BIOGENESIS OF SECRETORY GRANULES IN PAROTID ACINAR CELLS

腮腺腺泡细胞分泌颗粒的生物发生

• 批准号: 09671909
• 负责人: YOKOYAMA Miki
• 金额: $ 1.92万
• 依托单位: NIHON UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671909/
• 关键词: RAT PAROTID; SMALL-MOLECULAR-MASS GTP-BINDING PROTEINS; ACINAR CELLS; SECRETORY GRANULES; EXOCRINE GLANDS; SALIVERY GLANDS; ADP-RIBOSYLATION FACTOR; ADPリボシレーション因子

In the previous study (1997), we demonstrated that ADP-ribosylation factor (Arf) is present in the cytosolic fraction of rat parotid acinar cells and Arf translocates to the secretory granules in a GTp-dependent manner. So far, six mammalian Arf genes (Arf1-Arf6) have been reported. In bovine adrenal chromaffin cells, Arf6 is associated to the secretory granules. Now we have succeeded in demonstrating the following two points to elucidate the role of Arf proteins on the secretory granules. These results were published in Archives of Biochemistry and Biophysics 357, 147-154 (1998).1. Identification of the Arf isotype as Arfi1that translocates to the secretory granulesWe identified the Arf isotype that translocates to the secretory granules by using antibodiesspecific for Arf1, Arf3, Arf5, and Arf6. In contrast to the Arf6 localization to the secretory granules in bovine adrenal chromaffin cells, Arf6 was not detected in the granule membrane fraction of rat parotid acinar cells.2. Demonstration of the Arfi translocation to the secretory granules by immunoelectron microscopyTo confirm directly that Arf1 is translocated to the intact secretory granules, the secretorygranules were isolated after the translocation experiment and used for the immunoelectron microscopy with anti-Arf antibody. Gold particles were detected around the surface of the mature secretory granules, about 1 mum in diameter.

在以前的研究中（1997年），我们证明，ADP-核糖基化因子（Arf）是存在于大鼠腮腺腺泡细胞的胞质部分和Arf易位的分泌颗粒中的GTP依赖性的方式。目前已报道的哺乳动物Arf基因有6个（Arf 1-Arf 6）。在牛肾上腺嗜铬细胞中，Arf 6与分泌颗粒相关。现在，我们已经成功地证明了以下两点来阐明Arf蛋白在分泌颗粒上的作用。这些结果发表在Archives of Biochemistry and Biophysics 357，147-154（1998）中。Arf同种型的鉴定Arf 1转位到分泌颗粒我们通过使用Arf 1、Arf 3、Arf 5和Arf 6的特异性抗体鉴定Arf同种型转位到分泌颗粒。与牛肾上腺嗜铬细胞分泌颗粒中的Arf 6不同，大鼠腮腺腺泡细胞颗粒膜部分中未检测到Arf 6.免疫电镜显示Arf 1转位到分泌颗粒中为了直接证实Arf 1转位到完整的分泌颗粒中，在转位实验后分离分泌颗粒，并用抗Arf抗体进行免疫电镜观察。在成熟分泌颗粒的表面周围检测到金颗粒，直径约1 μ m。

项目成果

J.Fujita-Yoshigaki, Y.Dohke, M.Hara-Yokoyama, S.Furuyama and H.Sugiya: "SNARE PROTEINS ESSENTIAL FOR CYCLIC AMP-REGULATED EXOCYTOSIS IN SALIVARY GLANDS" European Journal of Morphology. 36. 46-49 (1998)

J.Fujita-Yoshigaki、Y.Dohke、M.Hara-Yokoyama、S.Furuyama 和 H.Sugiya：“唾液腺中循环 AMP 调节的胞吐作用所必需的圈套蛋白”欧洲形态学杂志。

Yoko Dohke et al.: "ADP-RIBOSYLATION FACTORS IN RATPAROTID ACINAR CELLS" European Journal of Morphology. 36. 186-189 (1998)

Yoko Dohke 等人：“大鼠腺泡细胞中的 ADP-核糖基化因子”欧洲形态学杂志。

Yoko Dohke et al.: "Translocation of Arfl to the Secretory Granules in Rat Parotid Acinar Cells" ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS. 357(1). 147-154 (1998)

Yoko Dohke 等人：“大鼠腮腺腺泡细胞中 Arfl 易位至分泌颗粒”生物化学和生物物理学档案。

Y.Dohke, M.Hara-Yokoyama, J.Fujita-Yoshigaki, R.A.Kahn, Y.Kanaho, S.Hashimoto, S.Furuyama and H.Sugiya: "Translocation of Arf1 to the Secretory Granules in Rat Parotid Acinar Cells" Archives of Biochemistry and Biophysics. 357(1). 147-154 (1998)

Y.Dohke、M.Hara-Yokoyama、J.Fujita-Yoshigaki、R.A.Kahn、Y.Kanaho、S.Hashimoto、S.Furuyama 和 H.Sugiya：“Arf1 易位至大鼠腮腺腺泡细胞中的分泌颗粒”档案

Hiromi Michikawa et al.: "cGMP production is coupled to Ca^<2+>-dependent nitric oxide generation in rabbit parotid acinar cells"Cell Calcium. 23(6). 405-412 (1998)

Hiromi Michikawa等人：“cGMP的产生与兔腮腺腺泡细胞中Ca 2+ 依赖性一氧化氮的产生相结合”细胞钙。