Nagamatsu S, Sawa H, Nakamichi Y, Kondo Y, Matsushima S, Watanabe T
Department of Biochemistry, Kyroin University School of Medicine, Tokyo, Japan.
Cell Biochem Funct. 1997 Dec;15(4):237-42. doi: 10.1002/(SICI)1099-0844(199712)15:4<237::AID-CBF746>3.0.CO;2-U.
This study was designed in order to examine the expression and functional role of syntaxin 2/epimorphin in pancreatic beta cells. Northern blot analysis revealed that syntaxin 2 mRNA was able to be detected in mouse beta TC3 cells, but not in isolated mouse islets. In agreement with this result, immunoblot analysis detected an appreciable amount of syntaxin 2 protein in beta TC3 cells, but not in mouse islets. Immunohistochemistry of the mouse pancreas demonstrated that syntaxin 2 was little evident in islet cells of Langerhans, and somewhat predominant in exocrine tissues. In order to examine whether syntaxin 2 is anchored to cell surfaces in beta TC3 cells, living cells were incubated with a monoclonal antibody against syntaxin 2 (MC-1). The antibody bound to their surfaces, indicating that syntaxin 2 was localized on cell surfaces. The addition of MC-1 to the culture medium of beta TC3 cells did not affect insulin release under the presence or absence of 11 mM glucose, indicating that syntaxin 2 is not associated with insulin exocytosis. Thus, the expression of syntaxin 2 in islets of Langerhans is very low and the function of this protein is probably unrelated to the insulin exocytosis pathway.
本研究旨在检测 syntaxin 2/上皮形态发生素在胰腺β细胞中的表达及功能作用。Northern 印迹分析显示,在小鼠βTC3 细胞中能检测到 syntaxin 2 mRNA,但在分离的小鼠胰岛中未检测到。与此结果一致,免疫印迹分析在βTC3 细胞中检测到相当数量的 syntaxin 2 蛋白,但在小鼠胰岛中未检测到。小鼠胰腺的免疫组织化学显示,syntaxin 2 在胰岛β细胞中不太明显,而在外分泌组织中较为突出。为了检测 syntaxin 2 在βTC3 细胞中是否锚定在细胞表面,将活细胞与抗 syntaxin 2 单克隆抗体(MC-1)孵育。该抗体与细胞表面结合,表明 syntaxin 2 定位于细胞表面。在有或无 11 mM 葡萄糖存在的情况下,向βTC3 细胞培养基中添加 MC-1 均不影响胰岛素释放,表明 syntaxin 2 与胰岛素胞吐作用无关。因此,syntaxin 2 在胰岛中的表达非常低,且该蛋白的功能可能与胰岛素胞吐途径无关。