1997•Cell Biochemistry and FunctionRequires access

Non-functional role of syntaxin 2 in insulin exocytosis by pancreatic β cells

Shinya Nagamatsu, Hiroki Sawa, Yoko Nakamichi, Yoshinori Kondo, Satsuki Matsushima, Takashi Watanabe

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Abstract

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.

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What this paper is about

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.

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Available abstract

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.

Key concepts: Syntaxin 3, Exocytosis, Syntaxin, Monoclonal antibody, Western blot, Cell biology, Insulin, Islet

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