Translocation of Transfected GLUT2 to the Apical Membrane in Rat Intestinal IEC-6 Cells
| Autor: | Ye Zheng, Michael G. Sarr |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
endocrine system
DNA Complementary Physiology Phloretin Glucose uptake Transfection digestive system Microtubules Article chemistry.chemical_compound Animals Cytochalasin B Protein kinase C Protein Kinase C Glucose Transporter Type 2 biology Cell Membrane Gastroenterology Glucose transporter Biological Transport Apical membrane Molecular biology Tubulin Modulators Cell biology Rats Calphostin C Enterocytes Glucose Phlorhizin chemistry Intestinal Absorption biology.protein GLUT2 Tetradecanoylphorbol Acetate Signal Transduction |
| Popis: | In this study, we transfected the full length cDNA of glucose transporter 2 (GLUT2) into IEC-6 cells (which lack GLUT2 expression) to investigate GLUT2 translocation in enterocytes. The purpose of this study was to investigate cellular mechanisms of GLUT2 translocation and its signaling pathway. Rat GLUT2 cDNA was transfected into IEC-6 cells. Glucose uptake was measured by incubating cell monolayers with glucose (0.5–50 mM), containing 14C-d-glucose and 3H-l-glucose, to measure stereospecific, carrier-mediated and passive uptake. We imaged GLUT2 immunoreactivity by confocal fluorescence microscopy. We evaluated the GLUT2 inhibitor (1 mM phloretin), SGLT1 inhibitor (0.5 mM phlorizin), disrupting microtubular integrity (2 μM nocodazole and 0.5 μM cytochalasin B), protein kinase C (PKC) inhibitors (50 nM calphostin C and 10 μM chelerythrine), and PKC activator (50 nM phorbol 12-myristate 13-acetate: PMA). In GLUT2–IEC cells, the K m (54.5 mM) increased compared with non-transfected IEC-6 cells (7.8 mM); phloretin (GLUT2 inhibitor) inhibited glucose uptake to that of non-transfected IEC-6 cells (P |
| Databáze: | OpenAIRE |
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