化，特别是它们与高尔基体的关系”电子显微镜杂志