Y Akimoto1, L K Kreppel, H Hirano, G W Hart. 1. Department of Biochemistry and Molecular Genetics Schools of Medicine and Dentistry, University of Alabama at Birmingham Station, USA.
Abstract
AIMS/HYPOTHESIS: Streptozotocin (STZ), a chemically reactive analogue of N-acetylglucosamine, induces necrosis of the beta cells, resulting in diabetes mellitus. Glucose-induced insulin resistance is mediated by increased activity of the hexosamine pathway. We aimed to examine the regulation of O-GlcNAc transferase expression and activity in the normal and streptozotocin diabetic pancreas. METHODS: Rats were made diabetic by an injection of streptozotocin (65 mg/kg). The expression of O-GlcNAc transferase protein was examined by immunoblot analysis. Activity of O-GlcNAc transferase was assayed by the incorporation of [3H]GlcNAc into the synthetic peptide. Localization of O-GlcNAc transferase was done by immunohistochemistry. The change of O-GlcNAc modification of proteins was examined by immunoblot analysis. RESULTS: In the STZ-induced diabetic pancreas, a severe loss of beta cells was observed, whereas alpha cells had increased in number. The diabetic pancreas showed an increase in the expression of O-GlcNAc transferase at the protein level and the O-GlcNAc transferase activity in it was increased significantly (p < 0.05). An increase in the immunostaining intensity in the cytoplasm of islet beta cells was also observed in the diabetic pancreas, whereas exocrine cells and islet cells other than beta cells showed little change in immunostaining intensity. The pancreas of STZ-diabetic rats showed a 3.1-fold increase in total cellular O-GlcNAc-modified proteins. CONCLUSION/ INTERPRETATION: These findings indicate that O-GlcNAc transferase plays an important part in the modulation of O-GlcNAc concentrations in the pancreas and suggest that the increase in O-GlcNAc modification of the proteins correlates closely with diabetes.
AIMS/HYPOTHESIS: Streptozotocin (STZ), a chemically reactive analogue of N-acetylglucosamine, induces necrosis of the beta cells, resulting in diabetes mellitus. Glucose-induced insulin resistance is mediated by increased activity of the hexosamine pathway. We aimed to examine the regulation of O-GlcNAc transferase expression and activity in the normal and streptozotocindiabetic pancreas. METHODS:Rats were made diabetic by an injection of streptozotocin (65 mg/kg). The expression of O-GlcNAc transferase protein was examined by immunoblot analysis. Activity of O-GlcNAc transferase was assayed by the incorporation of [3H]GlcNAc into the synthetic peptide. Localization of O-GlcNAc transferase was done by immunohistochemistry. The change of O-GlcNAc modification of proteins was examined by immunoblot analysis. RESULTS: In the STZ-induced diabetic pancreas, a severe loss of beta cells was observed, whereas alpha cells had increased in number. The diabetic pancreas showed an increase in the expression of O-GlcNAc transferase at the protein level and the O-GlcNAc transferase activity in it was increased significantly (p < 0.05). An increase in the immunostaining intensity in the cytoplasm of islet beta cells was also observed in the diabetic pancreas, whereas exocrine cells and islet cells other than beta cells showed little change in immunostaining intensity. The pancreas of STZ-diabeticrats showed a 3.1-fold increase in total cellular O-GlcNAc-modified proteins. CONCLUSION/ INTERPRETATION: These findings indicate that O-GlcNAc transferase plays an important part in the modulation of O-GlcNAc concentrations in the pancreas and suggest that the increase in O-GlcNAc modification of the proteins correlates closely with diabetes.
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