Direct-3D-observation of intracellular organella

细胞内细胞器的直接 3D 观察

• 批准号: 10670017
• 负责人: SUZAKI Etsuko
• 金额: $ 2.11万
• 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: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670017/
• 关键词: Direct-3D-observation; Microtubules; γ-tubulin; Duodenum; Parietal Cells; Gastric acid secretion; Stomach; Three-dimensional Structure; ゴルジ装置; レクチン細胞化学; 吸収上皮細胞; 十二指腸腺細胞; 胃粘膜; 十二指腸腺

Owing to this grant, we were able to perform three-dimensional observation of intracellular organella by using a direct-3D-microscope equipped with a double dichroic mirror, which enabled us to obtain 2-colored 3D images under the microscopic field. The main results during this research period were as below.1. The direct-3D-microscope was found to be a simple and useful for three-dimensional examination of tissue sections. This will be the most economical equipment for the 3D observation.2. Successful staining of microtubules in epithelial cells of the mouse duodenum was accomplished, and the spatial relationship between the Golgi apparatus and its supporting element of microtubules was clarified. Microtubules have been known to form an intracellular array radiating from a core called microtubule organizing center (MTOC). In order to clarify the localization of MTOC, a MTOC-associating molecule, gamma-tubulin, was further examined immunocytochemically.3. The intracellular components of a parietal cell in the rat stomach were stained with anti-proton-pump antibodies. The changes in the staining patterns were suggested to be related to the gastric acid secretion. The double staining with proton-pump and mitochondria showed the efficient localization for energy-dependent secretion of the gastric acid.

由于这项资助，我们能够通过使用配备双分色镜的直接3D显微镜对细胞内细胞器进行三维观察，这使我们能够在显微镜下获得2色3D图像。主要研究结果如下：1.直接三维显微镜被认为是一种简单而有用的三维检查组织切片。这将是最经济的三维观测设备。2.成功地对小鼠十二指肠上皮细胞中的微管进行了染色，并阐明了高尔基体与其微管支持元件之间的空间关系。已知微管形成从称为微管组织中心（MTOC）的核心辐射的细胞内阵列。为了明确MTOC的定位，进一步用免疫细胞化学方法检测了MTOC相关分子γ-微管蛋白.用抗质子泵抗体对大鼠胃壁细胞的细胞内成分进行染色。结果表明，胃粘膜上皮细胞的染色模式改变与胃酸分泌有关。质子泵和线粒体的双重染色显示了能量依赖性胃酸分泌的有效定位。

项目成果

Takafumi Ito: "Effect of glucose on intercellular junctions of cultured human peritoneal mesothelial cells"J.Am.Soc.Nephrol.. 11. 1969-1979 (2000)

Takafumi Ito：“葡萄糖对培养的人腹膜间皮细胞细胞间连接的影响”J.Am.Soc.Nephrol.. 11. 1969-1979 (2000)

Etsuko Suzaki: "Three-dimensional visualization of the Golgi apparatus and its supporting elements of microtubules in a tissue section"Bioimaging. 9(in Japanese). 140-141 (2000)

Etsuko Suzaki：“组织切片中高尔基体及其微管支撑元件的三维可视化”生物成像。

洲崎悦子: "ゴルジ装置の三次元構造"バイオイメージング. 7. 179-180 (1998)

Etsuko Susaki：“高尔基体的三维结构”生物成像 7. 179-180 (1998)。

片岡勝子: "細胞更新の超微形態:胃粘膜の表層粘液細胞"細胞. 32. 289-293 (2000)

Katsuko Kataoka：“细胞更新的超形态学：胃粘膜的表面粘液细胞”细胞。 32. 289-293 (2000)

"The tissue formation and intercellular connection in the gastric mucosa"Biomechanism gakkaisi. 23(in Japanese). 199-202 (1999)

“胃粘膜的组织形成和细胞间连接”生物机制gakkaisi。