Localization of the O-linked N-acetylglucosamine transferase in rat pancreas

Yoshihiro Akimoto, Lisa K. Kreppel, Hiroshii Hirano, Gerald Warren Hart

Research output: Contribution to journalArticlepeer-review

Abstract

O-linked N-acetylglucosamine transferase (OGT) catalyzes the attachment of N-acetylglucosamine (GlcNAc) monosaccharides to the hydroxyl group of serine or threonine residues of intracellular proteins and may play an important role in the hexosamine pathway. Glucose-induced insulin resistance is mediated by increased activity of the hexosamine pathway. In the present study, we examined the localization of OGT mRNA and OGT protein in the rat pancreas. The sites of OGT mRNA expression were determined by in situ hybridization histochemistry with a digoxigenin (DIG)-labeled antisense cRNA probe. Intense hybridization signals were present in the exocrine acinar cells, while weaker ones were detected in the islets of Langerhans. This distribution was confirmed using additional antisense cRNA or oligo-cDNA probes complementary to different regions of OGT mRNA. In addition, immunofluorescence staining with antibody raised against OGT stained both the exocrine acinar cells and endocrine islet cells. In the acinar cell nucleus, the zymogen granule region and contour of the cell were intensely stained. In the islets of Langerhans, especially in the α-cells, intense staining with anti-OGT antibody was observed. These staining patterns were almost identical to those seen when staining for the O-linked GlcNAc (O-GlcNAc) modification. Immuno-electron microscopy showed that OGT is localized to the euchromatin of the nucleus and around the secretory granules of exocrine acinar cells and endocrine islet cells. These results suggest that OGT is involved in the regulation of transcription and of granular secretion. Thus, one or more O- GlcNAcylated proteins may be important components of the glucose-sensing mechanism in the pancreas.

Original languageEnglish (US)
Pages (from-to)2407-2413
Number of pages7
JournalDiabetes
Volume48
Issue number12
StatePublished - Dec 1999

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism

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