Molecular Mechanism and Signal Transduction for Salivary Secretion

唾液分泌的分子机制和信号转导

• 批准号: 07407051
• 负责人: SHIMONO Masaki
• 金额: $ 17.98万
• 依托单位: TOKYO DENTAL COLLEGE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07407051/
• 关键词: molecular mechanism; signal transduction; salivary secretion; connexins; endocytosis; gap junction; tight junction; immunohistocytochemistry; 唾液分泌; 腺房細胞; connexin; 免疫組織化学

The purpose of this study is to investigate the molecular mechanism of salivary secretion, which is indicated by serial phenomena including synthesis of salivary protein, sorting, intracellular transport of secretory granules and membrane fusion at the exocytosis through the signal transduction. Gap junctions which possess a signal function for salivary secretion are composed of proteins termed as connexins. It is well known that different types of connexins exist in various tissues and organs. Using western blotting, immunohistochemistry and immunocytochemistry, we demonstrated that both connexins 26 and 32 were Iocated between acinar cells, but connexin 43 was distributed between myoepithelial cells in the salivary glands. These may suggest that both connexins 26 and 32 participate in signal transduction for salivary secretion in acinar cells, but connexin 43 is correlated to signaling for contraction of myoepithelial cells (J Histochem Cytochem 44 : 49-56,1996 ; Europ J Morphol 34 : … More 197-202,1996). We also clarified that connexins 26 and 32 constituted the same gap junction in the salivary gland (Acta Histochem Cytochem, in press). The expression of connexins 32 and 43 was investigated in the developing salivary glands employing PCR,in situ hybridization and immunohistochemistry (Europ J Morphol, in press). We studied not only the localization of actin filaments and proteins constituting tight junction such as ZO-1 or occludin but also the expression of clathlin participating endocytosis and a membrane-fusion associated protein (synaptophysin) in acinar cells during secretion using immunohistochemistry, immunocytochemistry and confocal laser scanning microscope. There was a close relationship between the localization of actin filaments and ZO-1 or Occludin. Clathrin was located at the periphery of the intracellular canalicula 10 or 30 minutes after IPR stimulation. This may indicate that endocytosis of secretory-granule-membrane occur immediately after the secretion (manuscript, in preparation). Less

本研究的目的是探讨唾液分泌的分子机制，包括唾液蛋白的合成、分泌颗粒的分选、分泌颗粒的细胞内运输以及通过信号转导在胞吐时的膜融合。缝隙连接具有唾液分泌的信号功能，由连接蛋白组成。众所周知，不同类型的连接蛋白存在于各种组织和器官中。用免疫印迹、免疫组织化学和免疫细胞化学方法证明，在涎腺中，连接蛋白26和32均位于腺泡细胞之间，而连接蛋白43则分布于肌上皮细胞之间。这些可能提示连接蛋白26和32都参与腺泡细胞唾液分泌的信号转导，但连接蛋白43与肌上皮细胞收缩信号有关(组织化学细胞化学44：49-56,1996；Europ J Morphol 34：…更多197-202,1996)。我们还澄清了连接蛋白26和32在唾液腺中构成了相同的缝隙连接(印刷中的组织化学学报)。应用聚合酶链式反应、原位杂交和免疫组织化学方法(Europ J Morphol，PRESS)对发育中的涎腺组织中连接蛋白32和43的表达进行了研究。我们用免疫组织化学、免疫细胞化学和激光共聚焦扫描显微镜研究了腺泡细胞分泌过程中肌动蛋白细丝和紧密连接蛋白如ZO-1或occludin的定位，以及参与胞吞作用的胞膜蛋白和膜融合相关蛋白(突触素)在腺泡细胞中的表达。肌动蛋白细丝的定位与ZO-1或occludin密切相关。IPR刺激10min或30min后，胞内小管周围可见网状蛋白。这可能表明分泌颗粒膜的内吞作用发生在分泌物(手稿，正在准备中)之后。较少

项目成果

Minaguchi, K.et.al.: "Sac I restriction fragment length polymorphism (RFLP) related to the human CST2 gene." (in contribution).

Minaguchi, K.et.al.：“与人类 CST2 基因相关的 Sac I 限制性片段长度多态性 (RFLP)。”

Shintani, M.et.al.: "Genetic polymorphism of salivary cystatin detected by acidic polyacrylamide gel electrophoresis." (in contribution).

Shintani, M.et.al.：“通过酸性聚丙烯酰胺凝胶电泳检测唾液胱抑素的基因多态性。”

Shimono, M.et.al.: "Connexins in the developing salivary glands." Europ J Morphol. (in press). (1998)

Shimono, M.et.al.：“发育中的唾液腺中的连接蛋白。”

水口 清: "唾液成分による生体の防御" 歯界展望. 86. 650-661 (1995)

Kiyoshi Mizuguchi：“唾液成分的身体防御”Dental Perspective 86. 650-661 (1995)。

Kiyoshi Minaguchi, E Saitoh, S.Isemura and K.Sanada.: "Sac I restriction fragment length polymorphism (RFLP) related to the human CST2 gene." (in contribution).

Kiyoshi Minaguchi、E Saitoh、S.Isemura 和 K.Sanada.：“与人类 CST2 基因相关的 Sac I 限制性片段长度多态性 (RFLP)。”