MECHANISM OF EXOCYTOTIC MUCIN RELEASEIN SUBLINGUAL GLAND

舌下腺胞吐性粘蛋白释放机制

• 批准号: 09671906
• 负责人: SUGIYA Hiroshi
• 金额: $ 1.92万
• 依托单位: NIHON UNIVERSITY SCHOOL OF DENTISTRY AT MATSUDO
• 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-09671906/
• 关键词: SALIVARY GLANDS; EXOCYTOSIS; CALCIUM; SUBLINGUAL GLAND; CYCLIC GMP; NITRIC OXIDE; ARF; SYNTAXIN 1A; シンタキシン; シンコリン; 開口分泌; ムチン; 分泌顆粒

In salivary gland acinar cells, the secretory function is rcgulated by two main signaling pathways via muscarinic and beta-adrenergic receptors.Stimulation of muscarinic receptors causes the increase in intracellular calcium ions (Ca^<2+>). However, the mechanism with Ca^<2+>-dependent secretory pathway has not well elucidated. We here studied with the mechanism of exocytosis via Ca^<2+> signaling pathway.During this study, we first demonstrated that the increase in intracellular Ca^<2+> induced by muscarinic receptor stimulation activated nitric oxide synthase. Subsequently, nitric oxide generated stimulated soluble guanylyl cyclase and induced cyclic GMP formation in salivary gland acinar cells. We next found that Arfi, a small GTP-binding protein, existed in rat parotid acinar cells. Furthermore, we demonstrated that the Arf1 translocated to secretory granules in the presence of GTP gammaS.Rat sublingual gland acinar cells are an useful model for the study with Ca^<2+>-dependent exocytosis, since Ca^<2+>-mobilizing receptor activation induces mucin exocytosis. We found that high concentration of syntaxin1A in the membrane fraction of sublingual acinar cells by immuno blotting.The expression of mRNA of syntaxin1A was detected in rat salivary glands such as sublingual and submandibular glands. However, we failed to detectthe binding proteins to syntaxinlA such as VAMP-2 and SNAP-25.Further studies with the relationship between syntaxin1A and Ca^<2+>-nitric oxide-cyclic GMP signaling and identification of syntaxin 1A binding proteins are necessary.

在唾液腺腺泡细胞中，分泌功能由通过毒蕈碱受体和β-肾上腺素能受体的两条主要信号传导途径调节。刺激毒蕈碱受体导致细胞内钙离子(Ca^2+)增加。然而，Ca^2+依赖性分泌途径的机制尚未得到很好的阐明。我们在此研究了Ca^2+信号通路的胞吐作用机制。在本研究中，我们首先证明了毒蕈碱受体刺激诱导的细胞内Ca^2+增加激活了一氧化氮合酶。随后，一氧化氮刺激可溶性鸟苷酸环化酶并诱导唾液腺腺泡细胞中环鸟苷酸的形成。接下来我们发现 Arfi，一种小的 GTP 结合蛋白，存在于大鼠腮腺腺泡细胞中。此外，我们证明，在 GTP gammaS 存在的情况下，Arf1 易位至分泌颗粒。大鼠舌下腺腺泡细胞是研究 Ca^<2+> 依赖性胞吐作用的有用模型，因为 Ca^<2+> 动员受体激活诱导粘蛋白胞吐作用。通过免疫印迹发现舌下腺泡细胞膜部分存在高浓度的syntaxin1A。在大鼠舌下腺、颌下腺等唾液腺中检测到syntaxin1A mRNA的表达。然而，我们未能检测到syntaxin1A的结合蛋白，如VAMP-2和SNAP-25。需要进一步研究syntaxin1A与Ca^2+-一氧化氮-环GMP信号传导之间的关系以及syntaxin 1A结合蛋白的鉴定。 。

项目成果

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

Michikawa H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有关。”

Dohke Y,Hara-Yokoyama M,Fujita-Yoshigaki J,Furuyama S & Sugiya H: "ADP-ribosylation factors in rat parotid acinar cells." European J Morphology. 36. 186-189 (1998)

Dohke Y、原横山 M、藤田吉垣 J、古山 S

Sugiya H,Michikawa H,Mitsui Y,Fujita-Yoshigaki J,hara-Yokoyama M & Furuyama S: "Ca^<2+>-nitoric oxide- ^CGMP signaling in rabbit parotid acinar cells." European J Morphology. 36. 194-197 (1998)

杉谷 H、道川 H、三井 Y、藤田吉垣 J、原横山 M

Mitsui Y, et al.: "Nitric oxide synthase activities in mammalian parotid and submandibular salivary glands." Arch Oral Biol. 42. 621-624 (1997)

Mitsui Y 等人：“哺乳动物腮腺和下颌下唾液腺中的一氧化氮合酶活性。”

Fujita-Yoshigaki J,Dohke Y,Hara-Yokoyama M,Furuyama S & Sugiya H: "SNARE proteins essential for Cyclic AMP-regulated exocytosis in salivary glands." European J Morphology. 36. 46-49 (1998)

藤田吉垣 J、Dohke Y、原横山 M、古山 S