Immunocytochemical dissection of Golgi apparatus.

高尔基体的免疫细胞化学解剖。

• 批准号: 08670032
• 负责人: TAMAKI Hideaki
• 金额: $ 0.9万
• 依托单位: KITASATO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08670032/
• 关键词: Golgi apparatus; Immunocytochemistry; Ultrathin cryosection; Parotid gland; beta-COP; Mannosidase II; Brefeldin A; Okadaic acid; 外分泌

To study membrane dynamics of the Golgi apparatus, we examined distributional changes of Golgi-associated proteins in rat parotid acinar cells after the treatment with Golgi-disrupting agents by immunoflurorescent microscopy and immunogold labeling on ultrathin cryosections.In untreated acinar cells, immunoreactivity of beta-COP was mainly associated with the region of Golgi apparatuson which 58K and man II were co-localized. BFA caused rapid redistribution of 58K and man II into the endoplasmic reticulum (ER) accompanied by marked vesiculation of the Golgi stack. beta-COP lost its association with the Golgi region and was dispersed throughout the cytoplasm. OA also induced rapid and marked vesiculation of the stacked structure of the Golgi apparatus, but the redistribution of 58K and man II into the ER could not be observed. Of interest is that beta-COP was markedly accumulated in the region of the disrupting Golgi apparatus on which 58K and man II immunoreactivies were preserved. Detailed observation on ultrathin crysections of OA-treated cells indicated that the Golgi cluster was composed of beta-COP-positive and-negative tubulovesicles.These results suggested that BFA inhibited attachment of beta-COP to membranes, resulting in the disturbance of transport vesicle formation. The interruption of pre-and intra-Golgi transport caused redistribution of Golgi membrane proteins follwed by disorganization of the stacked structure. OA also induced disorganization of the stack, but the mechanism differed from that of BFA.OA seemed to inhibit the fusion of transport vesicles with the target membranes but not to prevent their formation. As a result, beta-COP-coated membranes failed to be integrated into the Golgi apparatus and accumulated in the Golgi region. Normal transport is quite essential for the maintenance of the stacked structure of the Golgi apparatus.

为了研究高尔基体的膜动力学，我们用免疫荧光显微镜和免疫胶体金标记技术观察了高尔基体破坏剂处理后大鼠腮腺腺泡细胞中高尔基体相关蛋白的分布变化，在未处理的腺泡细胞中，β-COP的免疫反应主要与高尔基体的58 K和man II共定位的区域有关。BFA引起快速重新分配的58 K和曼II到内质网（ER）伴随着显着的囊泡的高尔基体堆栈。β-COP失去了与高尔基体区域的结合，并分散在整个细胞质中。OA也诱导高尔基体的堆叠结构迅速和显着的泡状化，但58 K和man II重新分配到ER中不能观察到。有趣的是，β-COP显着积累在该地区的破坏高尔基体上的58 K和man II免疫反应被保存。对OA处理的细胞的电镜观察表明，高尔基体簇由β-COP阳性和阴性的管状囊泡组成，提示BFA抑制了β-COP与细胞膜的结合，从而干扰了运输囊泡的形成。前和内高尔基体运输的中断导致高尔基体膜蛋白的重新分布，随后是堆叠结构的解体。OA也能诱导细胞膜堆叠的解体，但其机制与BFA不同，OA似乎能抑制转运囊泡与靶膜的融合，但不能阻止它们的形成。结果，β-COP包被的膜未能整合到高尔基体中并在高尔基体区域中积累。正常的运输对于维持高尔基体的堆叠结构是非常必要的。

项目成果

Hideaki Tamaki: "Three-dimensional Dynamics of the Golgi Apparatus in Mitotic Parotid Acinar Cells" Acta Histochem.Cytochem.30(5&6)(in press). (1997)

Hideaki Tamaki：“有丝分裂腮腺腺泡细胞中高尔基体的三维动力学” Acta Histochem.Cytochem.30(5

Hideaki Tamaki: "Three-dimensional Dynamics of the Golgi Apparatus in Mitotic Parotid Acinar Cells : Computer-aided Reconstruction from Cytochemically-marked Ultrathin Serial Sections" Acta Histochem.Cytochem.30(5&6). 643-651 (1997)

Hideaki Tamaki：“有丝分裂腮腺腺泡细胞中高尔基体的三维动力学：从细胞化学标记的超薄连续切片进行计算机辅助重建” Acta Histochem.Cytochem.30(5

Shohei Yamashina: "The Significance of Lamellated Structure of Golgi Apparatus in Parotid Acinar Cells." European Journal of Morphology. 34(3). 229-232 (1996)

Shohei Yamashina：“腮腺腺泡细胞中高尔基体层状结构的意义”。

Shohei Yamashina: "The Significance of Lamellated Structure of Golgi Apparatus in Parotid Acinar Cells." Eur.J.Morph.34(3). 229-232 (1996)

Shohei Yamashina：“腮腺腺泡细胞中高尔基体层状结构的意义”。

Hideaki Tamaki: "The trans Golgi Network : Its History, Structure and Function. (Japanese)" Electron Microscopy. 32(3). 137-143 (1997)

Hideaki Tamaki：“反式高尔基体网络：它的历史、结构和功能。（日语）”电子显微镜。